Bacterial Degradation of Bacteriostatic Polyphenols, Tannins

Tannin degradation by bacteria has not been studied much as tannins are commonly known to be bacteriostatic due to enzyme inhibition, substrate deprivation, and the enzyme activity on the bacterial cell wall. However, about a handful of bacteria have been found to tolerate certain concentrations of tannin. This study focuses on isolating and identifying bacteria from decaying portions of tree bark for tannase production and effective catalysis of ester bond hydrolysis in tannins. Different concentrations of commercial tannic acid were used as the sole carbon source on mineral salt medium (MSM) agar plates, to test the maximum tolerable concentrations (MTCs) by the isolates. Tannin degradation was confirmed by a visual reading method and bacterial tannase activity and the biodegradation percentage were determined. One particular isolate was identified to have 50 g/L MTC of tannin, with a tannase activity of 51.61 U/mL that is optimum after 96 hours of incubation. The 16s rRNA sequencing results showed that the isolate belonged to Bacillus genus and the resulting bacterial strain isolate was found to be a new strain of Bacillus subtilis which was submitted to GenBank under the accession number MH330408.

Unraveling nitrogen, sulfur and carbon metabolic pathways and microbial community transcriptional responses to substrate deprivation and toxicity stresses in a bioreactor mimicking anoxic brackish coastal sediment conditions

Microbial communities are key drivers of carbon, sulfur and nitrogen cycling in coastal ecosystems, where they are subjected to dynamic shifts in substrate availability and exposure to toxic compounds. However, how these shifts affect microbial interactions and function is poorly understood. Unraveling such microbial community responses is key to understand their environmental distribution and resilience under current and future disturbances. Here, we used metagenomics and metatranscriptomics to investigate microbial community structure and transcriptional responses to prolonged ammonium deprivation and sulfide and nitric oxide toxicity stresses in a controlled bioreactor system mimicking coastal sediment conditions. Candidatus Nitrobium versatile, identified in this study as a sulfide-oxidizing denitrifier, became a rare community member upon ammonium removal. The methanotroph Ca. Methanoperedens nitroreducens showed remarkable resilience to both experimental conditions, dominating transcriptional activity of dissimilatory nitrate reduction to ammonium (DNRA). After the ammonium removal experiment, a novel methanotroph species that we have named Ca. Methylomirabilis tolerans outcompeted Ca. Methylomirabilis lanthanidiphila and the anaerobic ammonium oxidizer (anammox) Ca. Kuenenia stuttgartiensis outcompeted Ca. Scalindua rubra. At the end of the sulfide and nitric oxide experiment, a gammaproteobacterium affiliated to the family Thiohalobacteraceae was enriched and dominated transcriptional activity of sulfide:quinone oxidoreductase. Our results indicate that some community members could be more resilient to stresses than others in coastal ecosystems, leading to dynamic microbial community shifts and novel functional states. Methane and sulfide oxidation could be ecosystem functions preserved across the investigated disturbances, while differing nitrogen cycling pathways might be favored in response to stresses.ImportanceCoastal ecosystems are primary zones of biogeochemical cycling, processing inputs of nutrients both generated in situ and derived from land runoff. Microbial communities that inhabit costal sediments perform these biogeochemical reactions, but microbial responses to dynamic, periodic substrate deprivation and exposure to toxic compounds remain elusive. In this study, we sought to address this knowledge gap in a controlled bioreactor system, unraveling microbial metabolic pathways and monitoring microbial responses to stresses that might occur in costal sediments. We identified key microbial players and shifts in their abundance and transcriptional activity. Our results indicated that methanotrophs were particularly resilient to stresses, sulfide oxidizers differed in resiliency but the community maintained sulfide oxidation function across stresses, and that anaerobic ammonium oxidizing (anammox) bacteria were sensitive to substrate deprivation but could restore activity once favorable conditions were reestablished. These insights will help to understand and predict coastal ecosystem responses to future disturbances.

P0535ATG5 OVEREXPRESSION IN PACS SIGNIFICANTLY IMPROVES CELL-MEDIATED AKI PROTECTION

Background and Aims Acute Kidney Injury (AKI) significantly worsens the prognosis of hospitalized patients. Cell-based strategies have been established as reliable option for improving AKI outcomes in experimental AKI. Own studies performed in recent years utilized Pro-angiogenic Cells (PACs). Autophagy (AP) is commonly regarded as process of en-dogenous self-defense. The AP cascade, which may be stimulated by either substrate deprivation or certain exogenous / endogenous stressors, involves the activation of numerous proteins, the so-called Autophagy-related proteins (Atg proteins). Among these, Atg5 has been suggested to play a key role in augmenting AP. The current study evaluated whether selective Atg5 activation in syngeneic murine PACs may result in improved cell-mediated AKI protection. Method Cultured murine PACs were selectively transfected for Atg5. Successful transfection was verified by detecting red fluorescing cells. AKI was induced in male C57/Bl6N mice (8-12 weeks) by bilateral renal ischemia (IRI - 40 minutes). Transfected cells were i.v. injected post-ischemia. Mice were analyzed 48 hours and 6 weeks later. Results IRI induced significant kidney excretory dysfunction as reflected by higher serum cystatin C levels (48 hours and 6 weeks). Cell administration (either native or after transfection) did not improve AKI outcomes at 48 hours. At 6 weeks, injection of native cells resulted in lower serum cystatin C, this effect was even more pronounced if transcfected cells were applied. Conclusion Together, our data show that selective Atg5 overexpression in murine PACs sub-stantially augments cell-mediated AKI protection in the long-term. Thus, a new strategy for improving AKI protective effects of PACs has been identified.

Metabolic and endocrine changes in acute and chronic critical illness

Critical illness, an extreme form of severe physical stress, is characterized by important endocrine and metabolic changes. The development of critical care medicine has made possible survival from conditions that were previously rapidly fatel, and as a result many patients now enter a prolonged phase of chronic or persistent critical illness. Acute endocrine adaptations are directed towards providing energy and substrates for the vital fight or flight response in the context of exogenous substrate deprivation. Distinct endocrine and metabolic alterations characterize the chronic phase of critical illness, which seems to no longer be solely beneficial and may hamper recovery and rehabilitation. Onset of the stressful event causes an acute activation of pulsatile hormonal release from the anterior pituitary, followed by suppression in the chronic phase of illness, ultimately resolving to normality if recovery occurs.