福寿螺的过冷却研究

福寿螺是IUCN认定的世界100种恶性外来入侵物种之一,在华南地区已对水稻生产造成严重危害。福寿螺属于热带软体动物,采用万用电表联接热敏电阻法研究了福寿螺在低温胁迫下的过冷却点,并探讨了不同个体福寿螺在过冷却发生后的死亡率和体内组织损伤。结果表明: 1)各种螺高的福寿螺冷却点均值为-6.96 ℃,范围在-6.21--7.32 ℃之间,恢复最高体温均值为-4.07 ℃,范围在-3.07--4.93 ℃之间,过冷却后维持时间均值为45.97 min,范围在18.60-75.34 min;2)福寿螺的过冷却点大小受螺高影响,螺高35 mm≤H<45 mm的福寿螺过冷却点显著高于5 mm≤H<15 mm、15 mm≤H<25 mm、25 mm≤H<35 mm体型的福寿螺,5-35 mm螺高的福寿螺过冷却点变化稳定;3)福寿螺在发生过冷却阶段的死亡率在23.33%-36.67%之间,不同体型的福寿螺之间没有显著性差异;4)过冷却后福寿螺死亡率随暴露时间的延长而提高,0到15 min内由20.9%提高到100%,过冷却后暴露时间对福寿螺死亡率影响较大;5)发生过冷却15 min后取出的福寿螺,染色后其外套膜有少量红色出现,而肾部和消化腺部未呈现明显红色,低温胁迫已经对福寿螺组织造成了显著损伤。本研究结果对于进一步探索福寿螺的生态适应性、扩散北界及福寿螺的越冬机制有一定参考价值。     

Influence of soil moisture on egg cold hardiness in the migratory locust Locusta migratoria (Orthoptera: Acridiidae)

Abstract.  The present study investigates the influence of environmental moisture on cold hardiness of the migratory locust, Locusta migratoria . The water content of locust eggs kept in soil at 30 °C varies according to the moisture content of the substrate. In turn, it can significantly affect the supercooling point of locust eggs (range from −26 to −14.8 °C) and the mortality when exposed to subzero temperatures. Environmental moisture influences the supercooling capacity of eggs and their survival at low temperature. When locust eggs of the same water content are exposed to subzero temperatures under different soil moistures, their mortality varies between short-time exposure and long-time exposure at subzero temperatures. Given a short-time exposure, mortality in wet soil is lower than in dry soil due to the buffering effect of soil water against temperature change. The pattern of egg mortality is reversed after long-time exposure at low temperature, suggesting that inoculative freezing may be an important mortality factor. It is suggested that interactions between soil moisture and low temperature can influence the cold hardiness of locust eggs, and partial dehydration is beneficial to over-wintering eggs of the migratory locust.     

Cold hardiness in the black rice bug, Scotinophara lurida

Abstract.  The mechanisms and strategies for winter survival of the black rice bug Scotinophara lurida are investigated along with the relationship between cold hardiness and diapause. The ability of S. lurida to survive subzero temperatures varies depending on developmental stage, temperature and exposure duration. Mean supercooling point (SCP) varies from –7.6 to –10.7 °C with developmental stage, but is not significantly different between stages examined. The SCP also varies with season, being lowest in January and increasing rapidly in February and remaining almost at the same level (–7.3 to 9.6 °C) until April The osmolality of haemolymph of field-collected S. lurida adults rises dramatically from 53.9 mOsm kg⁻¹ in August to 75.3 mOsm kg⁻¹ in December, and then declines linearly to 57.0 mOsm kg⁻¹ in May. Field-collected S. lurida adults show a peak glucose content in October, glycerol content in November and trehalose content in December. Only trehalose content decreases after the application of the juvenile hormone analogue, fenoxycarb, suggesting that trehalose is a cryoprotectant during diapause. These various physiological and biochemical traits related to cold tolerance in S. lurida may be, at least in part, under the control of juvenile hormone through the reproductive diapause programme.     

Cold hardiness of Habrobracon hebetor (Say) (Hymenoptera: Braconidae), a parasitoid of pyralid moths

The ectoparasitoid Habrobracon hebetor (Say) attacks stored-product infesting pyralid moths that are able to overwinter under extremely cold conditions. The extent to which H. hebetor can withstand these conditions is not known, but has important implications for the ability of H. hebetor to provide long-term suppression of these pests in temperate climates. We investigated basic cold hardiness aspects of a mutant eye-color strain of H. hebetor. Feeding larvae and adults of H. hebetor had supercooling points (SCPs) at temperatures higher than those of eggs and pupae. Mean SCPs of females and males were equivalent, as were those of naked and silk-encased pupae. Feeding on honey prior to being subjected to low temperatures significantly increased the SCP of adult females by approximately 8 degrees C. Mortality of pupae and adults increased significantly whenever the temperature dropped below the mean SCP, indicating that H. hebetor does not tolerate freezing. For pupae and adults exposed to -12 and -5 degrees C, the hourly mortality rate increased with time of exposure. Pupae and adults exposed to -12 degrees C for different time intervals showed high mortality after only 1d of exposure. At -5 degrees C, none survived 12d of exposure. A better understanding of how well this parasitoid tolerates low temperatures will be useful in evaluating its potential as a biological control agent of stored-product moths in temperate regions.     

Cold hardiness in the Alpine beetles Patrobus septentrionis and Calathus melanocephalus

Alpine Patrobus septentrionis and Calathus melanocephalus (Col., Carabidae) were found to be susceptible to freezing. In the summer, the supercooling points were about ?5 to ?6°C. They were lowered during acclimation in the field and in the laboratory at 0 or ?3°C. Cold hardiness was correlated to, but not determined by, haemolymph osmolality. Thermal hysteresis was not detected. Increase in cold hardiness was concluded to be mainly a result of the influence of acclimation conditions on ice-nucleating compounds. In P. septentrionis, the results indicate that different compounds with ice-nucleation activity at different temperatures determine the limit of supercooling at different times. The haemolymph of both species supercooled well below the intact beetles at all seasons. Changes in haemolymph supercooling points could be ascribed to inactivation of ice nucleators in early autumn and to the effect of changes in solute concentration. In P. septentrionis, myo-inositol increased during cold-acclimation to 80–120 mMol concentrations, whereas C. melanocephalus produced 40–60 mMol trehalose.     

Cold tolerance of first-instar nymphs of the Australian plague locust, Chortoicetes terminifera

The cold tolerance of first-instar nymphs of the Australian plague locust, Chortoicetes terminifera, was examined using measures of total body water content, supercooling point and mortality for a range of sub-zero temperature exposure regimes. The supercooling points for starved and fed nymphs were −13.1 ± 0.9 and −12.6 ± 1.6 °C, and freezing caused complete mortality. Above these temperatures, nymphs were cold tolerant to different degrees based on whether they were starved or given access to food and water for 24 h prior to exposure. The rate of cooling also had a significant effect on mortality. Very rapid cooling to −7 °C caused 84 and 87% mortality for starved and fed nymphs respectively, but this significantly decreased for starved nymphs if temperature declined by more ecologically realistic rates of 0.5 and 0.1 °C min⁻¹. These results are indicative of a rapid cold hardening response and are discussed in terms of the likely effects of cold nights and frost on first-instar nymphal survival in the field.     

Moulting reduces freeze susceptibility in the Antarctic mite Alaskozetes antarcticus (Michael)

Abstract.  The effect of moulting on the cold hardiness of the oribatid mite Alaskozetes antarcticus (Michael) is investigated. Non moulting animals show clear seasonal patterns of cold hardiness with high supercooling points (SCPs) at the peak of summer and an increasing proportion of low SCPs with declining environmental temperatures. By contrast, both field-fresh and laboratory acclimated (5 °C) mites in the moult state are consistently found to have low SCPs regardless of environmental temperature.     

桔小实蝇不同发育阶段过冷却点的测定

对桔小实蝇Bactrocera dorsalis (Hendel)不同发育阶段的过冷却点进行了测定。结果表明：同一虫期个体间的过冷却点出现不同程度的变异,但均服从正态分布。不同虫期的过冷却点差异显著,其中蛹的过冷却点最低（-12.2℃～-15.0℃）。老熟幼虫的过冷却点为-8.1℃。成虫的过冷却点最低值为7日龄雄虫（-10.5℃）和雌虫（-10.1℃）,最高值为60日龄雄虫（-5.9℃）和雌虫（-6.4℃）,但同一发育时期的雌、雄成虫之间的过冷却点没有差异。测定结果提示蛹期最有可能是该虫在温带地区越冬的虫态。     

上海地区红棕象甲的耐寒性研究

红棕象甲Rhynchophorus ferrugineus (Olivier)是我国危害棕榈科植物的重要入侵害虫, 为探明其越冬抗寒性, 利用过冷却点测定仪测定了红棕象甲上海种群不同虫态的过冷却点, 在低温箱内测定了该虫不同虫态的耐寒性, 然后结合田间越冬模拟试验、气象资料和寄主分布情况, 初步确定了红棕象甲在我国的越冬北界。结果表明： 红棕象甲过冷却点随虫态的发育程度的升高而下降, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫的平均过冷却点分别为-5.92, -6.42, -7.19, -7.43和-11.84℃, 过冷却点由高到低的顺序依次为: 卵>幼虫>成虫。在6, 24, 48和72 h 4个时间处理下, 各虫态在低温与存活率之间呈显著或极显著的logistic回归, 半致死温度（Ltemp₅₀）均随处理时间的延长而上升, 不同虫态在处理72 h 后, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫之间的Ltemp₅₀分别为1.61, -1.67, -2.39, -2.40和-0.40℃, 各虫态耐寒性由弱到强的顺序依次为: 卵<成虫<幼虫。红棕象甲不同发育阶段的过冷却点与其耐寒性并不完全相关, 幼虫和成虫均可能是该虫的越冬虫态。连续两年的田间模拟越冬试验表明, 在上海地区, 红棕象甲的幼虫和成虫的越冬存活率均在60％以上, 说明红棕象甲在上海地区是可以越冬的。根据这些结果, 结合寄主分布情况, 初步将红棕象甲在我国的越冬北界定于北纬35°附近, 即1月份0℃等温线左右。     

A peculiar new species in the genus Landouria Godwin-Austen, 1918 from China (Gastropoda: Heterobranchia: Stylommatophora: Camaenidae)

Landouria omphalostoma n. sp. is described from northern Yunnan, China. The new species is placed in the genus Landouria Godwin-Austen, 1918 based on the presence of tuberculated flagellum and swollen basal part in the bursa copulatrix, and on the absence of dart sacs and mucous glands. L. omphalostoma n. sp. is characterized by a horizontal aperture of the shell which is unique in the genus. This feature resembles those of only two other camaenid/bradybaenid taxa, species of the Chinese genus Pseudaspasita Möllendorff, 1902 and species of the Japanese genus Coelorus Pilsbry, 1900. This is the first verified record of the genus in China and the easternmost record of Landouria. This paper provides the taxonomic ground work for further studies of the repeated evolution of a horizontal aperture.     

昆虫的耐寒性及其影响因素

昆虫是变温动物,种群的繁衍面临如何安全度过漫长而寒冷的冬季的挑战.为了安全越冬,昆虫必须适应冬季的低温,增强耐寒能力是昆虫的一种重要的季节适应机制.近年来,关于昆虫耐寒性的研究不断开展,研究内容涉及影响耐寒性的因素、耐寒性的机制等方面.影响昆虫耐寒性的因素包括环境因素,如气候的季节变化、纬度或海拔的差异等,以及昆虫自身的发育阶段、滞育发生等.     

Winter survival of an adult bark beetle Ips acuminatus Gyll

Freezing-susceptible adult Ips acuminatus hibernate underneath bark of Scots pine. The beetles lower their supercooling points from ?20 to ?34°C due to accumulation of low molecular weight antifreezes. The capability of specimens to supercool to about ?20°C in the absence of cryoprotective solutes during winter, seemed to be at least partially attributable to the presence of a thermal hysteresis factor at 3–4°C.Using a GC-MS-COM technique, a unique combination of accumulated solutes present only in specimens demonstrating supercooling points below ?20°C was identified as ethylene glycol, mannitol, sorbitol and dulcitol. Not previously found in nature, ethylene glycol was the major solute (90%) synthesized at sub-zero temperatures. Exposure to ?10°C was an effective cue to accumulation of ethylene glycol and nearly 5 times as effective in promoting sorbitol synthesis than was ?5°C. When low molecular weight substances were lost at high temperatures, they were not re-synthesized in beetles re-exposed to sub-zero temperature. The supercooling point was closely related to both the concentration of ethylene glycol and to the haemolymph melting point. Attempts to correlate changes in sorbitol concentrations to changes in supercooling points were not conclusive.Proliferation of thermal hysteresis was observed in the beginning of November. A melting-hysteresis freezing point differential of about 3.6°C was demonstrated in the haemolymph of beetles during December. No thermal hysteresis was demonstrated in the haemolymph of positive phototactic beetles or in the outdoor beetles in May. The combination of high temperature and long photoperiod appeared to be a more effective cue to the final loss of thermal hysteresis than was high temperature alone.     

Radiation of the land snail genus Sagda (Pulmonata: Sagdidae): comparative morphology, biogeography and ecology of the species of north-central Jamaica

The radiation of the endemic Jamaican land snail genus Sagda is examined through analysis of the comparative shell morphology, anatomy, ecology and biogeography of the four species inhabiting north-central Jamaica: S. centralis sp. nov. , S. spei, S. bondi and S. montegoensis. The species are all rather similar in their morphology and ecology. The shells show some differences in the presence or absence of internal lamellae, the position of the lamellae, the form of the depression in the centre of the base, the regularity of sculpture (low ribs) and the pattern of microsculpture. Slight differences in the form of the radular teeth are found between some species. The structure of the secondary ureter is peculiar: in all species it is open shortly behind the mantle collar, then branches into two closed tubes which open separately on the outside of the mantle collar. Despite its great complexity, the reproductive system is rather uniform among species; differences are seen in the number of pockets that form spines on the spermatophore. The complex, three-branched spermatophore is formed within the entire epiphallar branch of the penis, whereas the appendicular branch apparently assumes the copulatory function. When inactive, all species are found associated with rocks or, less commonly, among leaf litter. Activity occurs primarily at night and mostly on leaf litter rather than rocks. The snails eat dead plant material, including a variety of plant species. All species (with the possible exception of S. montegoensis) are ovoviviparous. They overlap broadly in the range of climates occupied. The geographic ranges of the four species are overlapping, although Sagda bondi has a parapatric distribution in relation to the western species S. grandis. Fossil and subfossil evidence shows that shifts in the ranges of the species have occurred in the late Quaternary; Pleistocene-Holocene environmental changes, habitat destruction by man, and possibly other environmental changes during historical times are probably involved. Radiations of other land snail groups on tropical islands have generally produced greater morphological and ecological differentiation than in Sagda. Allopatric or parapatric species distributions appear to be more common in other land snail radiations.     

Cold-hardiness in several species of land snails

1. 1.|Mean supercooling points (SCP) of Anguispira alternata (Say) gradually declined in autumn and increased in spring. SCP also declined in autumn in Discus conrkhitei (Newcomb) and Gastrocopta armifera (Say).

2. 2.|Supercooling ability varied among the species, being the greatest in G. armifera.

3. 3.|Prolonged exposure of winter animals to high temperatures resulted in only a modest increase in SCP.

4. 4.|Low temperature survival of D. cronkhitei and G. armifera was slightly poorer than predicted by the mean SCP.

Establishment of the comet assay in the freshwater snail Biomphalaria glabrata (Say, 1818)

The single cell gel eletrophoresis or the comet assay was established in the freshwater snail Biomphalaria glabrata. For detecting DNA damage in circulating hemocytes, adult snails were irradiated with single doses of 2.5, 5, 10 and 20 Gy of ⁶⁰Co gamma radiation. Genotoxic effect of ionizing radiation was detected at all doses as a dose-related increase in DNA migration. Comet assay in B. glabrata demonstrated to be a simple, fast and reliable tool in the evaluation of genotoxic effects of environmental mutagens.     

Mucus trail following by the snail Biomphalaria glabrata (Say)

An experimental procedure involving time-lapse cinematography was used to investigate the responses of the freshwater pulmonate, Biomphalaria glabrata (Say), to their mucus trails. It has been shown that individuals of this species are capable of following their own trails and also those laid by other individuals of the same species. The capacity of mucus trails to influence snail behaviour is lost after a brief period of between 10 and 30 min. The snails tend to move in the direction the trail was laid with expectation greater than by chance. They do not follow mucus trails laid by another species, Limnaea stagnalis. The possible adaptive significance of trail following is discussed.     

寄主对桔小实蝇耐寒性的影响

为了研究寄主营养对桔小实蝇耐寒性的作用,测定了以15种果蔬饲养的桔小实蝇1日龄蛹的过冷却点(supercooling points,SCP); 再选取南瓜、西红柿、柑桔、番石榴和杨桃等5种果蔬,测定了桔小实蝇3龄老熟幼虫、1日龄蛹、3日龄蛹、5日龄蛹、7日龄蛹和雌雄成虫的过冷却点,并观察了1日龄蛹的低温存活力。结果表明：（1）15种果蔬饲养所得的桔小实蝇1日龄蛹SCP均值在-11.03℃～-13.17℃,不同寄主发育的桔小实蝇SCP值存在显著性差异,其中以取食蒲桃的最高,为-11.03℃,取食苦瓜的最低,为-13.17℃。（2）5种果蔬饲养所得的桔小实蝇各虫态的SCP均值存在极显著差异(F_(4,863)＝35.6,P<0.01); 同一寄主上的桔小实蝇不同虫态之间SCP均值也达到极显著性差异(F_(6,863)＝392.9,P<0.01); 且寄主和发育龄期之间存在着极显著的交互作用(F_(24,863)＝9.4,P<0.01)。（3）桔小实蝇各发育阶段,SCP值表现一定变化： 老熟幼虫发育至1日龄蛹,SCP值变化不大; 蛹发育至3、5和7天过冷却能力明显增强,降至-20℃左右,但他们之间没有明显区别; 羽化后3～5天的成虫SCP值又升高至-10℃左右。老熟幼虫、1日龄蛹和2～3日龄成虫与3日龄、5日龄和7日龄蛹的SCP值之间有显著性差异。（4）将5种果蔬饲养所得的桔小实蝇1日龄蛹置于6℃和-3℃下进行较长时间(1～8天)和较短时间(1～8 h)的低温处理,发现番石榴、杨桃和南瓜发育的蛹经低温处理后的校正羽化率较西红柿和柑桔发育的蛹高; 同样在0℃、3℃、6℃和9℃处理(2天)的实验中,得出相似的结果。因此,本实验结果表明桔小实蝇幼虫由于生活寄主的不同使得其下一代蛹的耐寒性产生了差异,引起其差异的原因值得进一步研究。     

Supercooling ability is surprisingly invariable in eggs of the land snail Cantareus aspersus

From an ontogenetic point of view, invertebrate eggs are generally the most freezing intolerant stage of a species. Development state, water content and acclimation may affect their supercooling ability. In this study, we measured fresh mass, water content and temperature of crystallisation (T(c)) of eggs of the edible land snail Cantareus aspersus, depending on its form ("aspersa"vs. "maxima"), incubation temperature (20, 12 and 7 degrees C) and physiological age (as part of the complete development). We also tested their tolerance to freezing. Despite a high number of individual observations (n=759) and significant differences of fresh mass and water content between both subspecies, no effect of origin, incubation temperature or development state has been found in this study. T(c) remained constant whatever the condition, with an overall mean of -5.40+/-0.24 degrees C (mean+/-SD). We suggest that fresh mass is important, a high water content and a constantly wet surface confer to land snail eggs a poor ability to supercool. Moreover, the presence of ice nucleating agents at the egg surface (microorganisms present in the soil, calcium carbonate crystals of the egg shell) might induce freezing. Thus, considering the present results, to delay hatching by cryopreservation of eggs does not seem possible.     

Cold tolerance and supercooling points of two ladybird beetles (Col.: Coccinellidae): Impact of the diet

Ladybird beetles have successfully been used to control different pests. Cheilomenes sexmaculata (Fabricius) and Oenopia conglobata contaminata)Menetries((Coleoptera: Coccinellidae) are two dominant and efficient predators of the common pistachio psylla, Agonoscena pistaciae (Hem.: Psyllidae) in Iran. In the current study, the impact of two diets, i.e., nymphs of A. pistaciae and eggs of Ephestia kuehniella Zeller (Lep.: Pyralidae), were investigated on the cold hardiness, supercooling point (SCP), and lethal temperature of different life stages of the coccinellids. The results suggested that the eggs of E. kuehniella are a suitable diet for both predators. In general, beetles of O. conglobata contaminata were more cold tolerant than those of C. sexmaculata. The SCP of the adults of C. sexmaculata, feeding on psyllid was about −15 °C, whereas the SCP of the beetles, feeding on the eggs of flour moth, was about −19 °C. However, the diets had no significant effects on the SCP of O. conglobata contaminata. For both ladybird beetles, eggs were the most sensitive stage, and adulthood was the most tolerant developmental stage. No eggs survived at zero and subzero temperatures. The survival of C. sexmaculata increased from 6.25% for adults fed on psyllids to 13.75% for those fed on flour moth eggs after 24 h exposure to cold at −4 °C. The survival of O. conglobata contaminata adults after exposure to cold at −4 °C for 24 h raised from 28.75% for adults fed on psyllids to 42.50% for those fed on flour moth eggs. Regardless of the diet, both beetles were considered to be chill-intolerant insects as the most mortality occurred above the SCP.     

Characterization of 17 microsatellite loci in the Japanese land snail genera Mandarina,Ainohelix, and Euhadra (Mollusca,Gastropoda, Pulmonata)

Land snails of the genera Mandarina, Euhadra and Ainohelix are useful for understanding the ecology and evolution of speciation and adaptation, so we have developed 17 microsatellite loci for these species. As in other land snails, most of the loci are highly polymorphic compound repeats, with a great size range between alleles. The loci should be useful in understanding gene‐flow, genetic structure and speciation in these species.