Studies on the growth metabolism of Bacillus thuringiensis and its vegetative insecticidal protein engineered strains by microcalorimetry

The metabolic power-times curves of Bacillus thuringiensis and its vegetative insecticidal protein engineered strains were determined at 30 degrees C by using a thermal activity monitor air Isothermal Microcalorimeter, ampoule method. From the power-times curves, the maximum power (Pmax) in the log phase, the growth rate constant (k), the generation times (tG), the time of the maximum power (tmax), the heat effects (Qlog) for log phase, and the total heat effect in 45 h (Qtotal) of B. thuringiensis strains can be obtained. The results indicate that their power-times curves are different. The relationship between their metabolic power-times curves and character of bacteria metabolism, and thermokinetics and gene expression were analyzed and discussed. The character of the bacteria power-times curves reflected the physiologic character of gene expression. The microcalorimetric method proved to be a reliable and sensitive tool for the assessment of the growth metabolism, the heat output in bacteria and its engineered strains. The determination of the thermokinetic character is beneficial to the control of fermentation.     

A thermoluminescence study of Photosystem II back electron transfer reactions in rice leaves – effects of salt stress

The recombination reactions of Photosystem II have been investigated in vivo in rice leaves by using the thermoluminescence (TL) emission technique. Excitation of dark-adapted leaf segments at 0 °C with different number of single turn-over flashes induced the appearance of complex TL glow curves. The mathematical analysis of these curves showed the existence of four TL components: B1-band (temperature maximum, t_max, at 24 °C, originating from S₃Q_B⁻ recombination), B2-band (t_max at 35 °C, from S₂Q_B⁻), AG-band (t_max at 46 °C) and C-band (t_max at 55 °C, from Tyr_D⁺Q_A⁻). Their contributions to the total TL signal were different depending on the number of flashes given. AG-band seems to reflect a special electron transfer from some unknown stroma donor to PS II. Q-band (t_max at 19 °C), originating from S₂Q_A⁻ recombination, was recorded after flashing samples incubated in the presence of DCMU. The recombination halftimes (t_1/2) at 20 °C of S₂Q_A⁻, S₃Q_B⁻, S₂Q_B⁻ and Tyr_D⁺Q_A⁻ were, respectively, 0.8 s, 48 s, 74 s and about 1 h. A sharp AG-band (t_max at 50 °C and t_1/2 of 210 s) could be also observed after illumination of leaves with far-red light and after a dark incubation period of whole plants. Incubation of leaf segments with 0.5 M NaCl abolished the inductions of AG-band by darkness and far-red illumination, significantly decreased Q-band intensity, whereas induced a strong increase in C-band intensity. The possible inhibition of S₂/S₃ formation and quinone oxidation by saline stress are discussed.     

Effect of Sm3+ ion on growth of Bacillus thuringiensis by microcalorimetry

By using an LKB-2277 Bioactivity Monitor, cycle-flow method, the thermogenic curves of aerobic growth for Bacillus thuringiensis cry II strain at 28°C have been obtained. The metabolic thermogenic curves of B. thuringiensis cry II contained two distinct patterns: the first reflects the changes during the bacterial growth phase and the second corresponds to the sporulation phase. From these thermogenic curves in the absence and presence of Sm³⁺ ions, the thermokinetic parameters such as the growth rate constants k, the interval time τ_I, the maximum power P _{max 1} and heat-output Q _log for log phase, the maximum power P _{max 2} and heat-output Q _stat for stationary phase, the heat-output Q _spor for sporulation phase and total heat effects Q _T are calculated. Sm³⁺ ion has promoting action on the growth of B. thuringiensis cry II in its lower concentration range; on the other hand, this ion has inhibitory action on the sporulation of B. thuringiensis in its higher concentration range. We also found that the effects of Sm³⁺ ion on B. thuringiensis during the sporulation phase were far greater than that during the bacterial phase. It is concluded that the application of B. thruringiensis of controlling insecticides is not affected by the presence of the rare-earth elements in the environmental ecosystem.     

Sorption of humic acids by bacteria

Capacity for sorption of humic acid (HA) from water solutions was shown for 38 bacterial strains. Isotherms of HA sorption were determined for the cells of 10 strains. The bonding strength between the cells and HA (k) and the terminal adsorption (Q _max) determined from the Langmuir equation for gram-positive and gram-negative bacteria were reliably different. Gram-positive bacteria sorbed greater amounts of HA than gram-negative ones (Q _max = 23 ± 10 and 5.6 ± 1.2 mg/m², respectively). The bonding strength between HA and the cells was higher in gram-negative bacteria than in gram-positive: k = 9 ± 5 and 3.3 ± 1.1 mL/mg, respectively.     

Thermoregulatory changes in the cold induced by physical training in humans

It has previously been demonstrated that metabolic heat production (M˙) during cold exposure at rest was related to maximal oxygen uptake (O_2max). Consequently, an increase in O_2max could allow an increase M˙ in the cold. The aim of the present study was therefore to test this hypothesis. Eight male volunteers undertook interval training (periods of 25% V˙O_2max of 30-s duration and 110% V˙O_2max of 60-s duration until exhaustion, five times a week over 8 weeks) to increase V˙O_2max. Both before and after this physical training, they were subjected to a 10^∘, 5^∘ and 1^∘C 2-h cold air test in a climatic chamber. During the cold exposure, rectal temperature (T _re), tympanic temperature (T _ty), mean skin temperature () and M˙ were measured as well as the time to onset of shivering (t) and body temperatures () at t. The results showed that physical training involved an increase in O_2max (14%–15%, P < 0.05). During the cold exposure, T _re was higher after training both at 10^∘,5^∘ and 1^∘C (P < 0.05) whereas were not significantly changed. However, an increase in the sensitivity of the thermoregulatory system was attested by a decreased t at higher These slight physiological changes found after training were not related to the increases in V˙O_2max. In conclusion, this study demonstrated that interval training induced slight thermoregulatory changes unrelated to changes in V˙O_2max and it suggested that M˙ during cold exposure could be related mainly to the level of V˙O_2max observed before training, since increases in V˙O_2max did not modify M˙. Accepted: 8 April 1998     

Shape of leaf photosynthetic electron transport versus temperature response curve is not constant along canopy light gradients in temperate deciduous trees

Responses of foliar light-saturated net assimilation rate (A_max), capacity for photosynthetic electron transport (J_max) and mitochondrial respiration rate (R_d) to long-term canopy light and temperature environment were investigated in a temperate deciduous canopy composed of Populus tremula L. in the upper (17–28 m) and of Tilia cordata Mill. in the lower canopy layer (4–17 m). Climatic measurements indicated that seasonal average daily maximum air temperature (T_max) was 5·5 °C (range 0·7–10·5 °C) higher in the top than in the bottom of the canopy, and strong positive correlations were observed between T_max and seasonal average integrated quantum flux density (Q_int), as well as between seasonal average daily mean temperature and Q_int. Because of changes in leaf dry mass and nitrogen per unit area, A_max, J_max, and R_d scaled positively with Q_int in both species at a common leaf temperature (T). According to J_max versus T response curves and dark chlorophyll fluorescence transients, photosynthetic electron transport was less heat resistant in P. tremula with optimum temperature of J_max, T_opt, of 33·5 ± 0·6 °C than in T. cordata with T_opt of 40·7 ± 0·6 °C. This difference was suggested to manifest evolutionary adaptation of photosynthetic electron transport to cooler environments in P. tremula, the range of which extends farther north than that in T. cordata. Possibly because of acclimation to long-term canopy temperature environment, T_opt was positively related to Q_int in P. tremula, foliage of which was also exposed to higher irradiances and temperatures, but not in T. cordata, in the canopy of which quantum flux densities and temperatures were lower, and gradients in the environmental factors less pronounced. Parallel to changes in T_opt, the activation energy for photosynthetic electron transport decreased with increasing Q_int in P. tremula, indicating that J_max of leaves acclimated to colder environment was more responsive to T in lower temperatures than that of high T acclimated leaves. Similar alterations in the activation energy for mitochondrial respiration rate were also observed, indicating that acclimation to temperature of mitochondrial and chloroplastic electron transport proceeds in a co-ordinated manner, and possibly involves long-term changes in membrane fluidity properties. We conclude that, because of correlations between temperature and light, the shapes of J_max versus T, and R_d versus T response curves vary within tree canopies, and this needs to be taken account in modelling whole canopy photosynthesis.     

A model of photosystem II for the analysis of fast fluorescence rise in plant leaves

The polyphasic patterns of fluorescence induction rise in pea leaves in vivo and after the treatment with ionophores have been studied using a Plant Efficiency Analyzer. To analyze in detail photosystem II (PS II) electron transfer processes, an extended PS II model was applied, which included the sums of exponential functions to specify explicitly the light-driven formation of the transmembrane electric potential (ΔΨ(t)) as well as pH in the lumen (pH_L(t)) and stroma (pH_S(t)). PS II model parameters and numerical coefficients in ΔΨ(t), pH_L(t), and pH_S(t) were evaluated to fit fluorescence induction data for different experimental conditions: leaf in vivo or after ionophore treatment at low or high light intensity. The model imitated changes in the pattern of fluorescence induction rise due to the elimination of transmembrane potential in the presence of ionophores, when ΔΨ = 0 and pH_L(t), pH_S(t) changed to small extent relative to control values in vivo, with maximum ΔΨ(t) ∼ 90 mV and ΔΨ(t) ∼ 40 mV for the stationary state at ΔpH ≅ 1.8. As the light intensity was increased from 300 to 1200 μmol m⁻² s⁻¹, the heat dissipation rate constants increased threefold for nonradiative recombination of P680⁺Phe⁻ and by ∼30% for P680⁺Q_A⁻. The parameters ΔΨ, pH_S and pH_L were analyzed as factors of PS II redox state populations and fluorescence yield. The kinetic mechanism of fluorescence quenching is discussed, which is related with light-induced lumen acidification, when ⁺Q_A⁻ and P680⁺ recombination probability increases to regulate the Q_A reduction.     

Maximum growth temperature ranges of Aeromonas Spp. isolated from clinical or environmental sources

Only a limited number of phenotypic tests are available for the differentiation of all 13 known hybridization groups (HG) of Aeromonas spp. These organisms have a wide spectrum of warm-blooded and cold-blooded hosts. In the present study, the maximum growth temperatures (t_max) of the most common HGs of Aeromonas spp. originating from human fecal samples, food, water, and healthy and diseased fish were determined with a plate-type continuous temperature-gradient incubator. We observed that determination of the t_max can be applied for differentiation of HG 1 from HG 2 and 3 (phenospecies A. hydrophila); HG 6 from HG 4, 5A, and 5B (phenospecies A. caviae); HG 7 from HG 8/10 (phenospecies A. sobria); and HG 11 from HG 8/10 (phenospecies A. veronii). HG 1, 4, 8/10, and 13 strains occurring also in human clinical samples had a high t_max, about 40°C or higher. Hybridization group 2, 3, 5A, and 5B strains, which in most cases originated from water or food, had t_max values in the range of about 36–39°C, while HG 6, 7, and 11 had t_max values in the range of about 33–37°C. Fish pathogenic strains of A. salmonicida subsp. salmonicida and subsp. achromogenes had the lowest t_max values from about 30 to 35°C. Correspondence to: M.-L. Hdnninen     

Antifungal activity of Bacillus thuringiensis strains and their efficacy against the cotton leaf worm Spodoptera littoralis

Bacillus thuringiensis strains that belong to B. thuringiensis, B. kurstaki and soil-isolated B.t. were assessed in the following phytopathogenic: Rhizoctonia solani, that had their mycelial growth decreased after incubation in the presence of the bacterial strains. The bacteria have also pathogenic effect against the insect pest Spodoptera littoralis. The isolate B.t. D-1 and the B.t. kurstaki HD-203 were found to be inhibiting R. solani, the strain B. kurstaki HD-203 displayed the highest percentage of inhibition (64%) and B.t. D-1 showed 49% of inhibition. Antagonistic activity was maintained up to pH 8.5, and the antifungal activity was stable to heat at 70?°C for 1?h. Minimal inhibitory concentrations were 152 and 131?μl/ ml for B.t. D-1 and B. kurstaki HD-203, respectively. The two strains also have high efficacy against S. littoralis larvae, B.t. D-1 gave 70% and the B. kurstaki HD-203 strain gave 80% mortality after seven days of treatment.     

Simplistic determination of operation parameters for simulated moving bed (SMB) chromatography for the separation of ketoprofen enantiomer

Since it is troublesome to estimate adequate flow rates in four sections of SMB chromatography, a systematic determination of the flow rates has been suggested by using ketoprofen as a model chiral enantiomer.S-ketoprofen, less retained species, was separated from raffinate stream and the variation in its purity was dependent on the changes of the flow rate of section 4 (Q ₄), the raffinate flow rate (Q _raf), and the feed flow rate (Q _feed) under a fixed switching timet ^*. When one parameter was changed at the given experimental condition, purities of product were changed and these phenomena has be well explained by the triangle theory.     

Evaluation of Bacillus thuringiensis bioinsecticidal protein effects on soil microorganisms

The effect of B. thuringiensis and its crystal protein on plant growth and on functional groups of microorganisms is not well understood. Soybean (Glycine max) var. Br 322 was grown in non-sterile soil infested with three B. thuringiensis (Bt) inocula: insecticidal crystal protein producer (Cry+), a mutant non-producer (Cry–), or insecticidal crystal protein (ICP), at a rate of 10⁷ cells g^–1 dry soil or 1.25 mg of protein g^–1 dry soil. Non-inoculated plants were maintained as control. Measurements were carried out on soil samples before sowing (time zero) and after sowing and inoculation (5, 15, 25, 35 and 45 d) on samples of rhizosphere soil. The effect of spore and crystal protein produced by B. thuringiensis on the populations of functional groups of microorganisms (bacteria including actinomycetes and fungi) involved in the biogeochemical cycling of carbon (cellulolytic, amylolytic and proteolytic), phosphorus (arbuscular mycorrhizal fungi), and nitrogen (number of nodules and proteolytic) were evaluated. Population sizes of culturable heterotrophic bacteria and saprophytic fungi were also evaluated. No difference was found in heterotrophic bacterial populations inoculated with B. thuringiensis. Difference was observed in functional groups of C-cycling microorganisms. Nodule formation and plant growth were increased by Cry+ strain and ICP when compared with uninoculated plants. Crystal protein did not show any effect on arbuscular mycorrhiza (AM) colonization. However, a deleterious effect was observed with Cry+ and Cry– strains that inhibited colonization of AM fungi when compared with uninoculated plants.     

Age at maturity of some fish species distributed in Turkish marine waters (Actinopterygii and Elasmobranchii)

We collected data on the age at maturity (t_m) and maximum reported age (t_max) for 153 stocks of marine fishes in Turkey, belonging to 59 species, 24 families and 2 classes (Actinopterygii and Elasmobranchii). Among Actinopterygii t_m had an average of 1.8 years (1 to 4 years) while among Elasmobranchii it had an average of 11.9 years (2 to 11.9 years). Overall, t_max ranged between two years (for Sarda sarda) and 34 years (for Squalus acanthias). Mean t_max was found to be 6.24 years for Actinopterygii and 10.11 years for Elasmobranchii. t_m showed a positive linear correlation with t_max for both Actinopterygii and Elasmobranchii. Mean t_m?t_max did not differ significantly with sex within the Actinopterygii and Elasmobranchii. The ratio t_m?t_max was found to be significantly lower for Actinopterygii than for Elasmobranchii.     

Effects of water stress on photosynthetic activity,dry mass partitioning and some associated metabolic changes in four provenances of neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis> A. Juss)

Chlorophyll (Chl) content, dry mass, relative water content (RWC), leaf mass per area (LMA), proline (Pro) content, malondialdehyde (MDA) content, superoxide dismutase (SOD) and peroxidase (POD) activity, P _N-PAR response curves and gas exchange were studied to determine the effects of water stress on photosynthetic activity, dry mass partitioning and metabolic changes in four provenances of neem (Azadirachta indica A. Juss). The results indicated that provenance differences existed in the adaptation response to water stress that included changes to growth strategies coupled with ecophysiological and metabolic adjustments. As water stress increased, stomatal conductance (g _s), net photosynthetic rate (P _N), transpiration rate (E), and leaf RWC decreased while LMA increased in all provenances. Dry mass was reduced in droughted plants and the percentage increased in dry mass allocated to roots, and enzyme activities of SOD and POD were highest in neem originating from Kalyani (KA) provenance and lowest in neem originating from New Dehli (ND) provenance. In contrast, water stress increased MDA content least in KA and most in ND. Furthermore, neem originating from ND also had the greatest decrease in Chl a/b ratio while the ratio was least affected in neem originating from KA. These findings suggest neem originating from KA may have more drought resistance than neem originating from ND. The data from P _N-PAR response curves are less clear. While these curves showed that drought stress increased compensation irradiance (I _c) and dark respiration (R _D) and decreased saturation irradiance (I _s) and maximum net photosynthetic rate (P _max), the extent of decline in P _max was provenance dependent. P _max under non-waterlimiting conditions was higher in neem originating from Jodhpur (MA) (about 14 μmol m⁻² s⁻¹) than in the other three provenances (all about 10 μmol m⁻² s⁻¹), but mild water stress had minimal effect on P _max of these three provenances whereas P _max of MA provenance declined to 10 μmol m⁻² s⁻¹, i.e. a similar value. However, under severe water stress P _max of MA and KA provenances had declined to 40% of non-stressed values (about 6 and 4 μmol m⁻² s⁻¹, respectively) whereas the decline in P _max of neem originating from Kulapachta (KU) and ND provenances was about 50% of nonstressed values (about 5 μmol m⁻² s⁻¹). These data suggest the P _N responses of KU and ND provenances are most tolerant, and KA and MA least tolerant to increasing water stress, but also suggest MA provenance could be the most desired under both non-water-limiting and water-limiting conditions due to highest P _max in all conditions.     

Performance assessment of malolactic fermenting bacteria Oenococcus oeni and Lactobacillus brevis in continuous culture

The growth performance of malolactic fermenting bacteria Oenococcus oeni NCIMB 11648 and Lactobacillus brevis X₂ was assessed in continuous culture. O. oeni grew at a dilution rate range of 0.007 to 0.052 h⁻¹ in a mixture of 5:6 (g l⁻¹) of glucose/fructose at an optimal pH of 4.5, and L. brevis X₂ grew at 0.010 to 0.089 h⁻¹ in 10 g l⁻¹ glucose at an optimal pH of 5.5 in a simple and safe medium. The cell dry weight, substrate uptake and product formation were monitored, as well as growth kinetics, yield parameters and fermentation balances were also evaluated under pH control conditions. A comparison of growth characteristics of two strains was made, and this showed significantly different performance. O. oeni has lower maximum specific growth rate (μ_max=0.073 h⁻¹), lower maximum cell productivity (Q _x ^max=17.6 mg cell l⁻¹ h⁻¹), lower maximum biomass yield (Y _x/s ^max=7.93 g cell mol⁻¹ sugar) and higher maintenance coefficient (m _s=0.45 mmol⁻¹ sugar g⁻¹ cell h⁻¹) as compared with L. brevis X₂ (μ_max=0.110 h⁻¹; Q _x ^max=93.2 g⁻¹ cell mol⁻¹ glucose; Y _x/s ^max=22.3 g cell mol⁻¹ glucose; m _s=0.21 mmol⁻¹ glucose g⁻¹ cell h⁻¹). These data suggest a possible more productive strategy for their combined use in maturation of cider and wine.     

Daily rhythm of oxygen consumption and thermoregulatory responses in some European winter- or summer-acclimatized finches at different ambient temperatures

The oxygen consumption of European finches, the siskin (Carduelis spinus), the brambling (Fringilla montifringilla), the bullfinch (Pyrhulla pyrhulla), the greenfinch (Carduelis chloris) and the hawfinch (Coccothraustes coccothraustes), was recorded continuously while ambient temperature was decreased stepwise from +30 down to-75°C. The oxygen consumption, body temperature (telemetrically), and shivering (integrated pectoral electromyography) of greenfinches were measured simultaneously at ambient temperatures between +30 and-75°C. Maximum heat production, cold limit, lower critical temperature, basal metabolic rate and thermal conductance (of the greenfinch) were determined. The diurnal variation of oxygen consumption of siskins and greenfinches was recorded at thermoneutrality and below the thermoneutral zone in winter- and summer-acclimatized birds. The diurnal variation of body temperature and thermal conductance of greenfinches were also determined. The diurnal variation of heat production was not seasonal or temperature dependent in the siskin and in the greenfinch. Nocturnal reduction of oxygen consumption saved 15–33% energy in the siskin and greenfinch. Body temperature of the greenfinch was lowered by 2.5–3.4°C. The nocturnal reduction of thermal conductance in the greenfinch was 39–48%. The basal metabolic rate was lowest in the largest bird (hawfinch) and highest in the smallest bird (siskin). The values were in the expected range. The heat production capacity of finches in winter was 4.7 times basal metabolic rate in the siskin, 4.2 times in the brambling, 3.5 times in the greenfinch and 2.9 times in the bullfinch and hawfinch. The heat production capacity of the siskin and greenfinch was not significantly lower in summer. The cold limit temperatures (°C) in winter were-61.2 in the siskin,-41.3 in the greenfinch,-37.0 in the bullfinch,-35.7 in the brambling and-28.9 in the hawfinch. The cold limit was 14.3°C higher in summer than in winter in the siskin and 8.7°C in the greenfinch. Thermal insulation of the greenfinch was significantly better in winter than in summer. The shivering of the greenfinch increased linearly when ambient temperature was decreased down to-40°C. Maintenance of shivering was coincident with season. In severe cold integrated pectoral electromyography did not correlate with oxygen consumption as expected. The possible existence of non-shivering thermogenesis in birds is discussed. It is concluded that the acclimatization of European finches is primarily metabolic and only secondly affected by insulation.Abbreviations AAT avian adipose tissue - bm body mass - BMR basal metabolic rate - C _t thermal conductance - EMG electromyogram - HP heat production - HP _max maximum heat production - MR metabolic rate - NST non-shivering thermogenesis - RMR resting metabolic rate - RQ respiratory quotient - T _a ambient temperature - T _b body temperature - T _c colonic temperature - T _1c lower critical temperature - TNZ thermoneutral zone - T _st shivering threshold temperature - V oxygen consumption     

Detection of enterotoxin genes in mosquito-larvicidal Bacillus species

The presence of the B component of hemolysin BL (hblA), enterotoxin BceT (bceT), and enterotoxin S (entS) genes in mosquito-larvicidal Bacillus sp., including 25 B. sphaericus and 4 B. thuringiensis subsp. israelensis strains, has been analyzed by multiplex PCR in this study. The results showed that all four B. thuringiensis strains contain the hblA gene and the sequences of bceT and entS genes. However, none of the enterotoxin gene sequences were detected in the B. sphaericus strains. The enterotoxin production in all strains has also been analyzed by using two commercial immunoassay kits (TECRA and RPLA), and it has been proved that all the B. thuringiensis strains and one B. cereus strain can produce enterotoxins during growth. No enterotoxin activity could be detected in B. sphaericus strains. Received: 20 November 2001 / Accepted: 31 December 2001     

Antibacterial Activities of Novel Diselenide-bridged <Emphasis Type="Italic">bis</Emphasis>(Porphyrin)s on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> Investigated by Microcalorimetry

The actions of two novel diselenide-bridged bis(porphyrin)s (1 and 2) on Staphylococcus aureus growth was investigated by microcalorimetry at 37.00°C, compared with that of Na₂SeO₃. Differences in their capacities to inhibit the growth metabolism of S. aureus were observed. By analyzing the power–time curves, crucial parameters such as the rate constant of bacterial growth (k), inhibitory rate (I), and generation time (t _G) were determined. The growth rate constant (k) of S. aureus (in the log phase) in the presence of the drugs decreased with increasing concentrations of the drugs regularly. The relationship of k and c is nearly linear for diselenide-bridged bis(porphyrin) 2. The sequence of the antibacterial activities of these selenium compounds tested was 2 > 1 > Na₂SeO₃.     

Evaporative water loss: thermoregulatory requirements and measurements in the deer mouse and white rabbit

Summary Using a physical model of the capacity for non-evaporative heat loss and measurements of metabolic heat production, I evaluated the evaporative requirements for thermoregulation in the deer mouse,Peromyscus maniculatus, and the white rabbit,Oryctolagus cuniculus. The physical limit to non-evaporative heat loss was calculated from the heat transfer properties of the two animals and expressed as a maximum thermal conductance (C _max). Two physiologically-based thermal conductances were derived from evaporative water loss, respiratory gas exchange and core temperature measurements made between 8 and 34°C on the deer mouse, and taken from published data for the white rabbit. The thermal conductance for non-evaporative heat loss (C) was calculated from net heat production, whereasC _m represented the thermal conductance required to dissipate metabolic heat production. Evaporation is required when metabolic heat production exceeds the capacity for non-evaporative heat loss (as shown byC _m>C _max). However, evaporation increased in both animals although additional capacity to lose heat remained (i.e.,C<C _max). Evaporation increased withC above 30°C for the mouse and at each 5°C measurement interval from 15 to 30°C for the rabbit. Thus, evaporation was greater than that required for thermoregulation for both animals as determined from a physical model of heat loss because both evaporation andC increased together to regulate heat loss.Symbols: see list on title page of preceeding paper     

The Matched Pairs Problem with Exponential Variates

Two classes of tests for the hypothesis of bivariate symmetry are studied. For paired exponential survival times (t_1j, t_2j), the classes of tests are those based on t_1j-t_2j and those based on log t_1j–log t_2j. For each class the sign, signed ranks, t and likelihood ratio tests are compared via Pitman's criterion of asymptotic relative efficiency (ARE). For tests based on t_1j — t_2j, it is found that: (1) the efficacy of the paired t depends on the coefficient of variation (CV) of the pair means, (2) the signed rank test has the same ARE to the sign test as for the usual location problem. For tests based on log t_1j — log t_2j, the ARE comparisons reduce to the well-known results for the one-sample location problem for samples from a logistic density. Hence, the signed rank test is asymptotically efficient. Furthermore, analyses based on log t_1j — log t_2j are not complicated by the underlying pairing mechanism.