Effect of temperature on bacterial species diversity in thermophilic solid-waste composting.

Continuously thermophilic composting was examined with a 4.5-liter reactor placed in an incubator maintained at representative temperatures. Feed was a mixture of dried table scraps and shredded newspaper wetted to 55% moisture. One run at 49 degrees C (run A) employed a 1:4 feed-to-compost ratio, while the other runs used a 10:1 ratio and were incubated at 50, 55, 60, or 65 degrees C. Due to self-heating, internal temperatures of the composting mass were 0 to 7 degrees C hotter than the incubator. Two full-scale composting plants (at Altoona, Pa., and Leicester, England) were also examined. Plate counts per gram (dry weight) on Trypticase soy broth (BBL Microbiology Systems) with 2% agar ranged from 0.7 X 10(9) to 5.3 X 10(9) for laboratory composting and 0.02 X 10(9) to 7.4 X 10(9) for field composting. Fifteen taxa were isolated, including 10 of genus Bacillus, which dominated all samples except that from run A. Species diversity decreased markedly in laboratory composting at 60 degrees C and above, but was similar for the three runs incubated at 49, 50, and 55 degrees C. The maximum desirable composting temperature based on species diversity is thus 60 degrees C, the same as that previously recommended based on measures of the rate of decomposition.     

Biokinetic analyses of adaptation and succession: microbial activity in composting municipal sewage sludge.

The interactions between temperature and the microbial communities in composting municipal sewage sludge were studied to determine the optimal temperature range for efficient decomposition (stabilization) of the sludge. Information concerning thermophilic successions in such communities was also obtained. Samples were taken from several different temperature areas in a production-scale composting pile throughout the 19-day processing run. Optimum temperatures for microbial activity, determined as the rate of [14C]acetate incorporation into microbial lipids, were determined for each sample. Biomass was determined from the lipid phosphate content of the sample. Maximal activities were generally found in samples coming from lower-temperature areas (25 to 45 degrees C), whereas samples from high temperatures (55 to 74 degrees C) usually had relatively little activity. The temperature giving the optimum activity in samples incubated at a variety of temperatures during the assay tended to increase as the composting time progressed, but never exceeded about 50 degrees C. Many of these temperature response curves were similar in nature to curves reported for purified enzyme systems and pure cultures of bacteria. Comparisons of the apparent energies of activation calculated for different temperature ranges over time also indicated that the overall community was better adapted to higher temperatures during the latter part of the composting run. It was also found that the relationship between the apparent energies of activation and the apparent energies of inactivation (apparent heats of denaturation) consistently changed with sample temperature throughout the composting run, suggesting that the microbial communities from hotter samples were better adapted to high temperatures than those from cooler samples, and vice versa.(ABSTRACT TRUNCATED AT 250 WORDS)     

Biokinetic analyses of adaptation and succession: microbial activity in composting municipal sewage sludge

The interactions between temperature and the microbial communities in composting municipal sewage sludge were studied to determine the optimal temperature range for efficient decomposition (stabilization) of the sludge. Information concerning thermophilic successions in such communities was also obtained. Samples were taken from several different temperature areas in a production-scale composting pile throughout the 19-day processing run. Optimum temperatures for microbial activity, determined as the rate of [14C]acetate incorporation into microbial lipids, were determined for each sample. Biomass was determined from the lipid phosphate content of the sample. Maximal activities were generally found in samples coming from lower-temperature areas (25 to 45 degrees C), whereas samples from high temperatures (55 to 74 degrees C) usually had relatively little activity. The temperature giving the optimum activity in samples incubated at a variety of temperatures during the assay tended to increase as the composting time progressed, but never exceeded about 50 degrees C. Many of these temperature response curves were similar in nature to curves reported for purified enzyme systems and pure cultures of bacteria. Comparisons of the apparent energies of activation calculated for different temperature ranges over time also indicated that the overall community was better adapted to higher temperatures during the latter part of the composting run. It was also found that the relationship between the apparent energies of activation and the apparent energies of inactivation (apparent heats of denaturation) consistently changed with sample temperature throughout the composting run, suggesting that the microbial communities from hotter samples were better adapted to high temperatures than those from cooler samples, and vice versa.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the actinomycetal communities during continuous thermophilic composting as revealed by denaturing gradient gel electrophoresis and quantitative PCR

Actinomycetes degrade cellulose and solubilize lignin during composting. Changes in the diversity of the actinomycetal communities and the 16S rDNA copy numbers of actinomycetes were monitored by denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR), respectively, during continuous thermophilic composting (CTC) and traditional composting (TC). qPCR indicated that the copy numbers from the CTC samples were 25-80% higher than those from the TC samples during similar phases of active composting and they were lower than 3×10(9) gene copies/g (dry weight) in the mature compost from both runs. DGGE showed a more diverse actinomycetal community in the CTC than in TC, averaging 16 bands as compared to 12 bands, at the post peak temperature phase. The study suggested that temperatures higher than 50 °C in CTC benefited the growth of actinomycetes.     

Acetate Degradation at 70 degrees C in Upflow Anaerobic Sludge Blanket Reactors and Temperature Response of Granules Grown at 70 degrees C

Anaerobic acetate degradation at 70 degrees C and at 55 degrees C (as a reference) was studied by running laboratory upflow anaerobic sludge blanket (UASB) reactors inoculated with mesophilic granular sludge. In UASB reactors fed with acetate-containing media (3 g of chemical oxygen demand [COD] per liter, corresponding to 47 mM acetate) approximately 50 days was needed at 70 degrees C and less than 15 days was needed at 55 degrees C to achieve an effluent COD of 500 to 700 mg/liter. In the UASB reactors at both 70 and 55 degrees C up to 90% of the COD was removed. Batch assays showed that sludges from two 70 degrees C UASB reactors, one run at a low effluent acetate concentration and the other run at a high effluent acetate concentration, exhibited slightly different responses to temperatures in the range from 37 to 70 degrees C. Both 70 degrees C sludges, as well as the 55 degrees C sludge, produced methane at temperatures of 37 to 73 degrees C. The 55 degrees C sludge exhibited shorter lag phases than the 70 degrees C sludges and higher specific methane production rates between 37 and 65 degrees C.     

Effects of oxygen on aerobic solid-state biodegradation kinetics

Oxygen is a critical control variable for composting and other solid-state biodegradation processes. In this study we examined the effect of varying oxygen concentrations (1%, 4%, and 21% O2 (v/v)) on biodegradation kinetics under different substrate (sewage sludge and synthetic food waste), temperature (35, 45, 55, and 65 degrees C), and moisture (36-60% H2O) conditions. Three forms of a saturation or Monod-type model and one form of an exponential model were evaluated against data from extensive experiments under carefully controlled environmental conditions. The exponential model performed well at temperatures from 35 to 55 degrees C but had problems at higher temperatures. The Monod-type models yielded the best fit based on R2 values. Multiple linear regression was used to express the oxygen half-saturation coefficient as a function of temperature and moisture. For a modified one-parameter saturation model the half-saturation coefficient varied from -0.67% to 1.74% v/v O2 under the range of conditions typical of composting systems. While the positive correlation of biodegradation rate with oxygen concentration reported by previous researchers held true for temperatures below 55 degrees C, an inverse relationship was found at 65 degrees C. Although this study did not directly examine anaerobic conditions, the results under microaerophilic conditions suggest oxygen may not offer kinetic advantages for extreme thermophilic biodegradation processes.     

Intracellular avian type X collagen in situ and determination of its thermal stability using a conformation-dependent monoclonal antibody

The thermal stability of the helical domain of intracellular and matrix-associated type X collagen was examined in situ within the hypertrophic region of embryonic chick vertebral cartilages. For this we employed indirect immunofluorescence histochemistry of unfixed tissue sections reacted at progressively higher temperatures (Linsenmayer et al., J cell biol 99 (1984) 1405) with a conformation-dependent monoclonal antibody (X-AC9) (Schmid & Linsenmayer, J cell biol 100 (1985) 598). The hypertrophic chondrocytes which had most recently initiated synthesis of type X did not immediately secrete it, but instead retained it intracellularly within cytoplasmic organelles. This allowed for clear visualization of the intracellular type X. Within the pool of intracellular type X collagen, the epitope recognized by the antibody was stable up to 55 degrees C, but was destroyed at 60 degrees C. This is 5-10 degrees C higher than the thermal stability of the epitope when the molecule is in neutral solution (as determined by competition ELISA). The matrix-associated type X collagen is stable at least to 65-67.5 degrees C. We conclude that in situ the stability of the collagen helix in its normal intracellular environment is considerably greater than might be predicted from measurements made on molecules in solution.     

Physical and Chemical Correlates of Microbial Activity and Biomass in Composting Municipal Sewage Sludge

Various physical and chemical parameters were monitored to evaluate their influence on the microbial communities present in composting municipal sewage sludge. Temperature, moisture content, depth, pH, protein content, total nitrogen, total carbon, lipid phosphate biomass, and the rates of microbial incorporation of substrates into lipids were measured at several times throughout the 17- to 19-day composting runs. Temperature was found to have the most consistent and dramatic effect on microbial activity and biomass. When temperatures exceeded 55 to 60°C, microbial activity fell dramatically, usually by more than 1 order of magnitude. Microbial activity was generally greatest in samples taken from the 35 to 50°C areas of the composting piles. Changes in the composition of the compost over time included increased pH, increased protein content, and decreased total organic content. The changes in these parameters appeared to reflect the microbial activity and biomass present. The results of this study indicate that the rate of composting may best be optimized by controlling the composting temperatures, provided that the other parameters fall within reasonable limits in the starting material.     

Anaerobic treatment of poultry mortality in a temperature-phased leachbed-UASB system

Anaerobic digestion has been proposed as an alternative to the conventional disposal methods of burial, incineration, rendering and aerobic composting. A temperature-phased system consisting of one UASB (at 55 degrees C) and three leach-bed reactors (at ambient temperatures) was tested for its efficiencies in treating poultry mortality. The thermophilic UASB was difficult to start-up. It also showed signs of inhibited methanogenesis. Chemical parameters such as long chain fatty acids, volatile fatty acids and ammonia concentrations were all very high for the thermophilic UASB. Lowering its temperature to 35 degrees C enhanced its stability and improved its performances. Lowering the pH of the 55 degrees C UASB also improved its chemical oxygen demand (COD) reduction efficiency as well as its methane production rate. The results were compared to that of another similar system where the UASB reactor was maintained at 35 degrees C instead of at 55 degrees C.     

Growth of Listeria monocytogenes at refrigeration temperatures

The growth of three strains of Listeria monocytogenes at refrigeration temperatures (-0.5 to 9.3 degrees C) in chicken broth and/or UHT milk was determined using a rocking temperature gradient incubator. Minimum growth temperatures ranged from -0.1 to -0.4 degree C for the three strains. Lag times of 1-3 d and 3 to greater than 34 d were observed with incubation at 5 and 0 degrees C respectively. Corresponding generation times ranged from 13-24 h at 5 degrees C and 62-131 h at 0 degree C. The type of culture medium had an influence on both the rate and extent of growth. Incubation of cultures at 4 degrees C before inoculation caused a marked reduction in the lag time when compared with cultures which had been previously incubated at 30 degrees C.     

Trehalose uptake through P2X7 purinergic channels provides dehydration protection

The tetra-anionic form of ATP (ATP4-) is known to induce monovalent and divalent ion fluxes in cells that express purinergic P2X7 receptors and with sustained application of ATP it has been shown that dyes as large as 831 Da can permeate the cell membrane. The current study explores the kinetics of loading alpha,alpha-trehalose (342 Da) into ATP stimulated J774.A1 cells, which are known to express the purinergic P2X7 receptor. Cells that were incubated at 37 degrees C in a 50 mM phosphate buffer (pH 7.0) containing 225 mM trehalose and 5 mM ATP, were shown to load trehalose linearly over time. Concentrations of approximately 50 mM were reached within 90 min of incubation. Cells incubated in the same solution at 4 degrees C loaded minimally, consistent with the inactivity of the receptor at low temperatures. However, extended incubation at 37 degrees C (>60 min) resulted in zero next-day survival, with adverse effects appearing even with incubation periods as short as 30 min. By using a two-step protocol with a short time period at 37 degrees C to allow pore formation, followed by an extended loading period on ice, cells could be loaded with up to 50 mM trehalose while maintaining good next day recovery (49 +/- 12% by Trypan blue exclusion, 56 +/- 20% by alamarBlue assay). Cells porated by this method and allowed an overnight recovery period exhibited improved dehydration tolerance suggesting a role for ATP poration in the anhydrous preservation of cells.     

Biology of Cerotoma arcuatus (Coleoptera: Chrysomelidae) and field validation of a laboratory model for temperature requirements

Cerotoma arcuatus Olivier is a polyphagous pest of legumes [soybean, Glycine max (L.); dry beans, Phaseolus vulgaris (L.); and cowpeas, Vigna unguiculata (L.)], all of which are considered important protein sources for humans and domestic animals. Studies on the biology and temperature requirements of C. arcuatus were made under laboratory and field (cage) conditions. In the laboratory, insects were reared on soybean plants in incubators held at 18, 20, 22, 25, 28, 30, or 32 degrees C, 70 +/- 10% RH, and a photoperiod of 14:10 (L:D) h. A degree-days (DD) model developed based on the incubator data were validated in the field based on air and soil temperatures. The duration of the egg, larva-to-adult, and egg-to-adult period was inversely correlated with the temperature within the range of 18-32 degrees C, with the highest viability found from 20 to 30 degrees C. The temperature threshold for development and the thermal constant for the egg phase and the larva-to-adult periods were 13.6 degrees C and 106.7 DD, 8.3 degrees C and 399.4 DD, and 10.7 degrees C and 489.0 DD, respectively. The DD model for the egg-to-adult period, calculated using constant temperatures in the laboratory, was found to be valid for populations of C. arcuatus in the field, based on the fluctuating air and soil temperatures, although air temperatures provided more precise predictions. These data provide support for the rational control of this pest through population predictions based on their temperature requirements.     

Effects of Temperature on Methanogenesis in a Thermophilic (58 degrees C) Anaerobic Digestor

The short-term effects of temperature on methanogenesis from acetate or CO(2) in a thermophilic (58 degrees C) anaerobic digestor were studied by incubating digestor sludge at different temperatures with C-labeled methane precursors (CH(3)COO or CO(2)). During a period when Methanosarcina sp. was numerous in the sludge, methanogenesis from acetate was optimal at 55 to 60 degrees C and was completely inhibited at 65 degrees C. A Methanosarcina culture isolated from the digestor grew optimally on acetate at 55 to 58 degrees C and did not grow or produce methane at 65 degrees C. An accidental shift of digestor temperature from 58 to 64 degrees C during this period caused a sharp decrease in gas production and a large increase in acetate concentration within 24 h, indicating that the aceticlastic methanogens in the digestor were the population most susceptible to this temperature increase. During a later period when Methanothrix sp. was numerous in the digestor, methanogenesis from CH(3)COO was optimal at 65 degrees C and completely inhibited at 75 degrees C. A partially purified Methanothrix enrichment culture derived from the digestor had a maximum growth temperature near 70 degrees C. Methanogenesis from CO(2) in the sludge was optimal at 65 degrees C and still proceeded at 75 degrees C. A CO(2)-reducing Methanobacterium sp. isolated from the digestor was capable of methanogenesis at 75 degrees C. During the period when Methanothix sp. was apparently dominant, sludge incubated for 24 h at 65 degrees C produced more methane than sludge incubated at 60 degrees C, and no acetate accumulated at 65 degrees C. Methanogenesis was severely inhibited in sludge incubated at 70 degrees C, but since neither acetate nor H(2) accumulated, production of these methanogenic substrates by fermentative bacteria was probably the most temperature-sensitive process. Thus, there was a correlation between digestor performance at different temperatures and responses to temperature by cultures of methanogens believed to play important roles in the digestor.     

Viability of Rhodococcus equi and Parascaris equorum Eggs Exposed to High Temperatures

There is great concern about the potential pathogen contamination of horse manure compost spread in the same fields horses graze in. To ensure that pathogen destruction occurs, temperatures need to be sufficiently high during composting. Here, we investigated the survival rate of two marker organisms, Rhodococcus equi and Parascaris equorum eggs, exposed to temperatures potentially encountered during horse manure composting. Our results show that the time required to achieve a 1 log10 reduction in R. equi population (D-value) are 17.1 h (±1.47) at 45°C, 8.6 h (±0.28) at 50°C, 2.9 h (±0.04) at 55°C and 0.7 h (±0.04) at 60°C. For P. equorum eggs we show that at 45 and 50°C, 2 log10 reduction of viability is reached between 8 and 24 h of incubation and that it takes less than 2 h at 55 and 60°C to achieve a viability reduction of 2 log10. These results are useful for identifying composting conditions that will reduce the risk of environmental contamination by R. equi and P. equorum eggs.     

Impacts of fluctuating temperature on the development and infectivity of entomopathogenic nematode Steinernema carpocapsae A10

Three different laboratory conditions were used to examine the impacts of fluctuating temperature on the development and infectivity of entomopathogenic nematode (EPN) Steinernema carpocaposae A10. Set I experiments focused on the impact of cold stress early in the development cycle. In these studies Galleria mellonella hosts were infected and incubated for 2 days at the control temperature of 23 degrees C and then subjected to lower temperatures of -10, 4, 10 or 14 degrees C, respectively, from days 3 to 36 post-infection (PI). Dissections of infected cadavers indicated arrested development at the adult stage at all lower temperatures tested. Set II experiments examined the impacts of cold stress early in the development followed by a return to 23 degrees C. Hosts were infected and incubated as in Set I and subjected to the same temperatures as above for 7 days, followed by incubation at 23 degrees C until 23 days PI. A limited number of EPN populations were able to complete development at 10 and 14 degrees C though emergent population numbers were significantly lower than those of control infections incubated continuously at 23 degrees C. In Set III experiments, infected hosts were subjected to cold stress later during development starting at day 4 post-infection followed by incubation at the control temperature. Population survival past first and second stage juveniles was reduced by at least 95% or more at the lower temperatures compared with controls. Emergent populations from the Set III cold-stressed hosts were not infectious. These studies may provide insights as to how EPN survive seasonal temperature fluctuations under natural environmental conditions.     

Persistence of Mycobacterium avium subsp. paratuberculosis and other zoonotic pathogens during simulated composting, manure packing, and liquid storage of dairy manure

Livestock manures contain numerous microorganisms which can infect humans and/or animals, such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., and Mycobacterium avium subsp. paratuberculosis (Mycobacterium paratuberculosis). The effects of commonly used manure treatments on the persistence of these pathogens have rarely been compared. The objective of this study was to compare the persistence of artificially inoculated M. paratuberculosis, as well as other naturally occurring pathogens, during the treatment of dairy manure under conditions that simulate three commonly used manure management methods: thermophilic composting at 55 degrees C, manure packing at 25 degrees C (or low-temperature composting), and liquid lagoon storage. Straw and sawdust amendments used for composting and packing were also compared. Manure was obtained from a large Ohio free-stall dairy herd and was inoculated with M. paratuberculosis at 10(6) CFU/g in the final mixes. For compost and pack treatments, this manure was amended with sawdust or straw to provide an optimal moisture content (60%) for composting for 56 days. To simulate liquid storage, water was added to the manure (to simulate liquid flushing and storage) and the slurry was placed in triplicate covered 4-liter Erlenmeyer flasks, incubated under ambient conditions for 175 days. The treatments were sampled on days 0, 3, 7, 14, 28, and 56 for the detection of pathogens. The persistence of M. paratuberculosis was also assessed by a PCR hybridization assay. After 56 days of composting, from 45 to 60% of the carbon in the compost treatments was converted to CO2, while no significant change in carbon content was observed in the liquid slurry. Escherichia coli, Salmonella, and Listeria were all detected in the manure and all of the treatments on day 0. After 3 days of composting at 55 degrees C, none of these organisms were detectable. In liquid manure and pack treatments, some of these microorganisms were detectable up to 28 days. M. paratuberculosis was detected by standard culture only on day 0 in all the treatments, but was undetectable in any treatment at 3 and 7 days. On days 14, 28, and 56, M. paratuberculosis was detected in the liquid storage treatment but remained undetectable in the compost and pack treatments. However, M. paratuberculosis DNA was detectable through day 56 in all treatments and up to day 175 in liquid storage treatments. Taken together, the results indicate that high-temperature composting is more effective than pack storage or liquid storage of manure in reducing these pathogens in dairy manure. Therefore, thermophilic composting is recommended for treatment of manures destined for pathogen-sensitive environments such as those for vegetable production, residential gardening, or application to rapidly draining fields.     

Temperature effect on Fusarium oxysporum f.sp. melonis survival during horticultural waste composting

AIMS: The aim of this work was to study the effect of high temperatures generated during composting process, on the phytopathogen fungus Fusarium oxysporum f.sp. melonis. This investigation was achieved by both in vivo (semipilot-scale composting of horticultural wastes) and in vitro (lab-scale thermal treatments) assays. METHODS AND RESULTS: Vegetable residues infected with F. oxysporum f.sp. melonis were included in compost piles. Studies were conducted in several compost windrows subjected to different treatments. Results showed an effective suppression of persistence and infective capacity, as this process caused complete fungal elimination after 2-3 days of composting. In order to confirm the effect of high temperature during this process, in vitro experiments were carried out. Temperature values of 45, 55 and 65 degrees C were tested. All three treatments caused the elimination of fungal persistence. Treatment at 65 degrees C was especially effective, whereas 45 degrees C eliminated fungal persistence only after 10 days. CONCLUSIONS: The composting process is an excellent alternative for the management of plant wastes after harvesting, as this procedure is able to suppress infective capacity of several harmful phytopathogens such as F. oxysporum f.sp. melonis. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium oxysporum f.sp. melonis is a plant pathogen fungus specially important in the province of Almería (south-east Spain), where intensive greenhouse horticulture is very extended. High temperatures reached during composting of horticultural plant wastes ensure the elimination of phytopathogen microorganisms such as F. oxysporum f.sp. melonis from vegetable material, providing an adequate hygienic quality in composts obtained.     

Effects of hyperthermia on DNA interstrand crosslinking after treatment with BCNU in 9L rat brain tumor cells

The effects of hyperthermia (42 degrees C) on 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU)-mediated DNA interstrand crosslink formation were investigated in 9L rat brain tumor cells using the technique of alkaline elution. When cells were treated with 60 microM BCNU for 1 hr at 37 degrees C and incubated for 6 hr in drug-free medium at 42 degrees C, there was a 50% increase in crosslinking; and when cells were treated at 42 degrees C and incubated at 37 degrees C, there was a 45% increase in crosslinking compared with the results for cells treated and incubated at 37 degrees C. When cells were treated and incubated at 42 degrees C, there was a 129% increase in DNA crosslinking. The same relative order of results was found for cell survival. These results suggest that hyperthermia can increase DNA interstrand crosslink formation and the consequent cell death through two independent mechanisms: an increase in the amount of initial alkylation because of the increased rate of hydrolysis of BCNU at higher temperatures, and the effect of heat on DNA structure that leads to an increase in the number of crosslinks formed.     

Continuous thermophilic composting (CTC) for rapid biodegradation and maturation of organic municipal solid waste

Fewer and fewer municipal solid wastes are treated by composting in China because of the disadvantages of enormous investment, long processing cycle and unstable products in a conventional composting treatment. In this study, a continuous thermophilic composting (CTC) method, only a thermophilic phase within the process, has been applied to four bench-scale composting runs, and further compared with a conventional composting run by assessing the indexes of pH, total organic carbon (TOC), total Kjeldahl nitrogen (TKN), C/N ratio, germination index (GI), specific oxygen uptake rate (SOUR), dissolved organic carbon (DOC) and dehydrogenase activity. After composting for 14 days, 16 days, 18 days and 19 days in the four CTC runs, respectively, mature compost products were obtained, with quality similar to or better than which had been stabilized for 28 days in run A. The products from the CTC runs also showed favorable stability in room temperature environment after the short-term composting at high temperature. The study suggested CTC as a novel method for rapid degradation and maturation of organic municipal solid wastes.     

Thermodynamics of reversible monomer-dimer association of tubulin

The equilibrium between the rat brain tubulin alpha beta dimer and the dissociated alpha and beta monomers has been studied by analytical ultracentrifugation with use of a new method employing short solution columns, allowing rapid equilibration and hence short runs, minimizing tubulin decay. Simultaneous analysis of the equilibrium concentration distributions of three different initial concentrations of tubulin provides clear evidence of a single equilibrium characterized by an association constant, Ka, of 4.9 X 10(6) M-1 (Kd = 2 X 10(-7) M) at 5 degrees, corresponding to a standard free energy change on association delta G degrees = -8.5 kcal mol-1. Colchicine and GDP both stabilize the dimer against dissociation, increasing the Ka values (at 4.5 degrees C) to 20 X 10(6) and 16 X 10(6) M-1, respectively. Temperature dependence of association was examined with multiple three-concentration runs at temperatures from 2 to 30 degrees C. The van't Hoff plot was linear, yielding positive values for the enthalpy and entropy changes on association, delta S degrees = 38.1 +/- 2.4 cal deg-1 mol-1 and delta H degrees = 2.1 +/- 0.7 kcal mol-1, and a small or zero value for the heat capacity change on association, delta C p degrees. The entropically driven association of tubulin monomers is discussed in terms of the suggested importance of hydrophobic interactions to the stability of the monomer association and is compared to the thermodynamics of dimer polymerization.