Contrasting patterns of bacterial communities in the rearing water and gut of Penaeus vannamei in response to exogenous glucose addition

Supplementing exogenous carbon sources is a practical approach to improving shrimp health by manipulating the microbial communities of aquaculture systems. However, little is known about the microbiological processes and mechanisms of these systems. Here, the effects of glucose addition on shrimp growth performance and bacterial communities of the rearing water and the shrimp gut were investigated to address this knowledge gap. The results showed that glucose addition significantly improved the growth and survival of shrimp. Although the α-diversity indices of both bacterioplankton communities and gut microbiota were significantly decreased by adding glucose, both bacterial communities exhibited divergent response patterns to glucose addition. Glucose addition induced a dispersive bacterioplankton community but a more stable gut bacterial community. Bacterial taxa belonging to Ruegeria were significantly enriched by glucose in the guts, especially the operational taxonomic unit 2575 (OTU2575), which showed the highest relative importance to the survival rate and individual weight of shrimp, with the values of 43.8 and 40.6%, respectively. In addition, glucose addition increased the complexity of interspecies interactions within gut bacterial communities and the network nodes from Rhodobacteraceae accounted for higher proportions and linked more with the nodes from other taxa in the glucose addition group than that in control. These findings suggest that glucose addition may provide a more stable gut microbiota for shrimp by increasing the abundance of certain bacterial taxa, such as Ruegeria.

Soil biotest results using Azotobacter bacteria: interpretation problems and possible solutions

The aim of the study. The Azotobacter genus is a well-known bioassay for testing the soil environment quality. A large number of these bacteria is considered as evidence of the ecological well-being of a soil. However, a high number of these microorganisms was found in disturbed ecosystems, which means there is a problem of how to interpret the results of the biotest. Therefore the aim of the study was to clarify the causes of this problem and suggest a possible way to solve it. Location and objects of the study. The study was conducted in West Siberia (Russia) in the Priobskoe plateau (54°53'13.5"N, 82°59'36.7E"). Leached сhernozem with different content of organic matter (mortmass) was studied under the following treatments: 1) permanent fallow, 2) wheat cultivation, annual removal of straw from the field + summer fallow, 3) wheat + left straw + summer fallow, 4) wheat +left straw + green manure fallow, 5) grassland. The content of N-NO3, respectively, was equal to 50, 10, 15, 5, 0 mg/kg. Another object was a site at the mining and processing plant "Denisovsky" in South Yakutia (Russia) (56°46'20.23"N, 124°51'06.95"E). Abandoned for a long time (30 years) after coal mining spoils were studied in two variants: without plants and with well-developed vegetation cover. Total SOC content was 1.8 and 5.7%, N – 0.3 and 0.4 %, respectively. Methodology. The direct sowing of single soil aggregates onto the N-free medium containing glucose as C-source was used to cultivate Azotobacter. Glucose (10 mg/g soil) was added to the soil to activate Azotobacter growth. A live culture of bacteria was introduced into the soil at a dose of 1 ml/g to check the viability of Azotobacter in experimental soils. Main results. The status of the microbial community in situ was observed on microbial landscapes obtained by exposing slides in the undisturbed soil for 30 days. The overgrowth of soil lumps in the specified range of options was 0–80–40–0–0% and after glucose addition – 100–80–80–0%. The activation of Azotobacter growth after glucose addition was inversely proportional to the C: N ratio (between the mortmass and the mineral nitrogen). Live Azotobacter culture under grassland developed 2.5 times slower in comparison with the fallow. Similar patterns were found in the study of the soils developed on the coal mining spoils. Activation of Azotobacter growth by glucose (response to stress) was more pronounced in soils with apparently less favorable environment for bacteria. Conclusion. The reason behind misleading interpretation of Azotobacter biotest results was that the original purpose of the test was to assess fertility of arable soils. This role of the indicator bacterium was previously underestimated. It is known that the arable soil belongs to the category of disturbed ones, and the abundance of Azotobacter may indicate instability in the microbial community of the soil. To expand the capabilities of the biotest, the authors propose to supplement the test with a procedure for evaluating the Azotobacter growth response to experimental stress, e.g. C-substrate addition.

Glucose Addition Enhanced the Advanced Treatment of Coking Wastewater

Biological processes have high removal efficiencies and low operational costs, but the secondary effluent of coking wastewater (CWW), even at a low concentration, is difficult for microorganisms to degrade directly. In this study, glucose was used as a carbon source and co-metabolic substrate for microbial acclimation in order to enhance the advanced treatment of coking wastewater (CWW). The removal performance of the pollutants, especially recalcitrant compounds, was studied and the changes in the microbial community structure after activated sludge acclimation were analyzed. The effect of glucose addition on the secondary biochemical effluent of coking wastewater (SBECW) treatment by the acclimated sludge was further studied by a comparison between the performance of two parallel reactors seeded with the acclimated sludge. Our results showed that the concentrations of chemical oxygen demand (COD), total organic carbon (TOC), and UV absorption at 254 nm (UV254) of the wastewater decreased in the acclimation process. Refractory organic matter, such as polycyclic aromatic hydrocarbons and nitrogen-containing heterocyclics, in the SBECW was effectively degraded by the acclimated sludge. High-throughput sequencing revealed that microbes with a strong ability to degrade recalcitrant compounds were enriched after acclimation, such as Thauera (8.91%), Pseudomonas (3.35%), and Blastocatella (10.76%). Seeded with the acclimated sludge, the reactor with the glucose addition showed higher COD removal efficiencies than the control system without glucose addition (p < 0.05). Collectively, glucose addition enhanced the advanced treatment of coking wastewater (CWW).

Fluorescence quenching-based bodipy-boronic acid linked viologen dual system for potential glucose sensing applications

Purpose The purpose of this study is to develop a non-enzymatic based glucose-sensing platform composed of Bodipy-BBV dual system which can be monitored by a photodetector under the blue LED excitation. Design/methodology/approach The sensor has been developed from a dual system including a fluorescent dye, an aldehyde derivative of boron dipyrromethene (Bodipy) and a quencher, orto-boronic acid linked viologen (o-BBV) where their combination resulted in a ratiometric fluorescence quenching in ethanol: PBS (1:1, pH:7.4) solution under UV light excitation. By glucose addition, o-BBV has been released from the Bodipy and binded to cis-diol groups of glucose, thereby fluorescence emission of Bodipy has been regained. Furthermore, a setup consisting of a light emitting diode (LED) and a photodiode (PD) was used to prove electrical detection of glucose without the need for expensive and bulky optical equipment, enabling the development of a miniaturized and low-cost glucose-sensing platform. Findings The fluorescence intensity of the Bodipy derivative in the solution (2 × 10−6 M) was diminished by 93% in the presence of o-BBV solution (5 × 10−3 M). Upon the glucose addition, 81% of the Bodipy fluorescence intensity has been recovered after introduction of 30 mM of glucose, where the ratio of o-BBV/Bodipy was 35:1. A linear response between 10 and 30 mM glucose concentration was obtained, which covers the biologically significant range. A high correlation between the photodiode current and Bodipy fluorescence intensity was achieved. Originality/value Even though Bodipy molecules are known with their superior optical properties and applied to the fluorescence-based detection of glucose, to the best of the authors’ knowledge, no work has been reported on Bodipy-BBV dual system to detect glucose molecules as a non-enzymatic based method. This design enables the dye and the quencher to independently coexist in the solution, allowing for tuning of their individual concentrations to optimize the glucose sensitivity. Furthermore, an electrical light detection scheme consisting of a LED and a photodiode has been implemented to eliminate the bulky optical equipment from the measurement setup and further this work for the development of a compact and inexpensive sensor. The results presented here demonstrate the feasibility of this system for the development of a novel glucose sensor.

Co-metabolic Effect of Glucose on Methane Production and Phenanthrene Removal in an Enriched Phenanthrene-Degrading Consortium Under Methanogenesis

Anaerobic digestion is used to treat diverse waste classes, and polycyclic aromatic hydrocarbons (PAHs) are a class of refractory compounds that common in wastes treated using anaerobic digestion. In this study, a microbial consortium with the ability to degrade phenanthrene under methanogenesis was enriched from paddy soil to investigate the cometabolic effect of glucose on methane (CH4) production and phenanthrene (a representative PAH) degradation under methanogenic conditions. The addition of glucose enhanced the CH4 production rate (from 0.37 to 2.25mg⋅L−1⋅d−1) but had no influence on the degradation rate of phenanthrene. Moreover, glucose addition significantly decreased the microbial α-diversity (from 2.59 to 1.30) of the enriched consortium but showed no significant effect on the microbial community (R2=0.39, p=0.10), archaeal community (R2=0.48, p=0.10), or functional profile (R2=0.48, p=0.10). The relative abundance of genes involved in the degradation of aromatic compounds showed a decreasing tendency with the addition of glucose, whereas that of genes related to CH4 synthesis was not affected. Additionally, the abundance of genes related to the acetate pathway was the highest among the four types of CH4 synthesis pathways detected in the enriched consortium, which averagely accounted for 48.24% of the total CH4 synthesis pathway, indicating that the acetate pathway is dominant in this phenanthrene-degrading system during methanogenesis. Our results reveal that achieving an ideal effect is diffcult via co-metabolism in a single-stage digestion system of PAH under methanogenesis; thus, other anaerobic systems with higher PAH removal efficiency should be combined with methanogenic digestion, assembling a multistage pattern to enhance the PAH removal rate and CH4 production in anaerobic digestion.

Pak1 kinase controls cell shape through ribonucleoprotein granules

Fission yeast cells maintain a rod shape due to conserved signaling pathways that organize the cytoskeleton for polarized growth. We discovered a mechanism linking the conserved protein kinase Pak1 with cell shape through the RNA-binding protein Sts5. Pak1 (also called Shk1 and Orb2) prevents Sts5 association with P bodies by directly phosphorylating its intrinsically disordered region (IDR). Pak1 and the cell polarity kinase Orb6 both phosphorylate the Sts5 IDR but at distinct residues. Mutations preventing phosphorylation in the Sts5 IDR cause increased P body formation and defects in cell shape and polarity. Unexpectedly, when cells encounter glucose starvation, PKA signaling triggers Pak1 recruitment to stress granules with Sts5. Through retargeting experiments, we reveal that Pak1 localizes to stress granules to promote rapid dissolution of Sts5 upon glucose addition. Our work reveals a new role for Pak1 in regulating cell shape through ribonucleoprotein granules during normal and stressed growth conditions.

Multi-omics analysis of glucose-mediated signaling by a moonlighting Gβ protein Asc1/RACK1

Heterotrimeric G proteins were originally discovered through efforts to understand the effects of hormones, such as glucagon and epinephrine, on glucose metabolism. On the other hand, many cellular metabolites, including glucose, serve as ligands for G protein-coupled receptors. Here we investigate the consequences of glucose-mediated receptor signaling, and in particular the role of a Gα subunit Gpa2 and a non-canonical Gβ subunit, known as Asc1 in yeast and RACK1 in animals. Asc1/RACK1 is of particular interest because it has multiple, seemingly unrelated, functions in the cell. The existence of such “moonlighting” operations has complicated the determination of phenotype from genotype. Through a comparative analysis of individual gene deletion mutants, and by integrating transcriptomics and metabolomics measurements, we have determined the relative contributions of the Gα and Gβ protein subunits to glucose-initiated processes in yeast. We determined that Gpa2 is primarily involved in regulating carbohydrate metabolism while Asc1 is primarily involved in amino acid metabolism. Both proteins are involved in regulating purine metabolism. Of the two subunits, Gpa2 regulates a greater number of gene transcripts and was particularly important in determining the amplitude of response to glucose addition. We conclude that the two G protein subunits regulate distinct but complementary processes downstream of the glucose-sensing receptor, as well as processes that lead ultimately to changes in cell growth and metabolism.

The effect of inoculum and glucose addition on polyhydroxyalkanoate production by Brevibacterium sp. B45

Petroleum-based plastics are the major cause of environmental pollution because the plastics need years to be degraded. The difficulties in handling waste of petroleum-based plastic have motivated researchers to produce environmentally friendly plastic materials that are biologically degradable; one of them is polyhydroxyalkanoate (PHA). Polyhydroxyalkanoate is natural biodegradable biopolymers produced by bacteria as an intracellular carbon and energy storage. This polymer is an alternative source of plastics with similar physical properties to petroleum-based plastic.It can be easily biodegraded aerobically and anaerobically. This study examined the potential of one superior isolate as PHA producers, i.e.,Brevibacterium sp. B45. Brevibacterium sp. B45 was cultivated in Ramsay’s minimal medium with inoculum concentrations were 1, 2, and 3% (v/v)and glucose concentrations were 1, 3, and 5% (w/v). The cultivation of Brevibacterium sp. B45 was carried out in a 500 mL Erlenmeyer flask on a shaker incubator with 150 rpm and 30 oC for 72 hours. PHA recovery was carried out by chloroform extraction and characterized by scanning electron microscopy (SEM), Fourier transformed infrared (FTIR), and differential scanning calorimetric (DSC) methods. The highest yield of dried biomass (2.92%) was obtained using 3% inoculum and 3% glucose. The melting temperature (Tm), enthalpy (ΔHf), and crystallinity (Xc) of the PHA product were 172.1 °C, 61.04 J g-1, and 41.08%, respectively. Data of SEM show that a porous surface characterized morphological of purified PHA grains. The functional units of purified PHA grains were C=O, CH3, C-O, C-O-C, C-C, C-H, and -OH. The purified PHA grains show a similar spectrum to the standard Poly-3-hydroxybutyrate (PHB). Therefore, it could be assumed that PHA produced by Brevibacterium sp. B45 was most likely PHB.