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Applying Dialysis Bags to Grow Microalgae and Measure Grazing Rates by Secondary Producers.

ABSTRACT: Traditional methods using sealed bottles to determine the grazing rates by secondary producers neglect chemical changes induced by biological activities during the incubation, giving rise to instable levels of nutrients, pH, pCO₂, pO₂ and other chemicals along with changing microalgal cell concentrations and grazers' metabolism. Here, we used dialysis bags, which allows exchanges of nutrients and gases, to grow microalgae and to determine grazing rates of secondary producers. The specific growth rate of diatom within the dialysis bags increased with increasing water velocities, indicating its suitability to grow microalgae under dynamic water conditions. Then, we compared the grazing rates by the heterotrophic dinoflagellate Noctiluca scintillans measured with the traditional method using polycarbonate (PC) bottles and the approach with the dialysis bags, and found that these two methods gave rise to comparable grazing rates. Nevertheless, the concentrations of inorganic nitrogen and phosphate in the closed PC bottles were about 89-94% lower than those in the dialysis bags due to the microalga's assimilation. Subsequently, we applied it to determine the grazing rates by a copepod and an oyster (in the presence of other grazers). Consistent results were obtained using the dialysis bags to determine grazing rates by copepods. During the mesocosm (3000 L) experiment in the presence of primary and secondary producers, the grazing rates by the oyster Crassostrea angulata were determined based on the difference of phytoplankton biomass within and outside of the dialysis bags that held all organisms in the mesocosm except the oyster. Since the dialysis bags are permeable to gases, the grazing rates by the oyster under 410 (AC) and 1,000 (HC) μatm CO₂ were successfully measured, with a promising result that HC significantly increased the oyster's grazing. We concluded that using dialysis bags to grow microalgae and to determine grazing rates is a reliable approach, especially under different levels of CO₂ and O₂.

SUBMITTER: Tian Y

PROVIDER: S-EPMC9127382 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Background and objectivesRapid ultrafiltration rates are associated with adverse outcomes among patients on hemodialysis. The Centers for Medicare and Medicaid Services is considering an ultrafiltration rate quality measure for the ESRD Quality Incentive Program. Two measure developers proposed ultrafiltration rate measures with different selection criteria and specifications. We aimed to compare the proposed ultrafiltration rate measures and quantify dialysis facility operational burden if treatment times were extended to lower ultrafiltration rates.Design, setting, participants, & measurementsData were taken from the 2012 database of a large dialysis organization. Analyses of the Centers for Medicare and Medicaid Services measure considered 148,950 patients on hemodialysis, and analyses of the Kidney Care Quality Alliance measure considered 151,937 patients. We described monthly patient and facility ultrafiltration rates and examined differences in patient characteristics across ultrafiltration rate thresholds and differences in facilities across ultrafiltration rate measure scores. We computed the additional treatment time required to lower ultrafiltration rates <13 ml/h per kilogram.ResultsUltrafiltration rates peaked in winter and nadired in summer. Patients with higher ultrafiltration rates were younger; more likely to be women, nonblack, Hispanic, and lighter in weight; and more likely to have histories of heart failure compared with patients with lower ultrafiltration rates. Facilities had, on average, 20.8%±10.3% (July) to 22.8%±10.6% (February) of patients with ultrafiltration rates >13 ml/h per kilogram by the Centers for Medicare and Medicaid Services monthly measure. Facilities had, on average, 15.8%±8.2% of patients with ultrafiltration rates ≥13 ml/h per kilogram by the Kidney Care Quality Alliance annual measure. Larger facilities (>100 patients) would require, on average, 33 additional treatment hours per week to lower all facility ultrafiltration rates <13 ml/h per kilogram when total treatment time is capped at 4 hours.ConclusionsUltrafiltration rates vary seasonally and across clinical subgroups. Extension of treatment time as a strategy to lower ultrafiltration rates may pose facility operational challenges. Prospective studies of ultrafiltration rate threshold implementation are needed.

Project description:Microbial secondary metabolites represent a rich source of valuable compounds with a variety of applications in medicine or agriculture. Effective exploitation of this wealth of chemicals requires the functional expression of the respective biosynthetic genes in amenable heterologous hosts. We have previously established the TREX system which facilitates the transfer, integration and expression of biosynthetic gene clusters in various bacterial hosts. Here, we describe the yTREX system, a new tool adapted for one-step yeast recombinational cloning of gene clusters. We show that with yTREX, Pseudomonas putida secondary metabolite production strains can rapidly be constructed by random targeting of chromosomal promoters by Tn5 transposition. Feasibility of this approach was corroborated by prodigiosin production after yTREX cloning, transfer and expression of the respective biosynthesis genes from Serratia marcescens. Furthermore, the applicability of the system for effective pathway rerouting by gene cluster adaptation was demonstrated using the violacein biosynthesis gene cluster from Chromobacterium violaceum, producing pathway metabolites violacein, deoxyviolacein, prodeoxyviolacein, and deoxychromoviridans. Clones producing both prodigiosin and violaceins could be readily identified among clones obtained after random chromosomal integration by their strong color-phenotype. Finally, the addition of a promoter-less reporter gene enabled facile detection also of phenazine-producing clones after transfer of the respective phenazine-1-carboxylic acid biosynthesis genes from Pseudomonas aeruginosa. All compounds accumulated to substantial titers in the mg range. We thus corroborate here the suitability of P. putida for the biosynthesis of diverse natural products, and demonstrate that the yTREX system effectively enables the rapid generation of secondary metabolite producing bacteria by activation of heterologous gene clusters, applicable for natural compound discovery and combinatorial biosynthesis.

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Applying Dialysis Bags to Grow Microalgae and Measure Grazing Rates by Secondary Producers.

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