Enhancing levan biosynthesis by destroying the strongly acidic environment caused by membrane-bound glucose dehydrogenase (mGDH) in Gluconobacter sp. MP2116.

Levan produced by Gluconobacter spp. has great potential in biotechnological applications. However, Gluconobacter spp. can synthesize organic acids during fermentation, resulting in environmental acidification. Few studies have focused on the effects of environmental acidification on levan synthesis. This study revealed that the organic acids, mainly gluconic acid (GA) and 2-keto-gluconic acid (2KGA) secreted by Gluconobacter sp. MP2116 created a highly acidic environment (pH < 3) that inhibited levan biosynthesis. The levansucrase derived from strain MP2116 had high enzyme activity at pH 4.0 âˆ¼ pH 6.5. When the ambient pH was less than 3, the enzyme activity decreased by 67 %. Knocking out the mgdh gene of membrane-bound glucose dehydrogenase (mGDH) in the GA and 2KGA synthesis pathway in strain MP2116 eliminated the inhibitory effect of high acid levels on levansucrase function. As a result, the levan yield increased from 7.4 g/l (wild-type) to 18.8 g/l (Δmgdh) during fermentation without pH control. This study provides a new strategy for improving levan production by preventing the inhibition of polysaccharide synthesis by environmental acidification.

Impact of preacidification on the functionality and insoluble calcium levels of Low-Moisture Part-Skim Mozzarella made from high-casein milk.

The use of low-concentration factor ultrafiltered (LCF-UF) milk for cheesemaking has become popular in recent years. Research on using LCF-UF milk to make low-moisture part-skim (LMPS) Mozzarella manufacture is limited due to concerns about the negative impact of the higher casein content on functional properties, such as melt and stretch. Pre-acidification (PA) (reflects acid added to milk before the addition of the starter culture) has been used in low-fat and nonfat Mozzarella cheeses to lower calcium levels and improve their texture and melting properties. We explored the use of PA to reduce the insoluble calcium content of cheese (calcium that is a crosslinking material for the casein proteins) and its impact on the functionality of LMPS Mozzarella made from high casein (4.2%) milk. Seven vats of LMPS Mozzarella, with a control (no PA), 2 pH values for PA (6.40 and 6.00) and 3 different acidification methods (use of acetic, citric, or carbonic acid from carbon dioxide) were evaluated for their impact on solubilization of colloidal calcium phosphate during LMPS manufacture. Composition, proteolysis, microstructural and functional changes in the LMPS cheeses were monitored during 90 d of refrigerated storage. All cheeses had similar composition except for the cheese made with milk PA with carbonic acid, which had a lower fat-in-dry matter probably due to foaming observed in this sample during the gas sparging. The cheese made with milk PA with citric acid to pH 6.00 had the lowest amount of total and insoluble calcium compared with other cheeses at all steps during cheese manufacture and storage, likely due to the calcium chelating ability of citric acid. The cheese that was made with milk PA with citric acid to pH 6.40 had a trend of slightly lower (not significant) total and insoluble calcium during cheese manufacture compared with other cheeses that were made with milk PA to pH 6.40. The pH value in all the cheeses remained similar throughout 90 d of storage, whereas the cheese that was made with milk PA with citric acid to pH 6.00 had the lowest pH values during storage. Treatment significantly (P < 0.05) impacted instrumental hardness, the maximum loss tangent during heating, the melting temperature and the storage modulus values measured at 70°C. Proteolysis was not impacted by treatment suggesting that typical aging should occur. Cheese that was PA with citric acid to pH 6.00 initially had lower maximum loss tangent values and higher hardness, possibly due to the lower pH value of this cheese. The performance of cheeses on pizza was assessed using Sensory Spectrum® method and quantitative descriptive analysis. The PA of cheese milk with citric acid helped to reduce the insoluble Ca level and at 30 and 90 d of storage, these cheeses had lower first chew hardness, lower chewiness, and lower strand thickness values compared with other cheeses. The use of PA could help improve the functionality of LMPS Mozzarella made from milk high in casein with citric acid being the most efficient in dissolving the insoluble calcium from casein.

Quantifying the combined impacts of anthropogenic CO2 emissions and watershed alteration on estuary acidification at biologically-relevant time scales: a case study from Tillamook Bay, OR, USA.

The impacts of ocean acidification (OA) on coastal water quality have been subject to intensive research in the past decade, but how emissions-driven OA combines with human modifications of coastal river inputs to affect estuarine acidification dynamics is less well understood. This study presents a methodology for quantifying the synergistic water quality impacts of OA and riverine acidification on biologically-relevant timescales through a case study from a small, temperate estuary influenced by coastal upwelling and watershed development. We characterized the dynamics and drivers of carbonate chemistry in Tillamook Bay, OR (USA), along with its coastal ocean and riverine end-members, through a series of synoptic samplings and continuous water quality monitoring from July 2017 to July 2018. Synoptic river sampling showed acidification and increased CO 2 content in areas with higher proportions of watershed anthropogenic land use. We propagated the impacts of 1). the observed riverine acidification, and 2). modeled OA changes to incoming coastal ocean waters across the full estuarine salinity spectrum and quantified changes in estuarine carbonate chemistry at a 15-minute temporal resolution. The largest magnitude of acidification (-1.4 pH ⊤ units) was found in oligo- and mesohaline portions of the estuary due to the poor buffering characteristics of these waters, and was primarily driven by acidified riverine inputs. Despite this, emissions-driven OA is responsible for over 94% of anthropogenic carbon loading to Tillamook Bay and the dominant driver of acidification across most of the estuary due to its large tidal prism and relatively small river discharges. This dominance of ocean-sourced anthropogenic carbon challenges the efficacy of local management actions to ameliorate estuarine acidification impacts. Despite the relatively large acidification effects experienced in Tillamook Bay (-0.16 to -0.23 p H ⊤ units) as compared with typical open ocean change (approximately -0.1 pH ⊤ units), observations of estuarine pH ⊤ would meet existing state standards for pH ⊤ . Our analytical framework addresses pressing needs for water quality assessment and coastal resilience strategies to differentiate the impacts of anthropogenic acidification from natural variability in dynamic estuarine systems.

The heat is on: sensitivity of goldsinny wrasse to global climate change.

Unsustainable harvesting practices have drastically reduced fish populations globally and developments in aquaculture have increased. Unexpectedly, Atlantic salmon farming caused the opening of a new fishery in northern European countries, where previously unharvested mesopredatory species, like the goldsinny wrasse (Ctenolabrus rupestris), are captured for use as cleaner fish in pens along the coast and fjords. The goldsinny wrasse is widespread in coastal areas where it plays an ecologically important role as a predator of small invertebrates. Since climate change effects are particularly pronounced in coastal waters, it becomes urgent to understand how fish like the goldsinny will respond to global climate change, including the increasing frequency and intensity of marine heatwaves (MHWs), ocean freshening (OF) and ocean acidification (OA). To address this, we conducted a multi-stressor experiment exposing adult goldsinny to each stressor individually, as well as to all three combined. The results indicated that the goldsinny is highly affected by MHWs and extremely sensitive to a multi-stressor environment, with 34% and 53% mortality, respectively. Additionally, exposure to a MHW event, OF and multi-stressor conditions affected fish metabolism, with the highest standard metabolic- and maximum metabolic-oxygen consumption rates observed for the MHW treatment. Increases in oxidized glutathione (GSSG) and percent oxidized glutathione (% GSSG) in the livers, indicative of oxidative stress, were also seen in the MHW, OF and multi-stressor treatments. As a single stressor, OA showed no significant impacts on the measured parameters. This information is important for conservation of coastal marine environments, given the species' important role in shallow-water habitats and for management of goldsinny or other mesopredatory fish harvested in coastal ecosystems. The sensitivity of the goldsinny wrasse to future stressors is of concern, and any potential reductions in abundance as a result of climate change may lead to cascade effects with ecosystem-wide consequences.

Could future ocean acidification be affecting the energy budgets of marine fish?

With the unprecedented environmental changes caused by climate change including ocean acidification, it has become crucial to understand the responses and adaptive capacity of fish to better predict directional changes in the ecological landscape of the future. We conducted a systematic literature review to examine if simulated ocean acidification (sOA) could influence growth and reproduction in fish within the dynamic energy budget theory framework. As such, we chose to examine metabolic rate, locomotion, food assimilation and growth in early life stages (i.e. larvae and juvenile) and adults. Our goal was to evaluate if acclimatization to sOA has any directional changes in these traits and to explore potential implications for energetic trade-offs in these for growth and reproduction. We found that sOA had negligible effects on energetic expenditure for maintenance and aerobic metabolism due to the robust physiological capacity regulating acid-base and ion perturbations but substantive effects on locomotion, food assimilation and growth. We demonstrated evidence that sOA significantly reduced growth performance of fish in early life stages, which may have resulted from reduced food intake and digestion efficiency. Also, our results showed that sOA may enhance reproduction with increased numbers of offspring although this may come at the cost of altered reproductive behaviours or offspring fitness. While these results indicate evidence for changes in energy budgets because of physiological acclimatization to sOA, the heterogeneity of results in the literature suggests that physiological and neural mechanisms need to be clearly elucidated in future studies. Lastly, most studies on sOA have been conducted on early life stages, which necessitates that more studies should be conducted on adults to understand reproductive success and thus better predict cohort and population dynamics under ongoing climate change.

Mn-CDF family genes enhance the manganese tolerance of the tea plants (Camellia sinensis) under acidic condition.

The tea plants cultivated in acidic soils are vulnerable to excessive manganese (Mn), which increases the risk of Mn2+ toxicity to physiology and development. Mn-cation diffusion facilitator (CDF) family genes have been implicated in regulating Mn homeostasis and tolerance. However, the mechanism of Mn tolerance of tea plants in acidic environments is still unknown. In this study, we initially examined the phenotypic characteristics and Mn contents variability in different tissues of tea plants under various Mn concentration at pH 5 and 4. We observed that tea plants exhibited remarkably high Mn tolerance at pH 4, with Mn accumulation notably elevated in the aboveground tissues under pH 4 condition after 28-day treatment. We found the expression levels of Mn-CDF genes, had different subcellular localization, were tissue-specific and significantly induced by high Mn concentrations at pH 4 condition. Furthermore, the yeast complementation assays indicated that the heterologous expression of Mn-CDF genes restored the growth of a Mn2+ sensitive yeast strain, Δpmr1. Taken together, these results suggest that Mn-CDF family genes function as Mn transporters to participate in Mn tolerance in acidic environments. This study provides reference for further study on the mechanism of maintaining Mn homeostasis in tea plants under soil acidification.

Morphological, physiological, and transcriptomic analyses indicate that cell wall properties and antioxidant processes are potential targets for improving the aluminium tolerance of broad beans.

Aluminium (Al) stress is the second-leading abiotic stress on crops. An improved understanding of the response mechanisms of plants to Al stress will provide scientific guidance for enhancing the crops' tolerance to Al stress. In this study, Al stress (50-200 µM AlCl3) caused visible damage to broad bean (Vicia faba L.) roots rather than shoots, which was attributed to Al accumulation and distribution in different tissues. Root transcriptomic analysis revealed that Al stress altered cell wall properties by downregulating lignin synthesis and several xyloglucan endotransglucosylase/hydrolase-, expansin- and peroxidase (POD)-encoding genes, which likely weakened cell extensibility to inhibit root growth. Additionally, Al stress impeded reactive oxygen species scavenging pathways involving POD activity and flavonoid biosynthesis, leading to oxidative damage characterised by malondialdehyde accumulation. These results indicate that optimising cell wall properties and/or enhancing antioxidant processes are crucial for alleviating Al toxicity to broad beans. Interestingly, exogenous application (500 and 1000 µM) of the flavonoid apigenin effectively alleviated Al toxicity in broad bean roots by partially improving the total antioxidant capacity of the roots. This study contributes to understanding the interaction between plants and Al and provides new strategies to alleviate Al toxicity in crops.

Effect of acidification pretreatment on two-phase anaerobic digestion of acidified food waste.

Acidified food waste significantly disrupts anaerobic digestion, highlighting the need for effective solutions to mitigate its impact. This study presents a method that utilizes acidified sludge to pretreat acidified food waste, thereby significantly improving the efficiency of hydrolysis and acidogenesis. After acidification pretreatment, hydrolysis efficiency improved from 64.54 % to 96.51 %, while acidogenesis efficiency increased from 34.82 % to 49.95 %. Additionally, the concentration of short-chain fatty acids and hydrogen production in the acidification pretreatment group increased by 45.89 % and 48.67 %, respectively. The pretreatment group exhibited a biochemical methane potential of 512.84 ± 13.73 mL/(g volatile suspended solids), which was 35.77 % higher than that of the control group. Mechanism analysis revealed that the higher abundance of genes associated with lactate dehydrogenase in the acidified sludge facilitated the rapid degradation of lactic acid. Moreover, the abundant Clostridium butyricum in the acidified sludge promoted the targeted conversion of lactic acid and other organic matter into butyric acid within the food waste system. This efficient butyric acid fermentation improved the fermentation environment and provided abundant substrates for methane production. This study introduces a promising bio-based strategy to improve the anaerobic digestion efficiency of acidified food waste.

Effect of long-term application of pig slurry and NPK fertilizers on trace metal content in the soil.

One of the goals of sustainable agricultural production is to avoid soil contamination by elements defined as trace metals (TMs). The aim of this study was to assess the long-term impact of the use of pig slurry (PS) and NPK mineral fertilizers on the soil content of cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn). In a 9-year crop rotation, PS was used three times only before root crops. The same four levels of NPK doses (N0P0K0, N1P1K1, N3P2K2, N4P2K2) were applied to both plots with and without PS. Soil samples were collected in early spring from topsoil (0-0.3 m) and subsoil (0.3-0.6 m). Three forms of TMs were determined in the soil: pseudo-total (Aqua regia); bioavailable (Mehlich 3 method) and readily bioavailable (mobile) forms (1 M NH4NO3). The tested factors did not have a significant impact on the Cd, Cu and Pb content, regardless of the form analyzed and the soil depth. PS application significantly increased the content of bioavailable forms of Zn regardless of the year, and the content of pseudo-total Zn only in the sugar beet year, i.e. after manure application. Increasing NPK doses increased the content of mobile Zn in the topsoil, especially in PS plots. A tendency to accumulate mobile forms of Cd and Pb was also observed on NPK-fertilized plots. Thus, long-term application of high NPK doses may increase the risk of contamination of the food chain with these metals. The content of mobile Cd and Zn was positively related to the content of total nitrogen in the soil and negatively related to pH.

Exploring the leaf regeneration cycles response of Zostera japonica to ocean acidification.

Ocean acidification is one of the major global environmental problems facing humankind today, and it has far-reaching impacts on marine organisms and the entire marine ecosystem. Zostera japonica, an important supporting species of intertidal seagrass beds, exhibits high photosynthetic productivity and plays an important role in the carbon cycle of nearshore waters. However, little is known about the characteristics, processes, and mechanisms of its response to ocean acidification. In this study, we conducted a 120-day acidification experiment in Z. japonica; here, plants underwent four leaf regeneration cycles to reveal the response mechanism of Z. japonica to ocean acidification (OA). We found that acidification significantly affected the seedling stage of Z. japonica, impacting leaf regeneration cycles by altering physiological and molecular responses. In one leaf regeneration cycle, the short-term exposure to CO2 affected the seagrass parameters, such as the regulation of inorganic carbon uptake modes and the regulation of photosynthesis between the dark and light reactions, with the potential to affect the carbon sinks of the marine organisms. The long-term effects on the regulation of antioxidant enzymes and antioxidant metabolites, caused an improvement in the marine life adaptation to OA. In a comparison of the different leaf regeneration cycles, the response pattern of Z. japonica showed an offset of the acidification during the short cycles and an adaption to the acidification during the long cycles. This study revealed the response mechanism of Z. japonica to OA at different time scales and could provide a theoretical basis for accurately assessing the impact of OA on seagrass and the entire seagrass bed ecosystem.

Ratiometric Fluorescent pH Sensing with Carbon Dots: Fluorescence Mapping across pH Levels for Potential Underwater Applications.

Ocean acidification has become a major climate change concern requiring continuous observation. Additionally, in the industry, pH surveillance is of great importance. Consequently, there is a pressing demand to develop robust and inexpensive pH sensors. Ratiometric fluorescence pH sensing stands out as a promising concept. The application of carbon dots in fluorescent sensing presents a compelling avenue for the advancement of pH-sensing solutions. This potential is underpinned by the affordability of carbon dots, their straightforward manufacturing process, low toxicity, and minimal susceptibility to photobleaching. Thus, investigating novel carbon dots is essential to identify optimal pH-sensitive candidates. In this study, five carbon dots were synthesized through a simple solvothermal treatment, and their fluorescence was examined as a function of pH within the range of 5-9, across an excitation range of 200-550 nm and an emission range of 250-750 nm. The resulting optical features showed that all five carbon dots exhibited pH sensitivity in both the UV and visible regions. One type of carbon dot, synthesized from m-phenylenediamine, displayed ratiometric properties at four excitation wavelengths, with the best results observed when excited in the visible spectrum at 475 nm. Indeed, these carbon dots exhibited good linearity over pH values of 6-9 in aqueous Carmody buffer solution by calculating the ratio of the green emission band at 525 nm to the orange one at 630 nm (I525nm/I630nm), demonstrating highly suitable properties for ratiometric sensing.

Evolution of chitin-synthase in molluscs and their response to ocean acidification.

Chitin-synthase (CHS) is found in most eukaryotes and has a complex evolutionary history. Research into CHS has mainly been in the context of biomineralization of mollusc shells an area of high interest due to the consequences of ocean acidification. Exploration of CHS at the genomic level in molluscs, the evolution of isoforms, their tissue distribution, and response to environmental challenges are largely unknown. Exploiting the extensive molecular resources for mollusc species it is revealed that bivalves possess the largest number of CHS genes (12-22) reported to date in eukaryotes. The evolutionary tree constructed at the class level of molluscs indicates four CHS Type II isoforms (A-D) probably existed in the most recent common ancestor, and Type II-A (Type II-A-1/Type II-A-2) and Type II-C (Type II-C-1/Type II-C-2) underwent further differentiation. Non-specific loss of CHS isoforms occurred at the class level, and in some Type II (B-D groups) isoforms the myosin head domain, which is associated with shell formation, was not preserved and highly species-specific tissue expression of CHS isoforms occurred. These observations strongly support the idea of CHS functional diversification with shell biomineralization being one of several important functions. Analysis of transcriptome data uncovered the species-specific potential of CHS isoforms in shell formation and a species-specific response to ocean acidification (OA). The impact of OA was not CHS isoform-dependent although in Mytilus, Type I-B and Type II-D gene expression was down-regulated in both M. galloprovincialis and M. coruscus. In summary, during CHS evolution the gene family expanded in bivalves generating a large diversity of isoforms with different structures and with a ubiquitous tissue distribution suggesting that chitin is involved in many biological functions. These findings provide insight into CHS evolution in molluscs and lay the foundation for research into their function and response to environmental changes.

Effect of D,L methionine and ammonium chloride on urine acidification, urinary fractional excretion of calcium, and blood bicarbonate in clinically healthy goats.

BACKGROUND: Urinary acidification with ammonium chloride (AC) for urolith dissolution is a common treatment for goats with urolithiasis. Studies have reported increased fractional excretion of calcium (FECa) following AC administration, which could increase calcium-based urolithiasis. D,L methionine (MET) may result in similar acidification with less calcium excretion. OBJECTIVE: To compare the effects of orally administered MET and AC on urine and blood pH, FECa, and blood HCO3- concentrations in male goats. METHODS: Prospective, randomized, crossover study. 12 healthy, 5-to-6-month-old Boer-cross wethers were administered 200 mg/kg of AC or MET orally for 14 days with a 7-day washout period between trials. Venous blood and urine samples were collected every 2 days. The effects of treatment and treatment day on urine and blood pH, HCO3-, and FECa were assessed using linear mixed models. RESULTS: Ammonium chloride and MET lowered least squares means (LSM) urine pH on day 6 (LSM, 7.49; 95% CI, 6.44 to 8.54), 8 (LSM, 7.78; 95% CI, 6.73 to 8.83), and 10 (LSM, 7.53; 95% CI, 6.49 to 8.58) when compared to day 0 (LSM, 8.23; 95% CI, 7.18 to 9.28). Some goats' urine indicated acidification (pH < 7.0) in the first phase of the trial; however, for the entire trial, a significant treatment effect was not detected on urine pH, blood pH, blood HCO3- or log10 FECa. CLINICAL RELEVANCE: Ammonium chloride and MET acidified urine of some goats. Dietary cation-anion difference should be considered when treating healthy goats to acidify their urine.

Impacts of ocean acidification and hypoxia on cellular immunity, oxygen consumption and antioxidant status in Mediterranean mussel.

There is growing recognition that the hypoxic regions of the ocean are also becoming more acidic due to increasing levels of global carbon dioxide emissions. The impact of water acidification on marine life is largely unknown, as most previous studies have not taken into account the effects of hypoxia, which may affect how organisms respond to low pH levels. In this study, we experimentally examined the consequences of water acidification in combination with normoxic or hypoxic conditions on cellular immune parameters in Mediterranean mussels. We measured total hemocyte counts in hemolymph, the cellular composition of hemolymph, phagocytosis, reactive oxygen species (ROS) production. General response of the organism was evaluated on the basis of the activity of antioxidant enzymes in the hepatopancreas, as well as respiratory rates over an 8-day exposure period. The mussels were exposed to low pH conditions (7.3), either under normoxic conditions (dissolved oxygen concentration of 8 mg/L) or hypoxic conditions (dissolved oxygen concentration of 2 mg/L). The parameters were assessed at days 1, 3, 6, and 8 of the experiment. Experimental acidification under normoxic conditions reduced THC and ROS production by hemocytes during later stages of exposure, but phagocytic activity (PA) only decreased at day 3 and then recovered. Combined acidification and hypoxia suppressed PA in hemocytes at the beginning of exposure, while hemocyte ROS production and THC decreased by the end of the experiment. The hemolymph cellular composition and activity of antioxidant enzymes were unaffected by acidified conditions under different oxygen regimes, but mussel respiratory rate (RR) decreased with a more significant reduction in oxygen consumption under hypoxia. Mussels showed a relatively high tolerance to acidification in combination with various dissolved oxygen levels, although prolonged acidification exposure led to increased detrimental effects on immunity and metabolism.

Effect of ultrasound-attenuation on technological and functional properties of two strains of Lactiplantibacillus plantarum isolated from table olives.

While probiotics have a wide range of beneficial properties, they can also negatively affect the taste or aroma of foods products by resulting in the phenomenon of post-acidification. Ultrasound (US) is a tool to modulate the metabolism of probiotic bacteria, counteracting post-acidification and improving the performance and functional properties of microorganisms without affecting their viability. The purpose of this paper was to evaluate the effect of 10 different combinations of power (20 and 40 %) and duration (2, 4, 6, 8 and 10 min) of US treatment on two functional strains of Lactiplantibacillus plantarum (c16 and c19) isolated from table olives, with the aim of understanding how, some of the main functional and technological traits (viability, acidification, growth profile under different conditions, antibiotic resistance, viability at pH 2.0 and 0.3 % bile salts), were affected. It was found that the effects were strain dependent, and the best results were obtained for strain c19 in the combinations at 20 % for 8 and 10 min and 40 % for 2 min, where an improvement in functional characteristics was found, with some effects on biofilm stability, inhibition of acidification, without adverse results on some technological properties.

Post-acidification of fermented milk and its molecular regulatory mechanism.

The fermented milk products with lactic acid bacteria (LAB) are widely accepted by consumers. During the chilled-chain transportation and storage, LAB in the product keep producing lactic acid, and this will lead to post-acidification, which can affect the flavor, consumer acceptance and even shelf-life of the product. LAB is the determining factor affecting post-acidification. The acid production pathway in LAB and methods inhibiting post-acidification received widespread attention. This review will focus on the post-acidification from the perspective of fermentation starters, including acid production pathway in LAB, main factors and key enzymes affecting post-acidification. Lactobacillus delbrueckii subsp. bulgaricus is a key bacterial species responsible for post acidification in the fermented milk products. The different species and strains presented various differences in process like acid production, acid resistance and post-acidification. Furthermore, multiple factors, such as milk composition, fermentation temperature, and homogenization, also can influence post-acidification. Lactose transport and utilization pathways, as well as its subsequent products metabolic pathway directly influence the post-acidification. F0F1-ATPase, ß-galactosidase, and lactate dehydrogenase are recognized as important enzymes related to post-acidification. The degree of post-acidification is mainly related to the acid production and acid resistance abilities of the fermentation starters, so the key enzymes related to post-acidification are mostly taking part in these two capacities. Recently, some new post-acidification related biomarker genes were found, providing a reference adjusting post-acidification without affecting fermentation rate and bacteria viability. To clarify the post-acidification mechanism at the molecular level will help control post- acidification.

Microwave-assisted organic acids and green hydrogen production during mixed culture fermentation.

BACKGROUND: The integration of anaerobic digestion into bio-based industries can create synergies that help render anaerobic digestion self-sustaining. Two-stage digesters with separate acidification stages allow for the production of green hydrogen and short-chain fatty acids, which are promising industrial products. Heat shocks can be used to foster the production of these products, the practical applicability of this treatment is often not addressed sufficiently, and the presented work therefore aims to close this gap. METHODS: Batch experiments were conducted in 5 L double-walled tank reactors incubated at 37 °C. Short microwave heat shocks of 25 min duration and exposure times of 5-10 min at 80 °C were performed and compared to oven heat shocks. Pairwise experimental group differences for gas production and chemical parameters were determined using ANOVA and post-hoc tests. High-throughput 16S rRNA gene amplicon sequencing was performed to analyse taxonomic profiles. RESULTS: After heat-shocking the entire seed sludge, the highest hydrogen productivity was observed at a substrate load of 50 g/l with 1.09 mol H2/mol hexose. With 1.01 mol H2/mol hexose, microwave-assisted treatment was not significantly different from oven-based treatments. This study emphasised the better repeatability of heat shocks with microwave-assisted experiments, revealing low variation coefficients averaging 29%. The pre-treatment with microwaves results in a high predictability and a stronger microbial community shift to Clostridia compared to the treatment with the oven. The pre-treatment of heat shocks supported the formation of butyric acid up to 10.8 g/l on average, with a peak of 24.01 g/l at a butyric/acetic acid ratio of 2.0. CONCLUSION: The results support the suitability of using heat shock for the entire seed sludge rather than just a small inoculum, making the process more relevant for industrial applications. The performed microwave-based treatment has proven to be a promising alternative to oven-based treatments, which ultimately may facilitate their implementation into industrial systems. This approach becomes economically sustainable with high-temperature heat pumps with a coefficient of performance (COP) of 4.3.

Reprint: Acclimatization and Adaptive Capacity of Marine Species in a Changing Ocean.

To persist in an ocean changing in temperature, pH and other stressors related to climate change, many marine species will likely need to acclimatize or adapt to avoid extinction. If marine populations possess adequate genetic variation in tolerance to climate change stressors, species might be able to adapt to environmental change. Marine climate change research is moving away from single life stage studies where individuals are directly placed into projected scenarios ('future shock' approach), to focus on the adaptive potential of populations in an ocean that will gradually change over coming decades. This review summarizes studies that consider the adaptive potential of marine invertebrates to climate change stressors and the methods that have been applied to this research, including quantitative genetics, laboratory selection studies and trans- and multigenerational experiments. Phenotypic plasticity is likely to contribute to population persistence providing time for genetic adaptation to occur. Transgenerational and epigenetic effects indicate that the environmental and physiological history of the parents can affect offspring performance. There is a need for long-term, multigenerational experiments to determine the influence of phenotypic plasticity, genetic variation and transgenerational effects on species' capacity to persist in a changing ocean. However, multigenerational studies are only practicable for short generation species. Consideration of multiple morphological and physiological traits, including changes in molecular processes (eg, DNA methylation) and long-term studies that facilitate acclimatization will be essential in making informed predictions of how the seascape and marine communities will be altered by climate change.

Natural acidified marine systems: Lessons and predictions.

Natural acidified marine systems (ASs) are environments with relatively low pH levels due to natural causes such as volcanic activity, geochemical reactions, and biological processes. These systems act as natural laboratories for the study of the effects of ocean acidification, allowing for the observation of long-term ecological and evolutionary responses. Understanding these systems is crucial for predicting the effects of anthropogenic ocean acidification (OA) on marine ecosystems. There are 23 ASs in which scientific research has shown significant parallelisms in their results worldwide, such as the disappearance of calcareous organisms and the loss of species with key ecological functions under OA conditions. Future research should emphasize continuous collaboration among teams, as well as public access to oceanographic and biological data along with the monitoring of environmental variables at each AS. To preserve these areas, it is imperative to employ non-destructive methods and protect them as human heritage sites.

Widespread scope for coral adaptation under combined ocean warming and acidification.

Reef-building coral populations are at serious risk of collapse due to the combined effects of ocean warming and acidification. Nonetheless, many corals show potential to adapt to the changing ocean conditions. Here we examine the broad sense heritability (H2) of coral calcification rates across an ecologically and phylogenetically diverse sampling of eight of the primary reef-building corals across the Indo-Pacific. We show that all eight species exhibit relatively high heritability of calcification rates under combined warming and acidification (0.23-0.56). Furthermore, tolerance to each factor is positively correlated and the two factors do not interact in most of the species, contrary to the idea of trade-offs between temperature and pH sensitivity, and all eight species can co-evolve tolerance to elevated temperature and reduced pH. Using these values together with historical data, we estimate potential increases in thermal tolerance of 1.0-1.7°C over the next 50 years, depending on species. None of these species are probably capable of keeping up with a high global change scenario and climate change mitigation is essential if reefs are to persist. Such estimates are critical for our understanding of how corals may respond to global change, accurately parametrizing modelled responses, and predicting rapid evolution.