Dual strategy for immune defense in the land snail Cornu aspersum (Gastropoda, Pulmonata)

Immune defenses have been shown to be heavily involved in the evolution of physiological trade-offs. In this study, we compared the internal defense systems in two subspecies of the land snail Cornu aspersum that exhibit contrasting life-history strategies. The "fast-living" Cornu aspersum subsp. aspersa is widespread throughout the world, especially in ecosystems disturbed by man, whereas natural populations of the giant Cornu aspersum subsp. maxima, characterized by a longer life span, are present only in north Africa. Snails were experimentally challenged with Escherichia coli; the measurements used to assess their internal defense for cell- and humoral-mediated immune responses were bacterial clearance, hemocyte density, reactive oxygen species (ROS) production, and plasma antibacterial activity. Both subspecies showed a similar ability to clear bacteria from their hemolymph; however, they varied in the robustness of different individual immune components. Cornu aspersum aspersa had higher ROS activity than did C. a. maxima and lower plasma bactericidal activity. These results suggest that ecological factors can sculpt the immune response. One interpretation is that shorter life span selects for immune defenses such as ROS that, although effective, can cause long-term damage. Such different immune patterns obviously entail various costs involved in the strong intraspecific variation of life-history trade-offs we previously observed. We also have to consider that such variation might be related to intraspecific differences in the relative strength of resistance and tolerance mechanisms.     

Influence of temperature acclimation and gut content on the supercooling ability of the land snail Cornu aspersum

The invasive land snail Cornu aspersum possesses a low ability to supercool (c. -5 degrees C in winter) and survives only minimal ice formation in its body fluids, what may limit its expansion to colder environments. In the present study, we investigated the influence of acclimation and starvation on its supercooling ability. During eight weeks, individuals were maintained at 20 degrees C, fed or starved, or placed at 5 degrees C, directly or with progressive acclimation to cold and shorter photoperiod. Temperature of crystallisation of whole individual (Tc(I)) and hemolymph (Tc(H)), mass data and gut content were recorded every two weeks. Hemolymphatic glucose and glycerol were measured at the end of experiment and occurrence of intestinal ice-nucleating agents (INA) was researched. Acclimation had no effect on Tc(I) but stimulated purging of the gut. Starvation induced a slight decrease of Tc(I) whereas a high quantity of alimentary particles in the digestive tract limited the supercooling ability. Glucose and glycerol were not synthesized in cold conditions. Mean Tc(H) was low (c. -17 degrees C), some INA being present in hemolymph of fed animals. Intestinal content of starved individuals exhibited a mean Tc of c. -6 degrees C, decreasing to c. -12 degrees after heating, suggesting the presence of organic INA.     

Shell shape of the land snail Cornu aspersum in North Africa: unexpected evidence of a phylogeographical splitting

Anatomical and molecular characters used to differentiate populations of the land snail Cornu aspersum (Helix aspersa) exhibit, in the western Mediterranean, definite and concordant patterns of correlation with geography. Scenarios involving Pliocene geological changes and postglacial expansion during the Pleistocene were proposed in previous studies to account for the establishment of this geographical structure. In the present work, we have performed a spatial analysis of variation in shell morphometrics, after the partitioning of the overall variation into size and shape components by means of a principal component-based approach (Cadima and Jolliffe, 1996). In order to know if the same historical events have also structured shell variation, the analysis includes all the populations from North Africa which were investigated for anatomical and molecular surveys. Contrary to shell size, which shows a significant spatial heterogeneity essentially related to environmental pressures, variation in shell shape components splits the populations according to a geographical pattern reflective of hypotheses suggested for molecular markers and genital anatomy. This implies that the selective forces often invoked to explain spatial changes in shell shape are not the deciding factors in the present case. Moreover, within each of the two geographical clusters defined, Mantel correlograms show that the similarity between populations declines according to an isolation by distance model. Because of the different allometric relationships between shell size and genitalia measurements in Western and Eastern entities of North Africa, mechanical constraints, possibly leading to a precopulatory isolation in the contact zone, are involved.     

Variation in spermathecal morphology is independent of sperm competition intensity in populations of the simultaneously hermaphroditic land snail Cornu aspersum

The complexity of the sperm-storing organ (spermatheca) has been hypothesized to reflect sperm competition intensity in several gastropod species. Furthermore, considerable variation in spermathecal morphology has been detected among populations of the same species. The morphological variation of the fertilization pouch was studied in five populations of the simultaneously hermaphroditic land snail Cornu aspersum (formerly, Helix aspersa). The populations studied differed in snail density and habitat humidity regimes, thus in sperm competition intensity. The study was conducted on wild adult snails and their progeny, which was reared in the laboratory for two successive generations. Finally, the morphology of the spermatheca was correlated to behavioral mating traits of the snails. The fertilization pouch consisted of a simple fertilization chamber and 4-19 blind tubules. The five studied populations did not differ in either mean number of spermathecal tubules, length of the fertilization chamber, length of the main tubule, or cumulative length of all tubules, while they differed in copulation frequency and mating propensity. No correlation was found between snail size and number of tubules, or length of any spermathecal structure measured. Additionally, no correlation was found between any behavioral trait and the morphological characteristics of the spermatheca. Strong correlations were found only among measurements of some of the spermathecal structures. Our results suggest that the complexity of the spermatheca is not related to sperm competition intensity and its structure is thus genetically determined.     

Tracing the Invasion of the Mediterranean Land Snail Cornu aspersum aspersum Becoming an Agricultural and Garden Pest in Areas Recently Introduced

This study is the first on the genetics of invasive populations of one of the most widely spread land mollusc species known in the world, the “Brown Snail” Cornu aspersum aspersum. Deliberately or accidentally imported, the species has become recently a notorious pest outside its native Mediterranean range. We compared the spatial structure and genetic variability of invasive (America, Oceania, South Africa) versus native populations using five microsatellite loci and mitochondrial (Cyt b and 16S rRNA) genes as a first step towards (i) the detection of potential source populations, and (ii) a better understanding of mechanisms governing evolutionary changes involved in the invasion process. Results based on multivariate analysis (Discriminant Analysis of Principal Components), Bayesian statistical inference (Clustering, Approximate Bayesian Computation) and demographic tests allowed a construction of the introduction pathways of the species over recent centuries. While emigrants originated from only one of the two native lineages, the West one, the most likely scenario involved several introduction events and “source switching” comprising (i) an early stage (around 1660) of simultaneous introductions from Europe (France, Spain) towards Oceania (New Zealand) and California, (ii) from the early 18^th century, a second colonization wave from bridgehead populations successfully established in California, (iii) genetic admixture in invasive areas where highly divergent populations came into contact as in New Zealand. Although these man-made pathways are consistent with historical data, introduction time estimates suggest that the two putative waves of invasion would have occurred long before the first field observations recorded, both in America and in Oceania. A prolonged lag period as the use of an incorrect generation time could explain such 100–150 years discrepancy. Lastly, the contrasting patterns of neutral genetic signal left in invasive populations are discussed in light of possible ways of facing novel environments (standing genetic variation versus new mutation).     

Microgeographical variation in shell size of the land snail Chondrina clienta

Microgeographical variation in shell morphology of the rock-dwelling land snail Chondrina clienta , collected from 30 sites within an area of 0.5 km² on the island of Öland, Sweden, was examined in relation to its own population density and that of a potential competitor (the land snail Balea perversa ) and to environmental factors. Dispersal of marked individuals averaged 83 cm per year within a stone pile and 291 cm in an area of exposed bedrock. Local population density of active C. clienta ranged from 5 to 794 individuals per m². Shell characters were highly intercorrelated, both within and between populations. Principal component analysis revealed that most of the interpopulational variation could be expressed by the single character of shell height, which ranged from 5.54 to 6.94 mm. In all populations, snails of a given size had the same whorl number. Shell size was not influenced by habitat type (exposed rock surface, stone pile or stone wall) or proportion of calcareous stones within habitat. It was, however, negatively correlated with local population density, indicating intraspecific competition, and positively correlated with the degree of plant cover within the habitat. Analysis of variance revealed additionally a density effect of B. perversa on shell size in C. clienta , probably as a result of interspecific competition. Breeding experiments using C. clienta from different sites and carried out under unifrom conditions caused most of the phenotypic variation to disappear, demonstrating the high phenotypic plasticity of the species.     

Elements of cold hardiness in a littoral population of the land snail Helix aspersa (Gastropoda: Pulmonata)

The land snail Helix aspersa can be considered partially tolerant to freezing, in the sense it can survive some ice formation within its body for a limited time, and possesses a limited ability to supercool. This study aimed at understanding what factors are responsible for the variation of the temperature of crystallization ( Tc) in a littoral temperate population. The ability to supercool was maximal (ca. -5 degrees C) during dormancy periods (hibernation and aestivation) and minimal (ca. -3 degrees C) during spring and autumn, in relation with the decrease of water mass and the increase of osmolality. Tc decreased in October to remain stable through late autumn and winter; it increased quickly with the awakening of animals in April. Snails with an epiphragm had a significantly higher ability to supercool (ca. -4.8 degrees C) than snails which did not form an epiphragm (ca. -4.2 degrees C). The animals' size had a weak but significant influence on the realization of the Tc. It appeared that there was not a real cold-hardiness strategy in this population; rather a sum of parameters, varying in consequences of the external conditions and of the activity cycle, which are responsible for the enhancement of the supercooling ability during winter.     

Cold hardiness of some Alpine Collembola

Abstract.

• 1 Individual supercooling points ranged from -2 to -44°C for six species of springtails, five species from the Swiss Alps and one from lowland Britain. Individuals of Isotomurus alticola (Carl) and Isotoma viridis Bourlet without gut contents had substantially lower supercooling points than those containing food material.
• 2 Juveniles were more cold resistant than adults in both I.alticola and Isotoma hiemalis Schött, both with respect to supercooling point and to their survival at prolonged subzero temperatures.
• 3 Temperature and acclimation time affected the degree of supercooling of four of the Alpine species especially I.hiemalis.
• 4 Duration of culture period had no consistent influence on the supercooling potential of all the species.
• 5 Tests for glycerol in the body fluids of the five Alpine springtails were negative, but the presence of a sugar, probably glucose, together with a five carbon polyhydric alcohol was indicated by chromatography.

     

Cold hardiness of the overwintering black carpenter ant

Abstract Cold hardiness was investigated in overwintering field nests of the black carpenter ant Camponotus pennsylvanicus (De Geer) in the Commonwealth of Virginia. No active nest thermoregulation was observed: temperatures of galleries, worker bodies, worker clusters, and larval clusters were within 3°C of ambient temperature. Nest temperatures generally fluctuated less rapidly and severely than did ambient temperature; thus, the nest afforded protection from potentially fatal sudden temperature drops. Glycerol, the only polyol cryoprotectant detected, was found in all castes and larvae. Supercooling points were low and ranged from ? 17°C in major workers to ?22°C in larvae. A second heat release peak, occurring around ? 8°C, was seen in all adults, but it was not observed in larvae. This higher temperature peak in adults probably represents the freezing of the gut contents, as adults were found to overwinter with the crop full or partially full. Larvae did not overwinter with liquid food in the gut.     

Cold hardiness in the scorpion, Paruroctonus aquilonalis.

Lower lethal temperatures represented by supercooling points were measured for the scorpion Paruroctonus aquilonalis (Stahnke) following exposure to laboratory regimes of photoperiod and temperature, as well as to ambient conditions. Analyses for the presence of glycerol and sorbitol in hemolymph, and of body water content were also performed.Laboratory results indicated that neither decreasing photoperiod nor decreasing temperature influenced supercooling point in a meaningful way. However, supercooling points did decrease significantly during the fall and winter among scorpions maintained outdoors, indicating development of cold hardiness. Neither glycerol nor sorbitol were detected, nor were the slight seasonal decreases found in water content considered sufficient to influence supercooling point.Ability to avoid winter freezing in P. aquilonalis appears primarily conferred by substantial seasonal depression of the supercooling point and is temporally associated with cessation of feeding in the fall. Supercooling responses were compared with those of another scorpion, Diplocentrus spitzeri, and differences tentatively were explained in terms of feeding activity.     

Multiple predator-avoidance behaviours of the freshwater snail Physella heterostropha pomila: responses vary with risk

• 1 We examined the predator‐avoidance behaviour, exhibited in response to chemical cues, of two populations of the snail Physella heterostropha pomila. Snails were subjected to four treatments simulating different degrees of predation risk: control water (low risk), or water from tanks containing nonforaging crayfish (intermediate risk), crushed conspecifics (high risk) or crayfish consuming conspecifics (high risk). Data were analysed using three‐way ANOVA models (population × predator chemicals × injured conspecific chemicals).
• 2 Physella increased its avoidance behaviour as risk increased. Crayfish cue elicited a significantly greater response than from controls. Cues from injured conspecifics elicited the strongest response.
• 3 Physella exhibited several types of avoidance behaviour, including burial into the substratum, moving to the water surface, and crawling out of the water. The type of cue present influenced response type. Cues from crayfish reduced burial and increased movement to the water surface or out of the water. Cues from injured‐conspecifics significantly increased crawling completely out of the water.
• 4 The two populations differed in the type and degree of response exhibited. One population exhibited significantly greater ‘reactivity’ (i.e. any avoidance behaviour) in response to foraging crayfish, and more burial and crawl‐out behaviours were exhibited in high‐risk treatments.

     

Cold hardiness and overwintering of the grain aphid Sitobion avenae

ABSTRACT.

• 1 Cold hardiness as measured by supercooling ability in the active stages of the grain aphid Sitobion avenae (F.) decreased progressively with maturation.
• 2 Aphids showed no acclimation response when maintained at low temperatures.
• 3 Starvation did not improve supercooling ability.
• 4 In a single exposure, surface moisture caused inoculation above the inherent supercooling point in a small proportion of a population.
• 5 Field populations show a seasonal change in supercooling ability, which is at a maximum in summer and a minimum in late winter.
• 6 It is concluded that the act of feeding on healthy plant tissue may confer extensive supercooling ability.

     

Cold hardiness of the eggs of the plaice,Pleuronectes platessa

Summary The pelagic eggs of the plaice, Pleuronectes platessa, at the stage of first heart beat, can tolerate freezing into solid sea ice at temperatures down to-6°C. A long term freezing period of 5 days, at sublethal temperatures of-3 to-4°C, had only a minor effect on the eggs. When the ice was absent from the medium, the eggs supercooled readily to temperatures around-15° to-20°C. However, no thermal hysteresis agents were present in the eggs at low temperature and the cold hardiness therefore seem to rely on a high tolerance to brine solutions at low temperatures.     

Self-fertilization in the white-lipped land snail Triodopsis Albolabris

The breeding system of the polygyrid land snail Triodopsis albolabris was studied in laboratory colonies through the use of allozyme-genetic markers. Isolated virgin individuals self-fertilized only after several months of apparent self-sterility. Isolated pairs appeared to reproduce solely by outcrossing. Overall, the normalized reproductive success of paired individuals was about 86 times greater than that of isolates.
Nine natural populations of this snail were surveyed at eight allozyme loci. All populations were highly polymorphic, and heterozygosity was high and agreed with Hardy-Weinberg expectations. This indicates that selfing is not common in established populations. We conclude that T. albolabris avoid inbreeding by selfing if at all possible. However, if the probability of finding a mate is low (assessed by several months without finding one), lone individuals self-fertilize.     

Cold hardiness adaptations of codling moth, cydia pomonella

The cold hardiness adaptations of natural and laboratory reared populations of the codling moth, Cydia pomonella, were examined. Hemolymph, gut, and whole body supercooling points (SCPs), 24-h LT50s, polyhydroxy alcohol concentrations, hemolymph freezing points, and hemolymph melting points were determined. Nondiapausing codling moth larvae do not have appreciable levels of ice nucleators in the hemolymph or gut. Whole body supercooling points were higher than hemolymph supercooling points. For nondiapausing larvae, LT50s were significantly higher than both the whole body and the hemolymph supercooling points, indicating the presence of chill sensitivity. As the larvae left the food source and spun a cocoon, both hemolymph and whole body SCPs decreased. Diapause destined larvae had significantly lower hemolymph SCPs than nondiapausing larvae, but whole body SCPs were not significantly different from nondiapausing larvae of the same age. The LT50s of diapause destined and diapausing larvae were significantly lower than that of nondiapausing larvae. Codling moths are freezing intolerant, with LT50s close to the average whole body supercooling point in diapause destined and diapausing larvae. The overwintering, diapausing larvae effectively supercool to avoid lethal freezing by removal of ice nucleators from the gut and body without appreciable increase of antifreeze agents such as polyols or antifreeze proteins.