Effect of high substrate concentration and cell feedback on kinetic behavior of heterogeneous populations in completely mixed systems

Growth kinetics of heterogeneous populations of sewage origin were studied in completely mixed reactors of the once-through type at a high concentration of incoming substrate, 3000 mg/l glucose, and in systems employing cell feedback or sludge recycle at an incoming substrate concentration of 1000 mg/1 glucose. The recycle flow rate employed was 25% of the incoming feed flow, and the concentration of cells in the recycle was maintained as closely as possible at 150% of the cell concentration in the reactor. Studies were made at various dilution rates. Throughout these studies, batch experiments using cells grown at the various dilution rates were made to determine k_s and μ_m values. As in previous studios using heterogeneous populations, the relationship between specific growth rates μ and substrate concentration S was represented better by the Monod equation than by any other which was tested. The growth “constants” μ_m, k_s, and Y were found to fall in the same general range as those determined in previous studies in once-through systems operated at 1000 mg/l glucose. It was observed that cell recycle, even at the relatively low concentration factor employed in these studies, greatly enhanced the flocculating and settling characteristics of the cells.     

Fermentation profiles in bioconversions

Biotechnology Letters - Any complex substrate biomass can be characterised by its “fermentation profile”, i.e. a complex substrate biomass, in the presence of an ubiquitous microbial...     

Transient Response of Continuously Cultured Heterogeneous Populations to Changes in Temperature

Completely mixed, once-through continuous culture systems of heterogeneous microbial populations of sewage origin were systematically examined for response to changes in reactor temperature. Systems were operated at two dilution rates of 0.125 and 0.25 per h. "Steady state" conditions of the systems were assessed with the reactors operating at 25 C. From this base line, temperature was decreased to as low as 8 C and increased to as high as 57.5 C. Response was assessed in the ensuing transient phase as the system approached a new "steady state." The response was measured by changes in amount and type of carbon source in the reactor effluent as determined by the chemical oxygen demand test, the anthrone test, and gas chromatography. Biological solids concentration and cell composition (protein, carbohydrate, ribonucleic acid and deoxyribonucleic acid) were also determined. These systems responded more favorably to increases than to decreases in temperature. Regardless of the direction of change, the system with the lowest dilution rate (D = 0.125 per h) responded more successfully; i.e., there was less leakage of carbon source in the effluent and less dilute-out of cells during the transient phase.     

Physiological state of a microbial community in a biomass recycle reactor

The transition in physiological state was investigated between a carbon-limited chemostat population and microbes growing very slowly in a biomass recycle reactor. The mixed microbial population was metabolizing a mixture of biopolymers and linear alkylbenzene sulfonate, formulated to represent the organic load in graywater. Biomass increased 30-fold during the first 14 days after a shift from chemostat to biomass recycle mode. The ratios of ATP and RNA to cell protein decreased over the first days but then remained constant. The specific rate of CO₂ production by microbes in the reactor decreased 6-fold within 24 h after the shift, and respiratory potentials declined 2–3 fold during the first 7 days. Whereas chemostat cultures used equal proportions of organic carbon substrate for catabolism and anabolism, the proportion of organic substrate oxidized to CO₂ rose from 62 to 82% over the first 8 days in a biomass recycle reactor, and eventually reached 100% as this reactor population exhibited no net growth. Biomass recycle populations removed from the system and subjected to a nutritional shift-up did not immediately initiate exponential growth. The physiological state of cells in the biomass recycle reactor may be distinct from those grown in batch or continuous culture, or from starved cells. Received 02 June 1997/ Accepted in revised form 20 February 1998     

Biomass Carbon Measurements and Substrate Utilization Patterns of Microbial Populations from Soils Amended with Cadmium, Copper, or Zinc

Samples of a sandy loam soil taken from a long-term liming experiment in southeast England were amended with solutions of metal sulfate salts. Soils with a range of pHs were amended to contain Cu, Cd, or Zn at concentrations around the maximum permissible values for these metals in agricultural land receiving sewage sludge. After a 3-year equilibration period, the microbial biomass was determined by the fumigation-extraction technique. These results were compared with data from substrate utilization patterns of microbial populations extracted by using a weak salt solution. There was no reduction in microbial biomass due to pH or metal treatment in any of the soils except the Cu treatment. Principal-component analysis of the respiration patterns in Biolog plates demonstrated effects of both pH and metal treatment on the extracted microbial population which were independent of gross biomass size. pH and soil amendments with Cu and Zn were found to reduce the metabolic potential of the extracted soil microbial population.     

Two-enzyme active transport in vitro with ph induced asymmetrical functional structures.: III. Discussions based on numerical treatments of the time-dependent evolutions

Three complementary models have been considered in which pH gradients (step function. linear pH or linear H^?) impose asymmetry on a two-enzyme mixture. If the “combined pH dependences” of enzymes is pro-asymmetrical, the pH gradient induces an asymmetrical distribution of potential activities (“latent” asymmetry of functional structure). When substrate is added, “developed” asymmetry of effective activities appears which results in “substrate space wave” and pumping when the catalysed reaction couple is “inversible”. It is shown that only one steady state exists for a given boundary condition and is attained when the “combined effective activity” of enzymes is nil: the stationary flux with symmetrical boundaries or the stationary load with moving boundaries is proportional to “effective global activities” of enzymes. “Equivalent square models” could be proposed that would be able to describe “functional” or “permanent” structure pumps as well. These models belong to the thermodynamic branch and the asymmetrical “space wave” substrate concentration profiles obtained must be distinguished from dissipative structures. It appears that such primary active transport pumps are chemical equivalents of heat pumps.     

Operating parameters' effects on the outcome of pure and simple competition between two populations in configurations of two interconnected chemostats

It is known from the literature that two microbial populations competing purely and simply for a common substrate in a spatially inhomogeneous environment may under certain conditions coexist in a steady state. This paper studies pure and simple competition between two microbial species in three alternate configurations of two interconnected ideal chemostats and focuses on the effects of the operating parameters-dilution rate, substrate concentration in the feed to the vessels, recycle ratio, and volume ratio of the two vessels, splitting ratio of the external feed to the chemostats-on the coexistence of the two competitors. It is shown that the coexistence steady state is practically feasible in the sense that it occurs in a finite domain of the operating parameters space. Theoretical and numerical results are presented, some of them in the form of operating diagrams projected on the two-dimensional subspace. A comparison of the three possible configurations is offered.     

Nitrogen metabolism in extended aeration processes operated with and without hydrolytic pretreatment of portions of the sludge

Nitrifying characteristics were compared for an extended aeration (total cell recycle) process and one employing an engineering modification of the process utilizing a “hydrolytic assist” to aid biological autodigestion. Laboratory pilot plants were run over a period of years, and it was found that the recently recommended “hydrolytic assist” did not militate against production of a highly nitrified effluent. Under this mode of operation, the effluent was as nitrified as the effluent from the normal extended aeration process. It was also found that the modified process rapidly recovered its nitrifying capability after a period of deprivation of excess ammonia nitrogen. Throughout the period of operation, substrate removal efficiency remained high.     

Eco-physiological Characterization of Soil Bacterial Populations in Different States of Growth

The method based on characterization of microbial populations in terms of their growth rate in agar plates has been used for testing the prediction of the theory of r- and K-selection in a microbial community from a tropical soil. Conditions which could lead bacterial populations to grow exponentially or to enter into a stationary phase were obtained by growing soil microbial populations in a chemostat and in a chemostat with recycle, respectively. Significant differences in population distribution patterns were observed by comparing results from the two growth systems. When soil community was grown in a chemostat and subjected specifically to well-defined r- and K-conditions, stable associations of organisms with r- and K-type characteristics developed as a consequence of environmental pressure. In contrast, when cultivated in chemostat with recycle under the same r- and K-conditions imposed on chemostat cultures, distribution patterns of r- and K-selected populations appeared very little affected by changes in substrate availability.     

Steady-state model for activated sludge with constant recycle sludge concentration

In a previous report it was concluded that steady-state operation of completely mixed reactors for growth of heterogeneous microbial populations, i.e., activated sludge processes, was extremely difficult to attain if maintenance of a constant sludge recycle ratio, c, was required, and equations were devised in which the concentration of cells in the recycle, x_R, rather than the recycle ratio, was constant. In this report the equations are developed and computational analysis shows the effect on substrate and cell concentrations in the reactor of operational variables such as inflowing feed concentration, hydraulic recycle ratio, recycle sludge concentration, dilution rate, and the biological “constants” μ_m, k_s, and Y. The stabilizing effect of operating with constant x_R on the dilute-out pattern is shown.     

Simultaneous single-cell protein production and COD removal with characterization of residual protein and intermediate metabolites during whey fermentation by K. marxianus

Cheese whey fermentation with Kluyveromyces marxianus was carried out at 40 °C and pH 3.5 to examine simultaneous single-cell protein production and chemical oxygen demand (COD) removal, determine the fate of soluble whey protein and characterize intermediate metabolites. After 36 h of batch fermentation, the biomass concentration increased from 2.0 to 6.0 g/L with 55 % COD reduction (including protein), whereas soluble whey protein concentration decreased from 5.6 to 4.1 g/L. It was confirmed through electrophoresis (SDS-PAGE) that the fermented whey protein was different from native whey protein. HPLC and GC–MS analysis revealed a change in composition of organic compounds post-fermentation. High inoculum concentration in batch fermentation resulted in an increase in biomass concentration from 10.3 to 15.9 g/L with 80 % COD reduction (including protein) within 36 h with residual protein concentration of 4.5 g/L. In third batch fermentation, the biomass concentration increased from 7.3 to 12.4 g/L with 71 % of COD removal and residual protein concentration of 4.3 g/L after 22 h. After 22 h, the batch process was shifted to a continuous process with cell recycle, and the steady state was achieved after another 60 h with biomass yield of 0.19 g biomass/g lactose and productivity of 0.26 g/L h. COD removal efficiency was 78–79 % with residual protein concentration of 3.8–4.2 g/L. The aerobic continuous fermentation process with cell recycle could be applied to single-cell protein production with substantial COD removal at low pH and high temperature from cheese whey.     

蚂蚁筑巢对西双版纳热带森林土壤微生物生物量碳及熵的影响

蚂蚁筑巢能够改变热带森林土壤理化环境,从而对土壤微生物生物量碳及熵的时空动态产生重要影响.本研究以西双版纳高檐蒲桃热带森林群落为对象,采用氯仿熏蒸法对蚂蚁巢地和非巢地土壤微生物生物量碳及熵时空动态进行测定.结果表明: 1)蚁巢地平均微生物生物量碳及熵(1.95 g·kg^-1,6.8%)显著高于非巢穴(1.76 g·kg^-1,5.1%);蚁巢地和非蚁巢地土壤微生物生物量碳呈单峰型时间变化趋势,而土壤微生物熵呈“V”型变化格局.2)蚁巢地和非巢地土壤微生物生物量碳及熵均具有明显的垂直变化:微生物生物量碳随土层加深显著降低,微生物熵则沿土层加深显著升高,但蚁巢微生物生物量碳及熵的垂直变化较非巢穴显著. 3)蚂蚁筑巢引起了巢内水分和温度的显著改变,进而影响土壤微生物生物量碳及熵的时空动态.土壤水分分别解释微生物生物量碳及熵的66%～83%和54%～69%,而土壤温度分别解释土壤微生物生物量碳及熵的71%～86%和67%～76%. 4)蚂蚁筑巢引起土壤理化性质变化对土壤微生物生物量碳和熵产生重要影响.蚁巢土壤微生物生物量碳与土壤有机碳、温度、全氮、含水率呈极显著正相关,与容重、硝态氮,水解氮呈显著正相关,与土壤pH呈极显著负相关;除土壤微生物熵与pH呈显著正相关外,与其他土壤理化指标均呈显著负相关.土壤总有机碳、全氮和温度对微生物生物量碳的贡献最大,而土壤总有机碳和全氮对微生物熵的负作用最小.因此,蚂蚁筑巢能够显著改变微生境(如土壤水分与温度)及土壤理化性质(如总有机碳及全氮),进而调控热带森林土壤微生物生物量碳及熵的时空动态.     

Biological kinetic behaviors and operational performances in aerated and oxygenated systems

Biological kinetic behaviors of the oxygenated and aerated activated sludge process were studied and compared in both once-through and constant sludge recycle systems. The models derived by Herbert, Elsworth, and Telling [J. Gen. Microbiol., 14 , 601 (1956)] and Ramanathan and Gaudy [Biotechnol. Bioeng., 11 , 207 (1969)] were used for the studies of once-through and constant sludge recycle systems, respectively. Soft drink waste water was used for the growth limiting substrate. Temperature was controlled within 30 ± 2°C. The influent substrate concentration was maintained at 1,000 mg/liter. The experiments were conducted at various dilution rates (from \documentclass{article}\pagestyle{empty}\begin{document}$ \frac{1}{9} $\end{document} to 1/1.0 hr^?1), and recycle solids concentration values (from 5,000 to 10,000 mg/liter), with hydraulic recycle ratio, α, at 0.3. Biological kinetic constants were evaluated and compared. It was found that these constants were different for the aerated and oxygenated systems within a certain range of dilution rates studied. The critical dilution rates for diluting out effluent chemical oxygen demand (COD) occurred at 0.1 and 0.2 hr^?1in the once-through operation, and 0.2 and 0.4 hr^?1in the sludge recycle operation for aerated and oxygenated systems, respectively. Observed sludge yield values and specific growth rate were varied with the type of aeration and with and without constant sludge recycle concentration applied. Sludge carbohydrates and proteins content in the oxygenation system (cell recycle) were 10.1–21.6% and 35.6–52.2%. Sludge volume index in the air and oxygenation systems varied from 41.4 to 354 and 31.9 to 58.5, respectively.     

Identification of some of the major groups of bacteria in efficient and nonefficient biological phosphorus removal activated sludge systems.

To investigate the bacteria that are important to phosphorus (P) removal in activated sludge, microbial populations were analyzed during the operation of a laboratory-scale reactor with various P removal performances. The bacterial population structure, analyzed by fluorescence in situ hybridization (FISH) with oligonucleotides probes complementary to regions of the 16S and 23S rRNAs, was associated with the P removal performance of the reactor. At one stage of the reactor operation, chemical characterization revealed that extremely poor P removal was occurring. However, like in typical P-removing sludges, complete anaerobic uptake of the carbon substrate occurred. Bacteria inhibiting P removal overwhelmed the reactor, and according to FISH, bacteria of the beta subclass of the class Proteobacteria other than beta-1 or beta-2 were dominant in the sludge (58% of the population). Changes made to the operation of the reactor led to the development of a biomass population with an extremely good P removal capacity. The biochemical transformations observed in this sludge were characteristic of typical P-removing activated sludge. The microbial population analysis of the P-removing sludge indicated that bacteria of the beta-2 subclass of the class Proteobacteria and actinobacteria were dominant (55 and 35%, respectively), therefore implicating bacteria from these groups in high-performance P removal. The changes in operation that led to the improved performance of the reactor included allowing the pH to rise during the anaerobic period, which promoted anaerobic phosphate release and possibly caused selection against non-phosphate-removing bacteria.     

Four years of litter input manipulation changes soil microbial characteristics in a temperate mixed forest

Aboveground litter not only is an important source of nutrients to soil microbes but also regulates the microclimate in topsoil. How the changes in aboveground litter quantity would affect the microbial biogeochemical cycles is still unclear. Here we conducted a litter input manipulation experiment in a temperate mixed forest to investigate how different amounts of litter input affect soil organic carbon (SOC) and soil respiration via their regulation on soil microbes. We found that although neither SOC stock nor soil CO₂ efflux was affected by litter manipulation, soil microbial characteristics had responded after four years of litter addition or removal treatments. Microbial biomass carbon (MBC) in the O horizon was higher in litter addition plots than in litter removal plots as a result of the changed availability of labile C under litter treatments. Both double litter and no litter treatments changed microbial compositions, which was probably due to the increased soil pH in no litter treatment and the increased labile C in double litter treatment. The null change in soil respiration could be attributed to the offset between the negative effect of decreased substrate and the positive effect of increased temperature on soil respiration in litter removal plots. Due to the important role of soil microbes in carbon cycling, the altered microbial properties under litter manipulation treatments suggested the inevitable changes in biogeochemical cycling in the long run and call for long-term studies on SOC dynamics in the future.

     

The transient response to pH and temperature shock loading of fermentation systems

Recent findings in the author's laboratories relative to the determination of the mode of response of continuously cultured heterogeneous populations to step changes in pH and in temperature are reviewed and discussed in light of possible effects of specific growth rate, μ, on the severity of substrate leakage in the effluent during the transient phase.     

Effect of excess ethanol on the growth of yeasts of the genus Candida during continuous cultivation

The effect of flow rates and a specific ethanol load on the growth of Candida utilis and Candida krusei was studied in the process of one-step and three-step cultivation. The productive capacity of fermenters and the economic coefficient of yeast biomass production were shown to depend on the ability of microbial populations to assimilate a certain quantity of a carbon substrate per unit time. When a specific ethanol load exceeds the optimal one, the respiratory activity of a population and the economic coefficient of growth fall down whereas the accumulation of metabolites in the cultural broth increases. The steady state of biomass can be maintained in the process of continuous cultivation by inhibiting the yeast growth with an excess of ethanol.     

Diffusion and reaction within porous packing media: a phenomenological model

A phenomenological model has been developed to describe biomass distribution and substrate depletion in porous diatomaceous earth (DE) pellets colonized by Pseudomonas aeruginosa. The essential features of the model are diffusion, attachment and detachment to/from pore walls of the biomass, diffusion of substrate within the pellet, and external mass transfer of both substrate and biomass in the bulk fluid of a packed bed containing the pellets. A bench-scale reactor filled with DE pellets was inoculated with P. aeruginosa and operated in plug flow without recycle using a feed containing glucose as the limiting nutrient. Steady-state effluent glucose concentrations were measured at various residence times, and biomass distribution within the pellet was measured at the lowest residence time. In the model, microorganism/substrate kinetics and mass transfer characteristics were predicted from the literature. Only the attachment and detachment parameters were treated as unknowns, and were determined by fitting biomass distribution data within the pellets to the mathematical model. The rate-limiting step in substrate conversion was determined to be internal mass transfer resistance; external mass transfer resistance and microbial kinetic limitations were found to be nearly negligible. Only the outer 5% of the pellets contributed to substrate conversion. (c) 1993 Wiley & Sons, Inc.     

Experimental studies on a kinetic model for design and operation of activated sludge processes

Previous experimentation in our laboratory has shown that the classical theory developed for continuous growth of pure cultures in completely mixed aerobic systems in which the recycle cell concentration factor, c (where c = X_R/X), is a selectable system constant, did not provide a suitable model for the heterogeneous (natural) populations of the activated sludge process. Another model was derived in which the recycle cell concentration, X_R was employed as a system constant instead of c, and computational analysis was performed. Laboratory pilot plant experimentation was undertaken in order to determine whether a “steady state” in aerator biological solids concentration, X?, and substrate concentration, S?, could be approached under this mode of operation. Studies were performed at various organic feed concentrations holding dilution rate, D, at 0.125 hr^?1, hydraulic recycle ratio, α, at 0.25, and X^R at 10,000 mg/liter. Also, values of maximum specific growth rate, μ_max, and saturation constant, K_s were determined. It was found that the model approached the steady state condition with heterogeneous populations more closely than did the classical model, and the high degree of treatment efficiency predicted by the model was demonstrated experimentally.     

Dynamic and steady state studies of phenol biodegradation in pure and mixed cultures.

The microbial degradation of phenol by pure and mixed cultures of Pseudomonas putida was studied in batch, phenol-stat, and continuous culture systems. In the continuous culture runs, both steady state and transient experiments were performed. From these experiments, a model for the kinetic behavior of the organisms was evolved and an analysis performed on the stability and dynamic behavior of pure and mixed cultures. The results indicate that it should be possible to achieve phenol removal from wastewaters down to levels of 1-2 ppm in a single state system. However, because of the effect of substrate inhibition on kinetic behavior of the microorganisms, long lasting transients can occur. The transient behavior of such systems cannot be solely determined from mumax or Ks parameters, but must include a consideration of the transient size and response characteristic of the organism.