Polyunsaturated dietary lipids lower the selected body temperature of a lizard

Summary Cold acclimation lowers the selected body temperature (T _b) in many ectothermic vertebrates. This change in behavioural thermoregulation is accompanied by an increase in the proportion of polyunsaturated fatty acids in tissues and cellular membranes. We investigated how diets containing different fatty acids, known to significantly alter the fatty acid composition of animal tissues and membranes, affect the selected T _b of the lizard Tiliqua rugosa. Lizards on a diet containing many polyunsaturated fatty acids (10% sunflower oil) showed a 3–5°C decrease in T _b, whereas T _b in animals on a diet containing mainly saturated fatty acids (10% sheep fat) did not change. Our study suggests that the composition of dietary lipids influences thermoregulation in ectothermic vertebrates and may thus play a role in the seasonal adjustment of their physiology.Abbreviations CST central standard time - T _a air temperature - T _b Body temperature     

THERMOCYCLIC AND PHOTOCYCLIC ENTRAINMENT OF CIRCADIAN LOCOMOTOR ACTIVITY RHYTHMS IN SLEEPY LIZARDS,TILIQUA RUGOSA

Australian sleepy lizards (Tiliqua rugosa) exhibit marked locomotor activity rhythms in the field and laboratory. Light-dark (LD) and temperature cycles (TCs) are considered important for the entrainment of circadian locomotor activity rhythms and for mediating seasonal adjustments in aspects of these rhythms, such as phase, amplitude, and activity pattern. The relative importance of 24 h LD and TCs in entraining the circadian locomotor activity rhythm in T. rugosa was examined in three experiments. In the first experiment, lizards were held under LD 12:12 and subjected to either a TC of 33:15?°?C in phase with the LD cycle or a reversed TC positioned in antiphase to the LD cycle. Following LD 12:12, lizards were maintained under the same TCs but were subjected to DD. Activity was restricted to the thermophase in LD, irrespective of the lighting regime and during the period of DD that followed, suggesting entrainment by the TC. The amplitude of the TC was lowered by 8?°?C to reduce the intensity and possible masking effect of the TC zeitgeber in subsequent experiments. In the second experiment, lizards were held under LD 12.5:11.5 and subjected to one of three treatments: constant 30?°?C, normal TC (30:20?°?C) in phase with the LD cycle, or reversed TC. Following LD, all lizards were subjected to DD and constant 30?°?C. Post-entrainment free-run records revealed that LD cycles and TCs could both entrain the locomotor rhythms of T. rugosa. In LD, mean activity duration (α) of lizards in the normal TC group was considerably less than that in the constant 30?°?C group. Mean α also increased between LD and DD in lizards in the normal TC group. Although there was large variation in the phasing of the rhythm in relation to the LD cycle in reversed TC lizards, TCs presented in phase with the LD cycle most accurately synchronized the rhythm to the photocycle. In the third experiment, lizards were held in DD at constant 30?°?C before being subjected to a further period of DD and one of four treatments: normal TC (06:00 to 18:00 h thermophase), delayed TC (12:00 to 00:00 h thermophase), advanced TC (00:00 to 12:00 h thermophase), or control (no TC, constant 30?°?C). While control lizards continued to free-run in DD at constant temperature, the locomotor activity rhythms of lizards subjected to TCs rapidly entrained to TCs, whether or not the TC was phase advanced or delayed by 6 h. There was no difference in the phase relationships of lizard activity rhythms to the onset of the thermophase among the normal, delayed, and advanced TC groups, suggesting equally strong entrainment to the TC in each group. The results of this experiment excluded the possibility that masking effects were responsible for the locomotor activity responses of lizards to TCs. The three experiments demonstrated that TCs are important for entraining circadian locomotor activity rhythms of T. rugosa, even when photic cues are conflicting or absent, and that an interaction between LD cycles and TCs most accurately synchronizes this rhythm. (Author correspondence: david.j.ellis@adelaide.edu.au)     

Aggressiveness during monogamous pairing in the sleepy lizard, <Emphasis Type="Italic">Tiliqua rugosa</Emphasis>: a test of the mate guarding hypothesis

The Australian sleepy lizard, Tiliqua rugosa, maintains monogamous associations for an average of 6 weeks before mating each spring. One hypothesis to explain this prolonged partnership is that males are guarding their female partners from rival males. This hypothesis has three predictions, that males are more aggressive than females to conspecific males, that male aggression will increase as the time of mating gets closer, and that males will be more aggressive towards conspecific males when they are with their partner than when they are alone. We tested those predictions with indirect evidence of aggression, using counts of scale damage on randomly encountered lizards, and with direct observations of their responses to approaches by conspecific and heterospecific models. As predicted by the mate guarding hypothesis, males showed more evidence of aggression towards conspecifics than did females. However, in contrast to the hypothesis, males did not become more aggressive as the time of mating came closer, and males in pairs were less aggressive than males on their own. Mate guarding cannot be the only process that has led to the prolonged monogamous associations in this species. Parental care is also unknown in these lizards, and we suggest that monogamy may be maintained through some form of female coercion, allowing females to gain additional fitness from the enhanced vigilance that results from male proximity.     

The visual perceptual range of a lizard, <Emphasis Type="Italic">Tiliqua rugosa</Emphasis>

The fragmentation of landscapes produces habitat gaps where the distance between visual landmarks may exceed the perceptual range of a species and impose navigational constraints. We estimated the visual perceptual range of the Australian sleepy lizard, Tiliqua rugosa, by releasing individuals in the centre of a cleared arena in high temperature conditions, with a 0.5-m-high bush placed either 10, 20 or 30 m from the release site. Lizards were more likely to locate those bushes and shelter under them when they were closer, and no lizards found a bush at 30 m. In addition, lizards were less likely to move from the release point when bushes were at 30 m than when they were at the two closer distances. These data suggest that for sleepy lizards the perceptual range for a 0.5-m-high bush is about 20 m. In the uncleared chenopod shrub-land where these lizards live, suitable shelter bushes are an average of 10.5 m from any point in their home range, well within their perceptual range.     

Ecological aspects of thermoregulation at high altitudes: the case of andean Liolaemus lizards in northern Chile

Summary We document activity field temperatures, daily activity patterns, and extent of thermoregulation in four species of Liolaemus lizards inhabiting at high altitude (above 3500 m) in the Andes of northern Chile. These four species have similar activity field temperature (Tb near 29°C) despite their being distributed at different altitudinal belts. However, conspicuous differences exist between higher-altitude (L. alticolor and L. jamesi) and lower-altitude (L. islugensis and L. ornatus) lizards regarding extent of thermoregulation and activity period. Some differences in morphology, behavior, and patterns of microhabitat occupancy are also apparent among these four species and are seemingly related to the thermal environment to which they are subjected. In comparison to eight low-altitude Liolaemus species in central Chile (Tb near 35°C) the four high-altitude species in northern Chile have lower activity field temperature. The latter is apparently due to the constraints imposed by the harsh Andean thermal environment, a hypothesis supported by the fact that high-altitude Liolaemus lizards under laboratory conditions demonstrate body temperatures that exceed by 5°C or more, those recorded in the field.     

Thyroxine affects behavioral thermoregulation but not growth rate among populations of the western fence lizard (Sceloporus occidentalis)

We investigated whether thyroxine influences hatchling growth rate of the western fence lizard (Sceloporus occidentalis) throught its effects on thermoregulatory behaviors. We reared control and thyroxine-injected hatchlings from three populations of S. occidentalis that differ in growth rate in a thermal gradient. We also measured the daily changes in body temperature and activity level (proportion of time spent out of retreat sites) of control and thyroxine-injected lizards. Previous studies have shown that within and among population differences in growth rate of the western fence lizard are correlated with the maintenance of high activity levels (proportion of time spent outside of retreat sites) and high body temperatures throughout the day. Growth rate was not influenced by injections of thyroxine. However, injections of thyroxine did elevate average daily body temperature and daily activity. Although administration of thyroxine uniformly increased the probability of activity throughout the day, it did not appear to alter the daily changes in activity. Previous studies have shown that the slower-growing hatchlings from northern populations exhibit a decline in activity during the later part of the thermal cycle, whereas the faster growing southern hatchlings maintain the same level of high activity throughout the thermal cycle. The decline in activity of northern populations was not prevented by thyroxine injection used in our current study. Northern lizards receiving exogenous thyroxine were still less active later in the day compared to early in the day, even though activity level throughout the day was elevated. Thus, the effects of thyroxine on temperature regulation observed in our study (general increase in activity level) appear to be unrelated to those aspects of temperature regulation (e.g., daily changes in behavioral thermoregulation) that are correlated with among population differences in growth rate. We also raised hatchlings in a cycling thermal regime (forced thermal cycle of 34°C:15°C, 12L:12D) where behavioral thermoregulation is not possible. The growth rate of lizards forced to cycle between 34°C:15°C on a daily basis was significantly lower than those lizards allowed to behaviorally thermoregulate, further underscoring the importance of the circadian pattern of thermoregulation for growth.Abbreviations GR growth rate - MR metabolic rate - SMR standard metabolic rate - SVL snout-vent length - T₄ thyroxine - T _b body temperature - T _e environmental temperature     

<Emphasis Type="Italic">Candida rugosa</Emphasis> lipase-catalyzed polyurethane degradation in aqueous medium

Candida rugosa lipase (EC 3.1.1.3) was used to degrade commercially-available solid poly(ester)urethane (Impranil) in an aqueous medium under different temperature, pH, enzyme and substrate concentrations. A mathematical model was developed and applied to represent the degradation kinetics of the solid polyurethane. Reaction optima were found to be pH 7 and 35°C. Diethylene glycol, a degradation byproduct, generation rate was measured to be 0.12 mg/l min and the activation energy was calculated as 9.121 kcal/gmol K. This information will be useful in developing bioreactors for practical applications to manage polyurethane wastes using lipase.     

Rational strategy for the production of new crude lipases from <Emphasis Type="BoldItalic">Candida rugosa</Emphasis>

Candida rugosa was cultured using different inducers (oleic acid, olive oil, sunflower oil, n-dodecanol and glycerol) as the only carbon source in batch conditions, as well as in several fed-batch fermentations (oleic acid as inducer) at variable feed rate conditions. The N-terminal analysis of each crude lipase revealed that, while the isoenzymes Lip2 and Lip3 are always secreted (at different proportions depending on the inducer), Lip1 was produced only using n-dodecanol (batch conditions) or oleic acid (fed-batch at high feed rate). The nature of the inducer controls the isoenzyme percentage; when this is fixed, as well as the feed rate in fed-batch fermentation, the isoenzymatic profile remained unaltered and the samples differed only in the activity of the lipases, as determined by heptyl oleate synthesis.     

Turtles (<Emphasis Type="Italic">Chelodina longicollis</Emphasis>) regulate muscle metabolic enzyme activity in response to seasonal variation in body temperature

Fluctuations in the thermal environment may elicit different responses in animals: migration to climatically different areas, regulation of body temperature, modification of biochemical reaction rates, or assuming a state of dormancy. Many ectothermic reptiles are active over a range of body temperatures that vary seasonally. Here we test the hypothesis that metabolic enzyme activity acclimatises seasonally in freshwater turtles (Chelodina longicollis) in addition to, or instead of, behavioural regulation of body temperatures. We measured body temperatures in free-ranging turtles (n=3) by radiotelemetry, and we assayed phosphofructokinase (PFK), lactate dehydrogenase (LDH), citrate synthase (CS) and cytochrome c oxidase (CCO) activities in early autumn (March, n=10 turtles), late autumn (May, n=7) and mid-winter (July, n=7) over a range of assay temperatures (10 °C, 15 °C, 20 °C, 25 °C). Body temperatures were either not different from, or higher than expected from a theoretical null-distribution of a randomly moving animal. Field body temperatures at any season were lower, however, than expected from animals that maximised their sun exposure. Turtles maintained constant PFK, LDH and CCO activities in different months, despite body temperature differences of nearly 13.0 °C between March (average daily body temperature=24.4 °C) and July (average=11.4 °C). CS activity did not vary between March and May (average daily body temperature=20.2 °C), but it decreased in July. Thus C. longicollis use a combination of behavioural thermoregulation and biochemical acclimatisation in response to seasonally changing thermal conditions. Ectothermic reptiles were often thought not to acclimatise biochemically, and our results show that behavioural attainment of a preferred body temperature is not mandatory for activity or physiological performance in turtles.Abbreviations CS citrate synthase - CCO cytochrome c oxidase - LDH lactate dehydrogenase - PFK phosphofructokinase Communicated by I.D. Hume     

野生玫瑰种群表型变异

在野生玫瑰(Rosa rugosa)自然分布区内选取5个代表种群,选择24个表型性状作为研究对象,运用方差分析、多重比较、主成分分析、相关分析、聚类分析等方法,得出野生玫瑰种群表型变异程度和变异规律。结果显示:(1)24个表型性状在种群间和种群内均存在极显著差异,变异非常丰富;种群内变异(0.2718)大于种群间变异(0.1679),种群内变异是表型变异的主要来源,种群间平均表型分化系数(VST)为0.2952,分化水平相对较大;表型性状平均变异系数(CV)为18.48%(6.67—26.79%),叶片、果实、花、种子的变异系数依次为21.40%、17.42%、12.54%、6.67%;主成分分析表明叶片、果实的表型变异对种群变异起主要的贡献作用。(2)托叶长与年平均气温、7月平均气温呈显著正相关,与经、纬度呈显著负相关;果实横径与年降水量呈显著正相关;千粒重与经、纬度呈显著正相关,与7月平均气温呈显著负相关。(3)利用欧氏距离进行系统聚类分析,可以将5个种群划分为3类,表型性状主要依地理位置聚类。     

5个不同产地藿香叶片解剖结构与光合特性比较研究

以5个不同产地的藿香为研究对象,通过比较叶片解剖结构、光合特性,为藿香在引进地的栽培技术制定提供理论依据。结果显示:(1)不同产地藿香叶片栅栏组织均由1列长柱形细胞构成,其叶片海绵组织细胞在4~6层之间,并在叶片下表皮有较多的气孔和非腺毛,叶片气孔多为卵圆形;其中,成都藿香气孔密度、直径最大,非腺毛长度最长,且叶绿素含量明显高于其它产地。(2)不同产地藿香的光饱和点(LSP)和光补偿点(LCP)分别在688.67~1 244.56和19.42~73.62μmol·m~(-2)·s~(-1)之间,表观量子效率(AQY)则为商丘藿香最高。(3)5个产地的藿香光合日变化曲线均呈双峰型,两个净光合速率(Pn)峰值点均出现在10:00和16:00左右,14:00左右均出现光合午休现象,且均由气孔限制因素导致。研究表明,各产地间藿香叶片解剖结构存在明显差异,并在部分产地中,藿香叶片形态结构与光响应特性存在一定的相关性;成都藿香表现出较强的光环境适应性和光合能力,可作为当地引种优先考虑对象。     

Somatic embryogenesis and plant regeneration in zygotic embryo explant cultures of rugosa rose

Rugosa rose (Rosa rugosa) is cultivated as a garden flower and an important genetic resource for the breeding of roses (R. hybrida). This study describes culture conditions for high frequency plant regeneration from zygotic embryo explants via somatic embryogenesis in rugosa rose. Mature zygotic embryo, cotyledon, and radicle explants formed embryogenic calluses at frequencies of 38, 6.7, and 8.8% when cultured on half-strength Murashige and Skoog medium (½MS) supplemented with 2.26, 9.05, and 9.05 μM 2,4-dichlorophenoxyacetic acid, respectively. Embryogenic calluses produced numerous somatic embryos, which then developed into plantlets on ½MS without growth regulators. Regenerated plantlets were grown to whole plants in a growth chamber.     

Thermal effects on activity patterns and behavioural switching in a concourse of foragers on Stachytarpheta mutabilis (Verbenaceae) in Papua New Guinea

Summary 1. The foraging activities of the papilionid butterflies Ornithoptera priamus poseidon and Papilio ulysses, and the solitary bee Amegilla sapiens (Apoidea, Anthophoridae) on the shrub Stachytarpheta mutabilis were studied in highland Papua New Guinea. 2. The insects' activity patterns were analysed at three sites with differing diurnal microclimate variation. O. priamus and A. sapiens foraged in the morning (after a period of basking and wing-whirring) and late afternoon when temperatures were well below daily maxima, whereas P. ulysses showed foraging peaks during the hottest part of the day. 3. Site choice by all 3 species appeared to be determined primarily by temperature, but within the limits imposed by temperature, nectar supplies probably determined which site was visited. 4. P. ulysses showed interspersed foraging and courtship behaviour, and no behavioural switching was observed for this species. At high temperatures, both O. priamus and A. sapiens ceased foraging and showed territorial and courtship behaviour. This behavioural change allowed avoidance of heat stress, and occurred even when nectar supplies were maintained at high levels. 5. Thermal effects on behavioural switching in these insects are compared with related phenomena in other bees and butterflies.     

Feeding on seeds induces diapause in the cabbage bug, Eurydema rugosa

Eurydema rugosa Motschulsky (Heteroptera: Pentatomidae) entered adult diapause under a short-day photoperiod, whether the food was leaves or seeds of Brassica napus L. Under a long-day photoperiod, adults raised on leaves became reproductive. However, adults raised on seeds entered diapause even under a long-day photoperiod, although they were nutritionally better than leaves in respect of mortality and weight gain in the nymphal period. It is suggested that E. rugosa uses the status of the host plant as a cue for timing the diapause induction.

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Résumé E. rugosa Motschulsky (Hétérop. Pentatomidae) entre en diapause imaginale en jours courts, lorsqu'il s'alimente avec des feuilles ou des graines de Brassica napus L. En jours longs, les adultes élevés sur feuilles se reproduisent; par contre, élevés sur graines, ils entrent en diapause, bien que leur alimentation ait été meilleure que sur feuilles à en juger par la mortalité et le gain de poids pendant le dernier stade larvaire. On suggère que E. rugosa utilise l'état de la plante hôte comme moyen de régler l'induction de la diapause.

     

High lipase production by Candida rugosa is associated with G1 cells. A flow cytometry study

Growth of C. rugosa on three different culture media was analysed by laser flow cytometry to evaluate physiological growth conditions allowing effective lipase production. The highest productivity was associated with an increased proportion of cells in the G1 phase and was independent of the effect of the medium on lipase formation.     

High-yield synthesis of wax esters catalysed by modified <Emphasis Type="Italic">Candida rugosa</Emphasis> lipase

Candida rugosa lipase (CRL) was applied in a non-solvent esterification reaction to yield twelve wax esters. All products were obtained in nearly 100% yield for 10 h at 50°C when immobilized PEG₂₀₀₀-activated C. rugosa lipase was added to the reaction mixture. The surfactant had also a beneficial effect on the stability of the biocatalytic preparation with 83% of its activity conserved after the seventh run of repeated batch reactions.     

Respiratory cooling in rattlesnakes

We used infrared thermography to study respiratory cooling in the rattlesnakes (Viperidae: Crotalinae) and to partition the effects of air temperature, humidity, and activity levels on head-body temperature differences. We observed a single, cooled region centered around the mouth and nasal capsule that extended across the pit membrane at air temperatures above 20 degrees C. Both head and body temperatures of rattlesnakes increased linearly with air temperature. Head-body temperature differentials also increased with air temperature, but declined significantly at higher relative humidities. Rattling rattlesnakes exhibited significantly greater head-body temperature differentials than did resting rattlesnakes. We suggest that respiratory cooling may provide a thermal buffer for the thermoreceptive pit organs at high air temperatures, but caution that this adaptive hypothesis must be tested with direct neural or behavioral assays.     

Rosmarinic acid production in hairy root cultures of Agastache rugosa Kuntze

Rosmarinic acid, an important phenolic active compound, is one of the main active constituents of Agastache rugosa Kuntze and has astringent properties, antioxidant capacity, anti-inflammatory activity, antimutagenic ability, antimicrobial capacity, and antiviral properties. To investigate in vitro production of rosmarinic acid, we established a hairy root culture of A. rugosa by infecting leaf and stem explants with Agrobacterium rhizogenes R1000, and tested the growth and rosmarinic acid production of these cultures. Hairy roots were cultured in Murashige and Skoog liquid medium and maximum growth (14.1 g dry wt/l) was attained after 14 days of culture, at which time the content of rosmarinic acid was 116 mg/g dry wt. The present results demonstrate that hairy root culture of A. rugosa is a valuable alternative approach for the production of rosmarinic acid.