Respirometric Assessment Research Articles

Ecotoxicological assessment of natural soil amended with sewage sludge: the impacts on soil edaphic organisms and microbial community.

Sewage sludge usage as agricultural soil amendment is a well-known practice employed worldwide. However, certain components may pose risks to the soil ecosystem. For a better verification of the potential adverse effects on the soil biota, biological assays have become an indispensable tool for an accurate understanding of the residue's behavior on soil, as well as its potential toxicity. Accordingly, to properly assess the effects of natural tropical soil (Oxisoil) amended with sewage sludge, we conducted toxicological tests with edaphic organisms (Enchytraeus crypticus and Folsomia candida) and microbial biomass (through respirometric assessment). Results indicate that E. crypticus and F. candida present similar reproduction sensitivity behavior to sewage sludge. For the microbiological analysis, the results suggest that microbial activity was stimulated by sludge application. For further evaluation of respiration of the microbial community and CO2 stabilization values behavior, Ford-Walford modeling was applied and presented limit values for sludge application in soil for 1.5gkg-1 and 15.0gkg-1 of, approximately, 55mg and 88mg, respectively. CO2 releases were faster and reached stability within 18weeks for the soil with higher sludge content (15.0gkg-1 of dry soil). In contrast, CO2 releases were slower for the soil with lower sludge content (1.5gkg-1 of dry soil), and until the experiment's final period (21weeks) respiration behavior did not reach stability. This study indicates that the stabilized sewage sludge, at the considered recommended application rate, presents a low toxicity risk for the studied bioindicators, being suitable for agricultural use.

Respirometric assessment of bacterial kinetics in algae-bacteria and activated sludge processes

Algae-bacteria (AB) consortia can be exploited for effective wastewater treatment, based on photosynthetic oxygenation to reduce energy requirements for aeration. While algal kinetics have been extensively evaluated, bacterial kinetics in AB systems are still based on parameters taken from the activated sludge models, lacking an experimental validation for AB consortia. A respirometric procedure was therefore proposed, to estimate bacterial kinetics in both activated sludge and AB, under different conditions of temperature, pH, dissolved oxygen, and substrate availability. Bacterial activities were differently influenced by operational/environmental conditions, suggesting that the adoption of typical activated sludge parameters could be inadequate for AB modelling. Indeed, respirometric results show that bacteria in AB consortia were adapted to a wider range of conditions, compared to activated sludge, confirming that a dedicated calibration of bacterial kinetics is essential for effectively modelling AB systems, and respirometry was proven to be a powerful and reliable tool to this purpose.

Effect of Temperature Variations on the Oxygen Consumption of the Aerobic Biomass in a Uasb-Mbr System -Respirometric Assessment

Effect of Temperature Variations on the Oxygen Consumption of the Aerobic Biomass in a Uasb-Mbr System -Respirometric Assessment

Particle size distribution of chemical oxygen demand in industrial effluents: impact on effective filtration size and modelling of membrane bioreactors

AbstractThis study reviews particle size distribution (PSD) analysis for industrial effluents. Evaluation of available experimental data indicated that the approach started with arbitrary selection of size distribution. Following a pioneering study on pulp and paper effluent, sequential filtration and ultrafiltration has established itself as a prescribed methodology complementing respirometric assessment of chemical oxygen demand (COD) fractionation based on biodegradation characteristics. Results indicated that most industrial effluents exhibited a COD distribution where soluble fractions accounted for the majority of the total COD, with significant implications for the understanding of membrane bioreactor (MBR) systems. Observations on the accumulation of soluble COD in the reactor to levels much higher than the permeate COD paved the way to define an experimental PSD methodology for assessing the effective filtration size for the membrane module. This novel parameter considered in the light of reported PSD analyses showed that a portion of what was conventionally considered soluble COD would be retained by the size barrier of the membrane and recycled back to the reactor. Thus, MBR systems require a new mathematical model structure that would include a modified COD fractionation, account for the retained COD fractions as additional model components and utilize related mass balance relationships in order to yield an accurate mechanistic description of microbial reactions taking place in the system. © 2021 Society of Chemical Industry (SCI).

Effects of biostimulation by sugarcane bagasse and coffee grounds on sewage sludges, focusing agricultural use: Microbial characterization, respirometric assessment and toxicity reduction

Sewage sludge (SS) exhibits a relevant agronomic potential due to the high content of organic matter and nutrients. However, the presence of several toxic substances can prevent its agricultural application. This study evaluated if the incorporation of stimulating agents (coffee grounds and sugarcane bagasse) could contribute to an effective increase of the SS biodegradability in order to decrease its toxicity. The samples were prepared mixing aerobic or anaerobic sludge with soil, soil and bagasse, and soil and coffee grounds. Respirometric tests showed that stimulating agents enhanced the CO2 production. However, in terms of biodegradation efficiency, more satisfactory results were verified for the anaerobic SS, especially when mixed with coffee grounds. The biodegradation also favored the SS sanitization, eliminating the Enterobacteria. For baseline toxicity (Microtox with Aliivibrio fischeri) and phytotoxicity (Lactuca sativa), all the initial samples showed higher effects. Nevertheless, after the biodegradation, this toxicity was significantly decreased and the best results were obtained for the mixtures containing only soil and sludge. For the AREc32 assay (NRF2 mediated oxidative stress response), although a very weak response was observed, this effect was attenuated for the aerobic SS or completely eliminated for the anaerobic SS after the biodegradation. Thus, even though the use of biostimulation agents during the biodegradation led to an enhancement of microbial respiration, their incorporation to the samples do not seem to interfere in the decrease of the toxic potential of the studied SSs. However, the SS biodegradation in aerobiosis was crucial for toxicity reduction and to accelerate its maturity.

Respirometric assessment of vegetable tanning process wastewater generated from tanneries

AbstractRegistration, Evaluation, and Authorization of CHemicals (REACH) regulations insist on ready biodegradability information for organic chemicals that are used on a large scale in industries. Leather industry being one such, involves addition of organic chemicals that are obtained from plant extracts for vegetable tanning process. In view of the fact that biodegradability assessment is of paramount importance now for industrial wastewaters, a study was conducted on vegetable tanning process wastewater generated from tanneries. As per the Organization for Economic Cooperation and Development (OECD) methods for ready biodegradability (301 C), respirometer studies were conducted on wastewater containing different tannin concentrations of 540 mg/L (T1), 900 mg/L (T2), and 1350 mg/L (T3). The oxygen uptake rate had shown a sharp initial peak at 74, 102, and 164 mg O2/d for the three samples T1, T2, and T3, respectively. The primary degradation % of tannins was found to be 78%, 50%, and 14% for T1, T2, and T3 respectively. The Chemical Oxygen Demand and Biochemical Oxygen Demand removal efficiency also decreased with increase in tannin concentration. The t1/2 values of tannin, calculated using the first‐order kinetic equation, was 8.8, 10.9, and 12.4 days and the experimentally observed t1/2 values were 9.8, 11.5, and 12.6 days for T1, T2, and T3 respectively. This study establishes that presence of tannins in the aerobic treatment significantly affect the oxygen uptake when the tannin concentration is increased. This study emphasizes that ready biodegradability assessment is essential to establish biodegradability of all the organic chemicals used during leather making process.

Comparison of two anti-diabetic monoestolides regarding effects on intact murine liver tissue

Context Monoestolides belonging to the fatty acid-hydroxy fatty acid (FAHFA) family have recently emerged as promising insulin sensitizers. Objective To investigate and compare impact of two selected FAHFA isomers, namely 9-hexadecanoyloxy-octadecanoic acid [9-PAHSA] and 9-(9Z-octadecenoyloxy)-octadecanoic acid [9-OAHSA], on intact livers in C57BL/6J mice. Materials and methods Short-term in vivo study with intragastric gavage of 13 mg/kg of substances. Morphological, biochemical and high-resolution respirometric assessment of plasma and liver tissue or homogenates thereof. Results The 9-OAHSA-gavaged mice had the highest final total body weight, the lowest free fatty acid circulating levels and the highest plasma activities of both ALT and AST. No significant changes of ambient glycaemia were found, however 9-PAHSA-gavaged mice tended to have lower glycaemia than other animals. Respirometry proved no substance-dependent differences. Discussion and conclusion 9-PAHSA was more metabolically beneficial and less hepatotoxic than 9-OAHSA. Bioenergetic machinery of liver homogenates seemed unaffected at our FAHFA dose.

Membrane Bioreactors for wastewater reuse: Respirometric assessment of biomass activity during a two year survey

Stricter effluent limits, water shortage conditions, land availability requires today even more the needs of advanced wastewater treatments. Attractive solutions come from membrane bioreactors (MBR), Integrated Fixed Film Activated Sludge (IFAS) or combinations (i.e., IFAS-MBRs). One crucial aspect for the applicability of this overall new technology, compared to the conventional activated sludge systems, is the lack of knowledge for design and manage (e.g., kinetic constants, optimal operative conditions etc.). In view of the above frame, the aim of the present study was to assess the kinetic and stoichiometric parameters of bacterial species in MBRs by means of respirometric techniques. Plant configurations, operational conditions and wastewater features (domestic/industrial) were analysed. Four different MBR plants were investigated: i) sequencing batch MBR subjected to a gradual salinity increase; ii) pre-denitrification MBR treating saline wastewater contaminated by hydrocarbons; iii) University of Cape Town (UCT) MBR treating domestic wastewater subjected to a carbon-to-nitrogen (C/N) ratio variation; iiii) UCT- IFAS -MBR treating domestic wastewater. Results show a significant influence on biomass respiratory activity from both plant configurations and operational conditions. The salinity increase severely affected the activity of autotrophic species, while heterotrophic community was mainly influenced by the C/N variation. Moreover, it was observed a specialization in the IFAS-MBR configuration, with the suspended biomass more affine to organic matter, whilst biofilm in the nitrification process. The respirometric analysis confirmed to be an effective tool for the evaluation of the biomass kinetic and stoichiometric parameters. The results of this study can be useful for the application of mathematical models in the design phase and for the monitoring of biomass viability during plant operations.

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 μg of mitochondria indicate that 1.0 μg is sufficient to measure respiration and that 5.0 μg provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Preparation and respirometric assessment of mitochondria isolated from skeletal muscle tissue obtained by percutaneous needle biopsy.

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 μg of mitochondria indicate that 1.0 μg is sufficient to measure respiration and that 5.0 μg provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Effect of storage on the respirometric relationship between substrate utilization and microbial growth

The paper evaluated the impact of substrate storage on the respirometric assessment of process stoichiometry based on oxygen uptake rate (OUR) measurements. Two parallel sequencing batch reactors were operated with pulse feeding of synthetic substrate mixture at a sludge age of 8days and 2days. During the cycle experiments with acetate, 40–45% of acetate was converted to polyhydroxybutyrate, which was partly consumed during each cycle. Respirometric analysis also yielded OUR profiles for the corresponding cyclic operation. A mass balance expression was derived based on oxygen utilization. Oxygen demands calculated on the basis of partial PHB utilization closely matched the experimental values retrieved from OUR profiles within limits of analytical precision. The relative contribution of storage mechanism represented more than 50% of overall oxygen demand. Substrate storage, when totally disregarded or not properly evaluated, was observed to involve an error of around 10% on overall the oxygen demand.

Respirometric assessment of substrate binding by antibiotics in peptone biodegradation

The study evaluated the inhibitory impact of antibiotics on the biodegradation of peptone mixture by an acclimated microbial culture under aerobic conditions. A fill and draw reactor fed with the peptone mixture defined in the ISO 8192 procedure and sustained at steady state at a sludge age of 10 days was used as the biomass pool with a well-defined culture history. Acute inhibition experiments involved running six parallel batch reactors seeded with biomass from the fill and draw reactor and the same peptone mixture together with pulse feeding of 50 mg/L and 200 mg/L of Sulfamethoxazole, Erythromycin and Tetracycline. Substrate utilization was evaluated by observing the respective oxygen uptake rate profiles and compared with a control reactor, which was started with no antibiotic addition. While all available external substrate was removed from solution, addition of antibiotics induced a significant decrease in the amount of oxygen consumed, indicating that a varying fraction of peptone mixture was blocked by the antibiotic and did not participate to the on-going microbial growth mechanism. This observation was also compatible with the concept of the uncompetitive inhibition mechanism, which defines a similar substrate blockage through formation of an enzyme- inhibitor complex.

Respirometric assessment of aerobic sludge stabilization

Aerobic sludge stabilization was assessed respirometrically with the sludge taken from the secondary settling tank of a domestic wastewater treatment facility in Istanbul, Turkey. Zero-order removal rates of 178, 127 and 44 mg/L day were found for Suspended Solids (SS), Volatile Suspended Solids (VSS) and Total Organic Carbon (TOC) at the end of 18 days sludge stabilization, respectively. Significant nutrient release was observed by the mineralized nitrogen and phosphorus from the death and lysis of microorganisms. The model simulations for the batch respirometric assays for initial, 7th and 18th days of the stabilization agree reasonably well with the experimental data. The maximum storage rates (k(sto)) as well as maximum growth rates on stored products (micro(H2)) decrease with increasing stabilization period. Respirometric assays indicated the presence of microorganisms that started to compete with the dominant microorganisms as a result of the stabilization. As such, these findings have significance in terms of the efforts related to the sludge management and application processes.

Nickel inhibition of calcium precipitation by ureolytic mixed microorganisms under batch conditions

The respirometric assessment of the inhibitory impact of Ni(II) on substrate utilization and microbial carbonate precipitation (MCP) by ureolytic mixed microorganisms was investigated with a glucose containing mineral medium under batch conditions over an incubation period of 134 h. The IC 25 was determined as 224 mg Ni(II) L −1 from the BOD values of samples. The interpretation of kinetic data showed that the substrate removal rate fitted a zero-order at the beginning of the incubation period and first-order during the last period, for a range of Ni(II) concentrations between 0 and 512 mg L −1. Increasing Ni(II) concentrations from 0 to 512 mg L −1 reduced the substrate degradation rate constant from 10.8 to 5.3 mg L −1 h −1 for zero-order rate constant ( k 0), and from 0.015 to 0.002 h −1 for first-order rate constant ( k 1). The zero- and first-order reaction rates during incubation period were equalized to the reaction rate of the Monod equation in order to determine the kinetic constants, half saturation concentration ( K S) and maximum substrate removal rate ( R max). BOD removal rate was inhibited accordingly to mixed inhibition model with increasing Ni(II) concentrations during the calcification process. Also, the inhibition of calcium precipitation was observed at a higher Ni(II) concentration because of inhibition of ammonium production in these samples.

Respirometric assessment of biodegradation characteristics of the scientific pitfalls of wastewaters

This paper provides an overview of common problems encountered when using oxygen uptake rate (OUR) measurements for the assessment of wastewater characteristics and process kinetics. Emphasis is placed upon pitfalls that would lead to significant errors. It covers model dependency of the OUR measurements and the need to select appropriate models; interpretation of OUR perturbations as a way to identify new model components and processes; the need for simultaneous observation of relevant model components and multicomponent modelling for appropriate evaluation of OUR measurements; parameter identifiability problems and the effect of active biomass concentration and the endogenous decay rate on model simulation and calibration. Relevant experimental OUR data from previous studies are presented to illustrate and underline common scientific pitfalls.

Respirometric assessment of biodegradation for acrylic fibre-based carpet finishing wastewaters

The paper evaluates biodegradation characteristics of wastewaters generated from acrylic fibre-based carpet processing and manufacturing. It involves detailed characterisation, respirometric modelling and kinetic description of dyeing and softening wastewater streams and the composite effluent. The wastewaters exhibit different COD content and fractionation. The resulting composite effluent has a total COD of 775 mgL(-1), predominantly soluble and with a biodegradable fraction of 86%. In respirometric studies, the OUR profiles can only be calibrated with a dual hydrolysis model with rates significantly slower compared to domestic sewage and other textile plant effluents. Kinetic information derived from the experiments is applied for the conceptual evaluation of the treatability of the composite wastewater using two different continuous-flow activated sludge configurations.

Respirometric Assessment of Primary Sludge Fermentation Products

This study investigates the potential of primary sludge fermentation for the generation of readily biodegradable substrate. Experimental evaluation indicates that uncontrolled fermentation converted 22% of the initial volatile suspended solids in the sludge into soluble biodegradable chemical oxygen demand (COD). More than 85% of the soluble COD generated was associated with the formation of short chain volatile fatty acids (VFAs). The recoverable fraction of the fermented sludge supernatant may potentially increase the biodegradable COD content of the primary effluent by 5%. The VFA composition predominantly involved acetic and propionic acids as reported in the literature. Due to the high VFA content, activated sludge Model No. 1 could not predict the COD fractionation in the primary sludge; activated sludge Model No. 3 provided a better interpretation of the oxygen uptake profile through initial storage of the VFAs in the sludge.

Biodegradation of tannery wastewater using sequencing batch reactor—Respirometric assessment

This investigation proved that respirometry combined with sequencing batch reactor (SBR) could be an effective way for the removal of COD in tannery wastewater. Measurement of oxygen uptake rates (OUR) and corresponding COD uptake rates showed that a 12-h operating cycle was optimum for tannery wastewater. The removal of COD by degradation was stoichiometric with oxygen usage. A plot of OUR values provided a good indication of the biological activity in the reactor. A high OUR value corresponded to the feed period; at the end of the cycle, when the substrate was depleted, the OUR value was low. At a 12-h SBR cycle with a loading rate of 1.9–2.1 kg m −3 d −1, removal of 80–82% COD, 78–80% TKN and 83–99% NH 3-N were achieved. These removal efficiencies were much higher than the conventional aerobic systems. A simple method of COD fractionation was performed from the OUR and COD uptake rate data of the SBR cycle. About 66–70% of the influent COD was found to be readily biodegradable, 10–14% was slowly degradable and 17–21% was non-biodegradable. The oxygen mass transfer coefficient, K L a (19 ± 1.7 h −1) was derived from respirometry. It was observed that with the exception of high organic load at the initial feed the oxygen transfer capacity was in excess of the OUR, and aerobic condition was generally maintained. Simultaneous nitrification–denitrification was observed in the SBR during the feed period as proved by mass balance.

Respirometric assessment of residual organic matter for domestic sewage

A new method is developed for the respirometric assessment of the soluble and particulate COD components in wastewater. The method relies on evaluating the area under the oxygen uptake rate (OUR) curve, excluding the fraction associated with endogenous respiration, generated for a wastewater sample as the equivalent of its biodegradable COD content. The method compares well with a previously recommended procedure for the calculation of the soluble inert COD. It also enables the computation of the soluble residual microbial products likely to be generated during the course of biological processes.

Respirometric assessment of storage yield for different substrates

A new procedure has been defined for the respirometric assessment of bacterial storage yield as defined in the Activated Sludge Model No.3. The procedure was used to determine the storage yield, Y(STO), associated with acetate, glucose and domestic sewage, together with mixtures of acetate/glucose and acetate/domestic sewage at different initial F/M ratios. Y(STO) was calculated as 0.78 gCOD(gCOD)(-1) for acetate, 0.87 gCOD(gCOD)(-1) for glucose and 0.96 gCOD(gCOD)(-1) for domestic sewage. The Y(STO) of substrate mixtures was found to reflect the characteristics of the dominant fraction in the mixture.