Thermal conductance of guanaco (Lama guanicoe) pelage

1. Guarded hot plate technique was used to measure thermal conductance of winter, summer and sheared pelts of Lama guanicoe.

2. Mean thermal conductance of winter and summer pelage was 1.64 W/m² °C and 1.79 W/m² °C, respectively.

3. Mean heat loss in natural pelage, in free convection conditions was 1.74 W/m² °C and 2.3 W/m² °C in sheared pelts.

4. From our results, there is seasonal moult in guanacos.     

A simple in-vitro 'wet-plate' method for mass production of Phytophthora nicotianae zoospores and factors influencing zoospore production

A simple in-vitro ‘wet-plate’ method for mass-producing Phytophthora nicotianae zoospores at ≥ 1.0 × 10⁶ zoospores/ml is described. Temperature critically affected zoospore production; 22 °C was optimum, while 36 °C was completely inhibitory. Zoospores being the most important propagule of P. nicotianae, temperature of recycled irrigation water may be manipulated to reduce diseases in irrigated nursery crops.     

Critical thermal maximum and body water loss in first instar larvae of three Cetoniidae species (Coleoptera)

Critical thermal maximum (CT_max) and body water losses were measured in first instar larvae of Gnorimus nobilis, Osmoderma eremita (Trichiinae) and Cetonischema aeruginosa (Cetoniinae) when air temperature was increased gradually (0.5 °C/min) from 20 °C to the critical thermal maximum (CT_max), in dry air (near 0% R.H.).

The CT_max was significantly lower in O. eremita (45.6±0.7 °C) than in G. nobilis (48.5±0.6) and C. aeruginosa (51.4±0.9 °C).

An increase of 10 °C (30–40 °C) induced a 2-fold increase of the water loss in C. aeruginosa and O. eremita (Q₁₀=2.10±0.12 and 2.13±0.20, respectively). In the range from 40 to 45 °C to CT_max a strong increase of the water loss was observed in O. eremita and C. aeruginosa, respectively. Body water losses were significantly lower in C. aeruginosa than in O. eremita and G. nobilis over the range 20 °C—CT_max; no significant difference occurred between G. nobilis and O. eremita.     

The effect of constant and changing temperatures on the thermal resistance of Lymnaea peregra (Müller)

The effect of constant and changing temperatures on both high and low lethal temperatures of Lymnaea peregra and the time course of acclimation were investigated. Snails acclimated to 6.5°, 11.5° and 16.5°C showed paradoxical ‘adaptation’ of their high letal temperature and reasonable ‘adaptation’ of the lowlethal temperature. This combination is thus observed for the first time in molluscs. Seasonal variation in the upper lethal temperature showed parallel changes and the snails became less resistant to heat during spring and summer. Acclimation to changing temperature conditions did not extend their temperature tolerance range. During the course of acclimation of 6.5°C snails to 16.5°C, about 70% of the acclimation was completed within the first 24 h and the rest was completed within 360 h at 16.5°C.     

Geographic variation in field body temperature of sceloporus lizards

1. Using data from the literature, I assessed how broad climatic patterns affected field body temperatures (T_b’s) of lizards in the genus Sceloporus.

2. Sceloporus at temperate latitudes had mean T_b’s of 35°C throughout their elevational range. This pattern is associated with “tropical” temperatures that extend into high north latitudes during the summer and the relatively low elevations occupied by the lizards.

3. At tropical latitudes, mean T_b declined from 35°C at low elevations to 31°C at high elevations. This pattern is associated with low seasonal variation in temperature at tropical latitudes and the relatively high elevations occupied by the lizards.     

Thermoregulatory use of heat increment of feeding in the tawny owl (Strix aluco)

The heat increment of feeding (HIF) was investigated in the tawny owl (Strix aluco) in central Norway (63°N, 10°E), close to the northern limit of its distribution. HIF was measured as the increase in heat production (measured as oxygen consumption) after force-feeding the owls with laboratory mice at thermoneutral conditions (20 °C) and during cold-exposure (5 °C and −5 °C). The basal metabolic rate of the owls (mean mass 419 g) was 4.39 kJ h⁻¹ and the lower critical temperature was approximately 16 °C. During cold conditions, HIF substituted for thermogenesis, and at an ambient temperature of −5 °C the substitution was complete. Calculations indicate that the substitution by HIF may save the owls as much as 60% of their daily thermoregulatory costs. This corresponds to about 10% of their total daily energy budget.     

The effects of constant and changing tempertures on the development of eggs of the freshwater snail Lymnaea auricularia (L.)

The rate of development of Lymnaea auricularia eggs was studied at various constant temperatures between 10° and 36°C. Development was accelerated as the temperature increased and at 36°C the eggs failed to develop. Spring eggs showed differences in their rate of development when compared with summer eggs when measured at similar tempertures.

Both spring and summer eggs were more than 90% fertile. Hatching success was high at temperatures between 10° and 30° (100%–82/9%); while at 34°C it was reduced to 60.6% for spring eggs. It was above 87% at temperatures between 10° and 34°C but it dropped to 62.3% at 36°C for summer eggs.

In one regularly changing temperature experiment a significant acceleration (P < .05) was found. In two others there was no significant difference beween predicted and observed egg durations. In one suddenly changing temperature regime (1 day at 20°, 1 day at 30° and so on) a huge retardation of development was found. In the other suddenly changing experiment (1 day at 15°, 1 day at 25°) no significant difference was found.

The exposure of eggs to extreme temperature (4°C, freezing and 4°C caused a retardation in the race of subsequent development of eggs at 25°C.     

Temperature-mediated characteristics of the dusky salamander (Desmognathus fuscus) of southern Appalachia

Thermal preference of the salamander Desmognathus fuscus was measured in a linear thermal gradient with floor temperatures ranging from 10 to 30°C. Salamanders were acclimated to 21±1°C and a 12 : 12 photoperiod with photophase centered at 1200 h for 8 weeks prior to being placed in the gradient. Substrate temperatures were measured under the salamanders’ stomachs from 1200 to 2400 h at 2 h intervals immediately after feeding and after seven days fasting. We found no selection for temperature in fasting or postprandial D. fuscus. We compared the rate at which D. fuscus cooled and heated with that of a control and found no significant difference. We determined the desiccation rate of D. fuscus at 16 and 26°C and found a significantly more rapid desiccation at 26°C.     

Effect of heat–moisture treatment on the structure and physicochemical properties of tuber and root starches

Starch from tubers potato (Solanum tuberosum), taro (Alocassia indica), new cocoyam (Xanthosoma sagitifolium), true yam (Dioscorea alata), and root cassava, (Manihot esculenta) crops was isolated and its morphology, composition and physicochemical properties were investigated before and after heat–moisture treatment (HMT) (100 °C, for 10 h at a moisture content of 30%). Native starch granules were round to oval to polygonal with smooth surfaces. The granule size (diameter) ranged from 3.0 to 110 μm.The total amylose content ranged from 22.4 to 29.3%, of which 10.1–15.5% was complexed by native lipid. The phosphorus content ranged from 0.01 to 0.1%. The X-ray pattern of potato and true yam was of the ‘B’-type. Whereas, that of new cocoyam and taro was of the ‘A’-type. Cassava exhibited a mixed ‘A+B’-type X-ray pattern. The relative crystallinity, swelling factor (SF), amylose leaching (AML), gelatinization temperature range and the enthalpy of gelatinization of the native starches ranged from 30 to 46, 22 to 54, 5 to 23%, 13 to 19 °C and 12 to 18 J/g, respectively. Susceptibility of native starches towards hydrolysis by 2.2N HCl and porcine pancreatic -amylase were 60–86% (after 12 days), and 4–62% (after 72 h), respectively. Retrogradation was most pronounced in the B-type starches. Granule morphology remained unchanged after HMT. The X-ray pattern of the B-type starches was altered (B→A+B) on HMT. However, that of the other starches remained unchanged. HMT decreased SF, AML, gelatinization enthalpy and susceptibility towards acid hydrolysis, but increased gelatinization temperatures and enzyme susceptibility. Extent of retrogradation and relative crystallinity decreased on HMT of true yam and potato starches, but remained unchanged in the other starches. The foregoing data showed that changes in physicochemical properties on HMT are influenced by the interplay of crystallite disruption, starch chain associations and disruption of double helices in the amorphous regions.     

Continuous-flow complete-mixing system for assessing the effects of environmental factors on colony-level coral metabolism

A small-scale chamber experimental system was designed to study the effects of temperature on colony-level coral metabolism. The system continuously supplies fresh seawater to the chamber, where it is mixed immediately and completely with the seawater already present. This continuous-flow complete-mixing system (CFCM system), in conjunction with theoretical equations, allows quantitative determination of chemical uptake and release rates by coral under controlled environmental conditions. We used the massive hermatypic coral Goniastrea aspera to examine variations in pH, total alkalinity, and total inorganic carbon for 16 days at 27 °C under controlled light intensities (300 and 0 µmol m^− 2 s^− 1). We confirmed the stability of the CFCM system with respect to coral photosynthetic and calcification fluxes. In addition, we obtained daily photosynthetic and calcification rates at different temperatures (27 °C, 29 °C, 31 °C, and 33 °C). When seawater temperature was raised from 31 °C to 33 °C, the gross primary production rate (P_gross) decreased 29.5%, and the calcification rate (G) decreased 85.7% within 2 days. The CFCM system allows quantitative evaluation of coral colony chemical release and uptake rates, and metabolism.     

Effect of nifedipine on core cooling in rats during tail cold water immersion

Male rats (450 g, n=11/group) were heated at an ambient temperature of 42°C until a rectal temperature of 42.8°C was attained. Rats, then received either saline (30°C)+tail ice water immersion (F+I) or saline (30°C)+tail ice water immersion+Nifedipine, a peripheral vasodilator, (F+I+N) to determine cooling rate effectiveness and survivability. The time to reach a rectal temperature of 42.8°C averaged 172 min in both groups resulting in similar heating rates (0.029°C/min). The cooling rates in group F+I and F+I+N were not significantly different from each other. We conclude that since Nifedipine did not improve cooling rates when combined with fluid+tail ice water immersion, its use as a cooling adjunct does not seem warranted.     

Morphology and germination characteristics of the cysts of Chattonella ovata (Raphidophyceae), a novel red tide flagellate in the Seto Inland Sea, Japan

To elucidate the mechanism of bloom outbreaks of Chattonella ovata (Raphidophyceae), we investigated the cysts of C. ovata and succeeded in finding them from the bottom sediments of Hiroshima Bay. The morphology of the cysts was mostly hemispherical in shape, with a diameter of ca. 30 μm and height of ca. 20 μm. The cysts were usually adhering to solid materials, such as diatom frustules, yellow-greenish in color and had several dark brown grains. The cyst wall was smooth and had no ornamentation. Because the morphological characteristic of the cysts was in general agreement with those of Chattonella antiqua and Chattonella marina, it was difficult to differentiate the cysts of these three species. Germination of the cysts of C. ovata was observed at temperatures from 17.5 to 30 °C, but not at 15 °C or below. The number of the germinated cysts increased with increasing temperature and the optimum temperature for germination was 30 °C. Although cysts of C. antiqua and C. marina germinated at temperatures from 15 to 30 °C, optimum temperature of germination was 22.5 °C. The lower limit and optimum temperatures for germination of C. ovata cysts was higher than for C. antiqua and C. marina. The role of cysts in the population dynamics of C. ovata is discussed.     

Extraction of defatted rice bran by subcritical water treatment

Defatted rice bran was treated with subcritical water in the temperature range of 180–280 °C for 5 min using 117 mL and 9 mL vessels to produce the extracts. The total sugar and protein contents and radical scavenging activity of the extracts were then estimated for both vessels. The total sugar concentration of ca. 0.3 g/L-extract was the highest for the extracts at 200 °C, and it significantly decreased at the higher temperatures. The protein concentration and radical scavenging activity were higher at the higher temperatures. Extraction was also done at 200 °C and 260 °C for various times using the small vessel. The total sugar concentration decreased with the increasing extraction time, while the protein concentration and radical scavenging activity only slightly depended on the extraction time. The extracts at 200 °C or lower temperatures using the large vessel possessed the emulsifying and emulsion-stabilizing activities. The HPLC analysis of the extract at 260 °C for 5 min using the small vessel indicated that it contained both hydrophilic and hydrophobic substances. The hydrophilic fraction of the extract mainly contained low-molecular-mass substances.     

Isolation and characterization of a metagenome-derived and cold-active lipase with high stereospecificity for (R)-ibuprofen esters

We report on the isolation and biochemical characterization of a novel, cold-active and metagenome-derived lipase with a high stereo-selectivity for pharmaceutically important substrates. The respective gene was isolated from a cosmid library derived from oil contaminated soil and designated lipCE. The deduced aa sequence indicates that the protein belongs to the lipase family l.3, with high similarity to Pseudomonas fluorescens lipases containing a C-terminal secretion signal for ABC dependent transport together with possible motifs for Ca²⁺-binding sites. The overexpressed protein revealed a molecular weight of 53.2 kDa and was purified by refolding from inclusion bodies after expression in Escherichia coli. The optimum temperature of LipCE was determined to be 30 °C. However, the enzyme still displayed 28% residual activity at 0 °C and 16% at −5 °C. Calcium ions strongly increased activity and thermal stability of the protein. Further detailed biochemical characterization of the recombinant enzyme showed an optimum pH of 7 and that it retained activity in the presence of a range of metal ions and solvents. A detailed analysis of the enzyme's substrate spectrum with more than 34 different substrates indicated that the enzyme was able to hydrolyze a wide variety of substrates including the conversion of long chain fatty acid substrates with maximum activity for pNP-caprate (C₁₀). Furthermore LipCE was able to hydrolyze stereo-selectively ibuprofen-pNP ester with a high preference for the (R) enantiomer of >91% ee and it demonstrated selectivity for esters of primary alcohols, whereas esters of secondary or tertiary alcohols were nearly not converted.     

Extraction of polyhydroxyalkanoate from Sinorhizobium meliloti cells using Microbispora sp. culture and its enzymes

Sinorhizobium meliloti produced 50% polyhydroxyalkanoate (PHA) in the biomass in the presence of sucrose as carbon substrate. Isolation of the intracellular PHA was achieved through a secondary fermentation involving a cell lytic actinomycetes species namely Microbispora sp. without further supplementation of nutrients to the S. meliloti fermented broth, at 30 °C, 150 rpm up to 72 h. Microbispora sp. cells that showed pelleted growth was removed by filtration and the released polymer contained in the filtrate was extracted by chloroform or an admixture of Triton X 100 (0.6%) a surfactant and ethylene diamine tetra acetic acid (EDTA) a chelating agent. Yield of PHA obtained was 49, 41 and 7% of biomass weight after 24, 48 and 72 h of lytic culture fermentation, respectively. Corresponding recovery of the polymer was 94, 82 and 15% of 90% purity. Alternatively Microbispora sp. lytic enzyme was obtained by its cultivation in nutrient broth with S. meliloti cells as substrate and the supernatant was used for the hydrolysis of the PHA containing biomass to release PHA. A620 lytic activity value for the broth was 200 at 72 h. The enzyme showed optimized activity at 50 °C, pH 7 and this was used to hydrolyze 5 g/l of thermally inactivated biomass of S. meliloti to recover 94% of total PHA present in the cells and the polymer produced was 92% pure. Decreased cell lytic activity in the presence of soluble protein added in the form of bovine serum albumin indicated that the hydrolytic activity may be due to proteases. The polymer was characterized by GC, NMR and DSC and was found to be polyhydroxybutyrate-co-hydroxyvalerate (97:3 mol%) with a melt temperature of 169 °C.     

Latent infection: a low temperature survival strategy in steinernematid nematodes

1. Entomopathogenic nematodes penetrate and kill Galleria mellonella within 48 h at optimal temperatures.

2. Low temperature induces infection latency, preventing host death until optimal conditions resume.

3. Infected Galleria survived 25 days at 5°C. On transfer to 25°C, 100% and 12.5% of Steinernema carpocapsae and Steinernema riobravis infected larvae died within 72 h.

4. Infective juvenile penetration decreased with decreasing temperature; declining from 49.7 and 49.3 nematodes/host at 25°C to 6.3 and 0.25 nematodes/host at 5°C for S. carpocapsae and S. riobravis, respectively.

5. Latent infection occurs, albeit infrequently, due to low host penetration at low temperature.

Determination of the thermodynamics of carbonic anhydrase acid-unfolding by titration calorimetry

The enthalpy of unfolding (Δ_uH) of carbonic anhydrase II was determined by titrating the protein with acid and measuring the heat using isothermal titration calorimetry (ITC) in the temperature range of 5 to 59 °C. By combining the ITC results with our previous findings by differential scanning calorimetry (DSC) in the temperature range of 39 to 72 °C, the Δ_uH dependence over a wide temperature range was obtained. The temperature dependence of the enthalpy displays significant curvature indicating that the heat capacity of unfolding (Δ_uC_p) is dependent on temperature. The T-derivative of Δ_uC_p was equal to 100 ± 30 J/(mol × K²), with the result that the Δ_uC_p is equal to 15.8 kJ/(mol × K) at 5 °C, 19.0 kJ/(mol × K) at 37 °C and 21.8 kJ/(mol × K) at 64 °C. The enthalpy of unfolding is zero at 17 °C. At lower temperatures, the Δ_uH becomes exothermic.

This method of determining protein unfolding thermodynamics using acid-ITC, significantly widens the accessible T-range, provides direct estimate of the thermodynamic parameters at physiological temperature, and gives further insight into the third T-derivative of the Gibbs free energy of unfolding.     

Effects of feeding supplemental palmitic acid (C16:0) on performance and milk fatty acid profile of lactating dairy cows under summer heat

The objective was to determine performance and milk fatty acid changes of high producing dairy cows in early lactation, under summer heat, by adding a supplemental rumen inert fat in the form of a saturated free fatty acid (856 g/kg C16:0/kg of total fatty acids) to the total mixed ration (TMR). Early lactation multiparous Holstein cows in two similar pens of 99 and 115 cows were used in a 2 × 2 Latin Square design experiment with 35 d periods during a period when daily high and low temperatures averaged 34.3 and 15.9 °C, the relative humidity averaged 51% and there were no rain events. The TMR was the same for both groups, consisting of approximately 435 g/kg forage and 565 g/kg concentrate, except that the vitamin/mineral premix had no added fat (control, C) or added fat (C16:0) at a level designed to deliver approximately 450 g/cow/d of supplemental fat if cows consumed 26.5 kg/d of dry matter (DM). The two TMR averaged 905 g/kg organic matter (OM), 318 g/kg neutral detergent fiber (aNDF), and 186 g/kg crude protein (CP). The ‘C’ TMR had 58 g/kg total fatty acids with an estimated net energy for lactation (NE_l) of 7.3 MJ/kg (DM), while the C16:0 TMR had 72 g/kg total fatty acids and 7.5 MJ/kg NE_l (DM). Whole tract digestibility of DM, OM, aNDF and CP tended (P<0.10) to increase, and that of fatty acids increased substantially (P<0.01), with C16:0 feeding, whereas, DM intake was not affected. Milk fat content decreased (P<0.01) with C16:0 feeding (37.5 versus 36.0 g/kg), whereas, true protein content tended (P=0.09) to increase. There was a tendency (P=0.07) for increased milk yield (36.69 versus 38.04 kg/d), while milk protein yield increased (P=0.03) with C16:0 supplementation (1.08 versus 1.13 kg/d). Milk fat yield was unaffected by treatment. Concentrations of short and medium chain milk fatty acids (C6:0–C15:0), decreased, or tended to decrease, with C16:0 addition (C13:0 and C15:0, P<0.10; all others, P≤0.05). The concentration of C16:0 increased (P<0.001) in milk triglycerides from cows fed C16:0 (27.10 versus 31.57 g/kg), the longer chain saturated fatty acids C17:0 and C18:0 decreased (P≤0.05) and other long chain unsaturated fatty acids were unaffected. Benefits of C16:0 feeding on cow productivity must be balanced against negative effects on the nutritive value of the milk (i.e., increased C16:0 in milk fatty acids) produced for human consumption. However, relatively low amounts of supplemental C16:0 (27.10 versus 31.57 g/kg in milk triglycerides for C and C16:0 supplemented cows, respectively) were actually secreted in milk, in spite of them being essentially fully digested in the digestive tract. Strategies to divide cows into production groups based on milk yield and/or milk fat proportions could further limit C16:0 secretion in milk. Supplemental dietary C16:0 may have positive effects on milk production that outweigh the negative health effects of the increased C16:0 content in the milk fat.     

Stress physiological responses to tourist pressure in a wild population of European pine marten

The tourist pressure in natural parks is a potential source of stress and may cause an increase in the adrenal activity of wild populations of European pine marten (Martes martes). Seventy-six faecal samples were collected during 15 months in a natural park of Northwest Spain. Analysis of faecal DNA was used for the specific identification using the PCR-RFLPs technique. Faecal steroid determinations were performed by EIA. Natural park was divided in three areas: free entry, restricted area, and integral reservation, and number of daily human visitors recorded. Faecal glucocorticoid metabolite levels (ng/g dry faeces) were significantly higher in spring (56.36 ± 19.62) and summer (31.27 ± 11.98) compared to autumn (15.33 ± 6.89) and winter (11.13 ± 3.30). These data are closely related to daily number of visitors (spring: 3204, summer: 1672, winter: 646, autumn: 551). Androgen, progestin and oestrogen levels were also significantly higher in spring (reproductive season) showing values of 43.62 ± 18.6, 154.31 ± 53.50 and 829.62 ± 456.1, respectively. Glucocorticoid levels were significantly lower in integral reservation (15.95 ± 3.56) compared to restricted (31.4 ± 16.30) and free entry areas (41.59 ± 12.73), respectively. Wild populations of European pine marten showed stress physiological response induced by the tourist pressure and this response is higher during reproductive season.     

Determination of the maximum and minimum lethal temperatures (LT50) for Loxosceles intermedia Mello-Leitão, 1934 and L. laeta (Nicolet, 1849) (Araneae, Sicariidae)

In this study, the maximum and minimum lethal temperatures (LT₅₀) of L. intermedia and L. laeta were determined in two treatments: gradual heating (25–50°C) and cooling (25°C to −5°C), and 1 h at a constant temperature. In gradual temperatures change, L. intermedia mortality started at 40°C and the LT₅₀ was 42°C; for L. laeta, mortality began at 35°C and the LT₅₀ was 40°C. At low temperatures, mortality was registered only at −5°C for both species. In the constant temperature L. intermedia showed a maximum LT₅₀ at 35°C and L. laeta at 32°C; the minimum LT for both species was −7°C.