Effect of Remediation Reagents on Bacterial Composition and Ecological Function in Black-Odorous Water Sediments

Black-odorous urban water bodies and sediments pose a serious environmental problem. Herein, we conducted microcosm batch experiments to investigate the effect of remediation reagents (magnesium hydroxide and calcium nitrate) on native bacterial communities and their ecological functions in the black-odorous sediment of urban water. The dominant phyla (Proteobacteria, Actinobacteria, Chloroflexi, and Planctomycetes) and classes (Alpha-, Beta-, and Gamma-proteobacteria, Actinobacteria, Anaerolineae, and Planctomycetia) were determined under calcium nitrate and magnesium hydroxide treatments. Functional groups related to aerobic metabolism, including aerobic chemoheterotrophy, dark sulfide oxidation, and correlated dominant genera (Thiobacillus, Lysobacter, Gp16, and Gaiella) became more abundant under calcium nitrate treatment, whereas functional genes potentially involved in dissimilatory sulfate reduction became less abundant. The relative abundance of chloroplasts, fermentation, and correlated genera (Desulfomonile and unclassified Cyanobacteria) decreased under magnesium hydroxide treatment. These results indicated that calcium nitrate addition improved hypoxia-related reducing conditions in the sediment and promoted aerobic chemoheterotrophy.

Transcriptome provides potential insights into how calcium affects the formation of stone cell in Pyrus

Background The content of stone cells in pears has a great influence on taste. Stone cells are formed by the accumulation of lignin. The treatment of exogenous calcium can affect the lignin synthesis, but this Ca-mediated mechanism is still unclear. In this study, the author performed a comparative transcriptomic analysis of callus of pears (Pyrus x bretschneideri) treated with calcium nitrate Ca (NO3)2 to investigate the role of calcium in lignin synthesis. Results There were 2889 differentially expressed genes (DEGs) detected between the Control and Ca (NO3)2 treatment in total. Among these 2889 DEGs, not only a large number of genes related to Ca single were found, but also many genes were enriched in secondary metabolic pathway, especially in lignin synthesis. Most of them were up-regulated during the development of callus after Ca (NO3)2 treatment. In order to further explore how calcium nitrate treatment affects lignin synthesis, the author screened genes associated with transduction of calcium signal in DEGs, and finally found CAM, CML, CDPK, CBL and CIPK. Then the author identified the PbCML3 in pears and conducted relevant experiments finding the overexpression of PbCML3 would increase the content of pear stone cells, providing potential insights into how Ca treatment enhances the stone cell in pears. Conclusions Our deep analysis reveals the effects of exogenous calcium on calcium signal and lignin biosynthesis pathway. The function of PbCML3 on stone cells formation was verified in pear.

The Influence of Acid Whey on the Lipid Composition and Oxidative Stability of Organic Uncured Fermented Bacon after Production and During Chilling Storage

The aim of this research was to evaluate the effect of acid whey on changes in the fatty acid profile, oxidative stability, physico-chemical parameters, and microbiological and sensory quality of traditional organic uncured fermented Polish bacon after production and during chilling storage. Three different treatments of fermented bacon were produced: C—control bacon with a nitrite curing mixture; T—bacon with a nitrate curing mixture; and AW—bacon with acid whey and NaCl. The acid whey used in the production of uncured fermented pork bacon positively changed the sensorial characteristics, directly after the ripening process, and had a positive effect in terms of a decrease in the pH of the product. All of the fermented bacon treatments in general were of good microbiological quality. A higher lactic acid bacteria (LAB) level was observed in the AW treatment after the fermentation process, and the bacteria number did not change during storage, whereas in the C and T treatments, the LAB level increased during storage (p < 0.05). The application of acid whey did not limit the formation of secondary oxidation products (TBARS) during bacon ripening (1.68 mg MDA kg−1), but had a reduced value during storage time (0.73 mg MDA kg−1). The highest polyunsaturated fatty acid (PUFA) levels, after ripening and after four weeks of refrigerated storage, were found in the C treatment. In the AW treatment, it was found that the PUFA level increased; likewise, the content of n-3 and n-6 fatty acids increased, while saturated fatty acids (SFAs) decreased during storage (p < 0.05). The opposite tendency was observed in the C treatment. After four weeks of storage, the PUFA/SFA ratio was the lowest in the nitrate treatment, and higher values of the PUFA/SFA ratio were obtained in the acid whey and nitrite treatment (p < 0.05).

Detection of sulfate-reducing bacteria as an indicator for successful mitigation of sulfide production

Sulfate-reducing bacteria (SRB) are one of the main sources of biogenic H 2 S generation in oil reservoirs. Excess H 2 S production in these systems leads to oil biosouring, which causes operational risks, health hazards and can increase the cost of refining crude oil. Nitrate salts are often added to the system to suppress sulfidogenesis. Because SRB populations can persist in biofilms even after nitrate treatment, identifying shifts in the sessile community is crucial for successful mitigation. However, sampling the sessile community is hampered by its inaccessibility. Here we use the results of a long-term (148 days) ex situ experiment to identify particular sessile community members from observations of the sample waste stream. Microbial community structure was determined for 731 samples across twenty bioreactors using 16S rRNA gene sequencing. By associating microbial community structure with specific steps in the mitigation process, we could distinguish between taxa associated with H 2 S production and mitigation. After initiation of nitrate treatment, certain SRB populations increased in the planktonic community during critical time points, indicating the dissociation of SRBs from the biofilm. Predicted relative abundances of the dissimilatory sulfate reduction pathway also increased during the critical time points. Here, by analyzing the planktonic community structure, we describe a general method that uses high-throughput amplicon sequencing, metabolic inferences, and cell abundance data to identify successful biofilm mitigation. We anticipate that our approach is also applicable to other systems where biofilms must be mitigated but cannot be easily sampled. Importance Microbial biofilms are commonly present in many industrial processes and can negatively impact performance and safety. Within the oil industry, subterranean biofilms cause biosouring with implications for oil quality, cost, occupational health, and the environment. Because these biofilms cannot be directly sampled, methods are needed to indirectly assess the success of mitigation measures. This study demonstrates how the planktonic microbial community can be used to assess the dissociation of SRB-containing biofilms. We found that an increase in the abundance of a specific SRB population in the effluent after nitrate treatment can be used as a potential indicator for the successful mitigation of biofilm-forming SRBs. Moreover, a method for determining critical time points for detecting potential indicators is suggested. This study expands our knowledge in improving mitigation strategies for biosouring and could have broader implications in other systems where biofilms lead to adverse consequences.

Non-Deep Simple Morphophysiological Dormancy and Germination Characteristics of Gentiana triflora var. japonica (Kusn.) H. Hara (Gentianaceae), a Rare Perennial Herb in Korea

This study investigated the kind of seed dormancy and seed germination of Gentiana triflora var. japonica (Kusn.) H. Hara for developing a seed propagation method. The seeds were collected in October 2020 from plants at Mt. Sobaeksan, Korea. In a water imbibition experiment, seed weights increased by > 101.9% of their initial masses over 12 h. Effects of incubation temperature (5, 15, 20, 25, 15/6, or 25/15 °C), cold stratification period (5 °C; 0, 4, 8, or 12 weeks), and gibberellic acid (GA3; 0, 10, 100, or 1000 mg∙L−1) and potassium nitrate treatment (KNO3; 0, 1000, 2000, or 4000 mg∙L−1) on seed germination were investigated to characterize seed dormancy. These seeds exhibited underdeveloped embryos during seed dispersal. The seeds failed to reach the final germination of 15.0% after treatment at 5, 15, 20, 25, 15/6, or 25/15 °C. After cold stratification for 8 weeks, the germination increased dramatically by > 90.0% compared to that at 0 weeks. After the GA3 treatment, the germination reached > 80.0% within 5 days. The final germination was 90.0% in the 100 mg∙L−1 GA3 treatment group. However, the KNO3 treatment had no effect on seed germination. Therefore, the G. triflora var. japonica seeds exhibited non-deep simple morphophysiological dormancy.

In vivo imaging and quantification of carbon tracer dynamics in nodulated root systems of pea plants

Legumes associate with root colonizing rhizobia that provide fixed nitrogen to its plant host in exchange for recently fixed carbon. There is a lack in understanding how individual plants modulate carbon allocation to a nodulated root system as a dynamic response to abiotic stimuli. One reason is that most approaches are based on destructive sampling, making quantification of localized carbon allocation dynamics in the root system difficult. We established an experimental workflow for routinely using non-invasive Positron Emission Tomography (PET) to follow the allocation of leaf-supplied 11C tracer towards individual nodules in a three-dimensional (3D) root system of pea (Pisum sativum). Nitrate was used for triggering the shutdown of biological nitrogen fixation (BNF) expected to rapidly affect carbon allocation dynamics in the root-nodule system. This nitrate treatment lead to a reduction of 11C tracer allocation to nodules by 40% - 47% in 5 treated plants while the variation in control plants was less than 11%. The established experimental pipeline enabled for the first time that several plants could consistently be labelled and measured using 11C tracer in a PET approach to quantify C-allocation to individual nodules following a BNF shutdown. This demonstrates the strength of using 11C tracers in a PET approach for non-invasive quantification of dynamic carbon allocation in several growing plants over several days. A major advantage of the approach is the possibility to investigate carbon dynamics in small regions of interest in a 3D system such as nodules in comparison to whole plant development.

New Insights of Potassium Sources Impacts as Foliar Application on ‘Canino’ Apricot Fruit Yield, Fruit Anatomy, Quality and Storability

This is the first report to study the impacts of potassium sources on apricot fruit yield, quality and storability as a preharvest foliar application. Five sources of potassium (K-humate, K-sulphate, K-nitrate, K-silicate and K-citrate), plus water as a control treatment, were applied individually at 0.2% three times on ‘Canino’ apricot over the 2019 and 2020 seasons. The results showed that all potassium salts, applied foliarly, have potential to improve yield, fruit color, and some fruit physical attributes, such as: weight, size and firmness, as well as a reduced lipid peroxidation, accompanied by a low fruit malondialdehyde content reflected in a high tolerance during storage. The K-nitrate treatment was more effective in the improvement of fruit yield, preharvest quality parameters and keeping fruit postharvest quality characteristics from sharp decline during cold storage. Concerning fruit anatomy, K-nitrate and K-citrate showed thicker cuticle and epidermal parenchyma cell diameters, while the K-silicate induced the highest cell wall thickness. K-nitrate was the most economical, and could be recommended for apricot growers in the Nubaria region of Egypt.

Dietary Nitrate Supplementation Alters Protein and Lipid Metabolism in Zebrafish (Danio rerio) Livers

Objectives Dietary nitrate supplementation shows protective effects against cardio-metabolic disease, decreases pulmonary oxygen uptake, and improves exercise performance in animal models and humans. However, the biological effect of nitrate on energy metabolism in the liver is not well understood. The objective of this study was to elucidate changes in liver metabolism associated with nitrate treatment and exercise. Methods Fish were exposed to sodium nitrate (606.9 mg/L), or control water, for 21 days and analyzed at intervals during a strenuous exercise test. We utilized untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis to determine the effect of nitrate treatment and exercise on the liver metabolome. We measured gene expression of 31 genes linked to energy metabolism and redox signaling. Results In the absence of exercise, nitrate treatment upregulated expression of genes central to nutrient sensing (pgc1a and sirt3), protein synthesis (mtor) and purine metabolism (pnp5a and ampd1) and downregulated expression of genes involved in mitochondrial fat oxidation (acaca, cpt2 and hadh). Upregulation of these genes was associated with an increased abundance of metabolites involved in endogenous nitric oxide metabolism, dopamine biosynthesis, branched chain amino acid metabolism, and lipid metabolism in nitrate-treated livers at rest, compared to rested controls. As expected, the availability of these metabolites was diminished in nitrate-treated livers relative to rested controls. We found no significant change in gene of metabolites directly linked to glycolysis. Conclusions The main novel finding of this study was that sub-chronic nitrate treatment altered dopamine biosynthesis, protein synthesis and lipid metabolism in zebrafish liver without exercise. This is significant because dietary nitrate is emerging as an interesting therapeutic modality for metabolic syndrome and non-alcoholic fatty liver disease by preventing lipid accumulation in the liver. Funding Sources Celia Strickland and G. Kenneth Austin III Endowment and National Institutes of Health.

Nitrate-Induced Improvements in Exercise Performance Is Coincident With Exuberant Changes in Metabolic Genes and the Metabolome in Zebrafish (Danio rerio) Skeletal Muscle

Objectives Dietary nitrate (NO3−) supplementation improves exercise performance by reducing the oxygen cost of exercise and enhancing skeletal muscle function. However, the mechanisms underlying the beneficial effects on exercise performance are not well understood and may be supported by changes in metabolism within the skeletal muscle. The purpose of this study was to elucidate nitrate-induced changes in skeletal muscle energy metabolism associated with improvements in exercise performance that may reflect enhanced metabolic flexibility. Methods Fish were exposed to sodium nitrate (60.7 mg/L, 303.5 mg/L, and 606.9 mg/L), or control water, for 21 days and analyzed at intervals during a strenuous exercise test. Nitrate storage in muscle was measured using chemiluminescence. We utilized nuclear magnetic resonance spectroscopy (NMR), liquid-chromatography tandem mass spectrometry (LC-MS/MS) untargeted metabolomics and real-time quantitative polymerase chain reaction (RT-qPCR) to determine changes in muscle metabolism with nitrate and exercise. Results Nitrate treatment significantly increased muscle nitrate concentrations, while muscle nitrate levels declined with increasing exercise duration, and nitrate treatment was associated with a decrease in the oxygen cost of exercise. In skeletal muscle, nitrate treatment upregulated expression of genes central to nutrient sensing (mtor), glucose (hk2) and lipid metabolism (acaca), redox signaling (nrf2a) and muscle differentiation (sox6). Nitrate treatment caused rested skeletal muscle to have significantly increased metabolites directly linked to energy production (phosphocreatine (PCr), creatine (Cr), adenosine nucleosides, purines, glycolytic, fatty acid and tricarboxylic acid cycle (TCA) intermediates) and a concomitant decrease in these metabolites after exercise, compared to rested-control fish. Conclusions Our data suggest that nitrate exposure may improve exercise performance by changing the metabolic programming of muscle prior to exercise, thus increasing the availability of energy producing metabolites required for exercise such as ATP and phosphocreatine. Funding Sources Celia Strickland and G. Kenneth Austin III Endowment and National Institutes of Health.