Supercooling ability in two populations of the land snail Helix pomatia (Gastropoda: Helicidae) and ice-nucleating activity of gut bacteria

The land snail Helix pomatia (Gastropoda: Helicidae) is widely distributed in Northern and Central Europe where it may experience subzero temperatures during winter months. Its supercooling ability was studied in two populations of H. pomatia. One population originated from Southern Sweden (Gotaland) and the other from Central France (Auvergne). In the experimental design, they were acclimated, over 2 weeks, to artificial winter conditions (hibernation, T=5 degrees C). The Swedish snails showed a rather limited supercooling ability (temperature of crystallization, T(c)=-6.4+/-0.8 degrees C), significantly greater, however, than the supercooling capacity of the population from France (T(c)=-4.6+/-1.4 degrees C). In artificial spring conditions (3 months of hibernation followed by a progressive acclimation, over 2 weeks, to activity at T=20 degrees C), both populations exhibited a similar high T(c) (-2.0+/-1.0 degrees C). The lower T(c) of hibernating Swedish snails could be due to a greater loss of body water, accompanied by a higher concentration of solutes in the hemolymph. In both populations, the variation in hemolymph osmolality measured between hibernating (250-270 mOsm kg(-1)) and active (165-215 mOsm kg(-1)) snails may be explained by the variation in body water mass and did not suggest the production of colligative cryoprotectants. Moreover, the three bacterial strains, Buttiauxella sp., Kluyvera sp., and Tatumella sp. (Enterobacteriaceae) which were isolated from fed snails, but absent in starved snails, did not show any ice-nucleating activity at temperatures higher than -9 degrees C. Only the strain Kluyvera sp. initiated nucleation at -9 degrees C. This strain, therefore, is a weak, also termed a Type III or Class C ice-nucleating active bacterium, but with no influence on the supercooling ability of individual snails. In summary, fluctuations in body water mass of hibernating snail populations, triggering changes in osmolyte concentration, rather than the presence of endogenous ice-nucleating-active bacteria, accounts for fluctuations in their T(c).     

The hemolymph of Biomphalaria snail vectors of schistosomiasis supports a diverse microbiome

The microbiome - the microorganism community that is found on or within an organism's body - is increasingly recognized to shape many aspects of its host biology and is a key determinant of health and disease. Microbiomes modulate the capacity of insect disease vectors (mosquitoes, tsetse flies, sandflies) to transmit parasites and disease. We investigate the diversity and abundance of microorganisms within the hemolymph (i.e. blood) of Biomphalaria snails, the intermediate host for Schistosoma mansoni, using Illumina MiSeq sequencing of the bacterial 16S V4 rDNA. We sampled hemolymph from five snails from six different laboratory populations of B. glabrata and one population of B. alexandrina. We observed 279.84 ± 0.79 amplicon sequence variants per snail. There were significant differences in microbiome composition at the level of individual snails, snail populations and species. Snail microbiomes were dominated by Proteobacteria and Bacteroidetes while water microbiomes from snail tank were dominated by Actinobacteria. We investigated the absolute bacterial load using qPCR: hemolymph samples contained 2784 ± 339 bacteria/μl. We speculate that the microbiome may represent a critical, but unexplored intermediary in the snail–schistosome interaction as hemolymph is in very close contact with the parasite at each step of its development.     

Ecological and biogeographical inferences on two sympatric and enigmatic Andean cat species using genetic identification of faecal samples

The carnivore community of the altiplano ecosystem of the high Andes, including the Andean mountain cat (Leopardus jacobita) and pampas cat (Leopardus colocolo), is one of the least studied in the world. We determined the origin of 186 carnivore samples (184 faeces and two skulls) collected above 3000 m above sea level in northern Chile, including 33 from the Andean mountain cat and 75 from the pampas cat using diagnostic molecular genetic sequence variation. We determined for the first time food habits, habitat and physiographic associations, and general patterns of molecular genetic variation of the Andean mountain cat and the pampas cat in Chile. Both species had narrow dietary niches dominated by small rodents and there was a wide overlap in diet composition (0.82), suggesting low levels of prey partitioning between species. The mountain viscacha (Lagidium viscacia) made up a large proportion of the biomass of the diet of both species, especially for the Andean mountain cat (93.9% vs. 74.8% for the pampas cat), underscoring the importance of further research and conservation focus on this vanishing prey species. Although the probability of finding Andean mountain cat scats increased with altitude and slope, there was substantial geographical overlap in distribution between species, revealing that the pampas cat distribution includes high-altitude grassland habitats. The Andean mountain cat had relatively low levels of mitochondrial DNA (mtDNA) genetic variation (two mtDNA haplotypes) compared with the pampas cat (17 mtDNA haplotypes), suggestive of a distinct evolutionary history and relatively smaller historic populations. These insights will facilitate and provide tools and hypotheses for much-needed research and conservation efforts on these species and this ecosystem.     

Spatial genetics of a high elevation lineage of Rhytididae land snails in New Zealand: the Powelliphanta Kawatiri complex

ABSTRACT

Powelliphanta is a genus of large carnivorous land snails endemic to New Zealand which display phenotypic variation within comparatively small geographic distances. The diversity within these snails has become a matter of high interest to conservation, as many lineages occupy small (or highly fragmented) ranges that render them vulnerable to ongoing habitat loss and predation by exotic pests. Combining Powelliphanta mitochondrial sequence data and genotypes of microsatellite loci we document the genetic structure within a species complex dubbed ‘Kawatiri’. All populations (with one exception) within the Kawatiri lineage are restricted to subalpine habitat (at elevations over 600?m above sea level). The ranges of some Kawatiri complex populations are adjacent to the congeneric lowland species Powelliphanta lignaria. Improved understanding of the distribution of this complex and the level and structure of genetic diversity provided a picture of a naturally fragmented lineage, restricted to a particular ecological zone. We identified six genetic clusters associated with population connectivity orientated north–south along mountain ranges, with east–west divisions between ranges. Future management should aim to retain the evolutionary potential within this young radiation by actively conserving the variation encompassed by each of the six clusters identified here.     

Variation in Terpene and Linear‐Chain Hydrocarbon Content in Yarrow (Achillea millefolium L.) Germplasm from the Rhaetian Alps,Italy

Yarrow (Achillea millefolium L.) is a herbaceous species common in the Alpine region of Europe and used in folk medicine since antiquity. Its organs are rich in monoterpenes and sesquiterpenes, two subclasses of plant terpenoids with relevant ecological significance, which were reported as valuable markers for the traceability of mountain dairy products. The variability in chemical composition of yarrow germplasm may be related with its genetic diversity, accounting for possible differences in medical properties, and supporting its use as a specific territorial marker. Aim of this work was to assess the leaf chemical composition of 16 yarrow populations collected at altitudes exceeding 1600 m in three valleys of the Rhaetian Alps, Italy, and jointly evaluated in a lowland site. The most abundant compounds detected generally differed from those of the germplasm from other countries. A trend of valley‐specific pattern of composition was evident. However, the variability among individual populations was even more remarkable, regardless of their valley of origin. The concentrations of sesquiterpene hydrocarbons, oxygenated monoterpenes, and oxygenated sesquiterpenes discriminated the populations in multivariate analysis. A few prevailing chemotypes were characterized, which differed from those previously reported in the literature. The geographic isolation from other germplasms, and the local ecotypization, likely originated a chemically distinct gene pool.     

Species-richness patterns of vascular plants along seven altitudinal transects in Norway

Altitudinal richness patterns were investigated along altitudinal gradients located in northern Norway (two transects) and along a west–east gradient in southern Norway (five transects). The transects were sampled for vascular plant species richness using a uniform sampling method. Each transect consisted of 38–48 5×5 m sample plots regularly spaced from sea level or valley bottom to a local mountain top. In five transects species richness peaked at mid-altitudes, whereas in the two northern transects species richness decreased with altitude. The observations were qualitatively evaluated in relation to the influence of the area of the species pool, hard boundaries, temperature and precipitation, and mass effect. The observed patterns cannot be fully accounted for by any of these factors. However, the altitude of the peak in species richness was above the forest-limit for all the humped relationships, which may suggest that species richness above the forest-limit might be enhanced by a mass effect from forest taxa. The two monotonic relationships found in the north may be caused by the relatively low number of alpine species at these sites. The monotonic pattern may result from a decrease in "forest species" towards the mountain tops.     

Ecological and life history characteristics predict population genetic divergence of two salmonids in the same landscape

Ecological and life history characteristics such as population size, dispersal pattern, and mating system mediate the influence of genetic drift and gene flow on population subdivision. Bull trout (Salvelinus confluentus) and mountain whitefish (Prosopium williamsoni) differ markedly in spawning location, population size and mating system. Based on these differences, we predicted that bull trout would have reduced genetic variation within and greater differentiation among populations compared with mountain whitefish. To test this hypothesis, we used microsatellite markers to determine patterns of genetic divergence for each species in the Clark Fork River, Montana, USA. As predicted, bull trout had a much greater proportion of genetic variation partitioned among populations than mountain whitefish. Among all sites, FST was seven times greater for bull trout (FST = 0.304 for bull trout, 0.042 for mountain whitefish. After removing genetically differentiated high mountain lake sites for each species FST, was 10 times greater for bull trout (FST = 0.176 for bull trout; FST = 0.018 for mountain whitefish). The same characteristics that affect dispersal patterns in these species also lead to predictions about the amount and scale of adaptive divergence among populations. We provide a theoretical framework that incorporates variation in ecological and life history factors, neutral divergence, and adaptive divergence to interpret how neutral and adaptive divergence might be correlates of ecological and life history factors.     

Total protein and carbohydrate content and protease and disaccharidase activities in the hemolymph of Lymnaea stagnalis naturally infected with digenean larvae

Lymnaea stagnalis is an intermediate host of many Digenea. The infestation affects host metabolism. The aim of the work was to investigate hemolymph biochemical indicators of L. stagnalis infected with four species of trematodes: Diplostomum pseudospathaceum, Paryphostomum radiatum, Plagiorchis elegans or Opisthioglyphe ranae. The protein profiles and proteinase activity in the hemolymph of sexually mature individuals of Lymnaea stagnalis maintained at 19°C were tested. As the carbohydrates are main substrates for energetic metabolism of the great pond snail their content and disaccharidase activity were also studied. Hemolymph samples were collected during weeks 3 and 4 of rearing. No significant differences in the total protein content between uninfected individuals and snails infected with the first three trematode species were detected. In the snails infected with O. ranae the quantity of total proteins was near twice higher than in those uninfected. A higher share of 70 kDa proteins in infected than in uninfected snails as well as reduction of the low molecular weight fractions of proteins for snails infected with D. pseudospathaceum and P. radiatum were detected. During week 3, carbohydrate content in the infected snails did not differ from that in the controls while during week 4 it was significantly lower in snails infected with P. elegans or O. ranae. The content of the major soluble carbohydrate in the hemolymph - saccharose — changed in a similar way. No activity of trypsin or pepsin in the hemolymph sample was detected while the activity of chymotrypsin was lower in infected snails vs. controls. On the other hand, saccharase and maltase activities were higher in infected than in uninfected snails. The biochemical hemolymph indicators in naturally infected host-snails show some differences depending on the parasite species but they are not sufficiently species-specific to offer the basis for establishing the model unique for a particular parasitosis. Our results from the field did not always coincide with those from the laboratory.     

Defence against oxidative stress in two species of land snails (Helix pomatia and Helix aspersa) subjected to estivation

During summer, land snails are exposed to estivation/arousal cycles that imposes oxidative stress, but they exhibit different patterns of antioxidant defence. To test the ability of two related species, Helix pomatia and Helix aspersa, to modulate their antioxidant defence mechanism during estivation/arousal cycles, we examined activities of catalase and glutathione-related enzymes and concentrations of glutathione and thiobarbituric acid reactive substances (TBARS; as products of lipid peroxidation). In both species, estivation evoked changes in activity of total and selenium-dependent glutathione peroxidase (GPx), but did not affect activity of catalase, glutathione reductase, and glutathione transferase, and had no effect on concentration of glutathione. Activity of catalase in estivating snails, instead of the expected increase, showed a tendency to diminish. Extremely low activities of catalase in the foot were usually associated with extremely high activities of both forms of GPx. In conclusion, maintenance of relatively high activities of the antioxidant enzymes and accumulation of glutathione, resulting in a low and stable concentration of TBARS, plays an important role in scavenging oxygen free radicals from the organism of both species.     

雨雪冰冻灾害对南岭蝴蝶资源的影响

南岭国家级自然保护区巳发现蝴蝶442种,其中不乏有中国分布新纪录和新种。常绿阔叶林中有蝴蝶339种,山地常绿针阔叶林中有蝴蝶369种,山顶常绿阔叶苔藓矮林中有蝴蝶71种;这些蝶类属东洋区的有360种,占总数的81.45%;属古北区的有48种,占总数的10.86%;跨区系分布的有34种,占总数的7.69%;南岭蝶类与福建省蝶类相似百分率高达77.27%,说明两地有共同的起源与演化过程。2008年初的雨雪冰冻灾害使南岭海拨500～1000m森林生态遭受严重破坏,各种蝴蝶在林中的种群密度骤降,海拨700～800m的蝶类仅为海拨1100～1200m冻害较轻林地蝶类数量的23%;整个林区蝶类的种群密度极低,与常年相比,降低率约为90%。凤蝶科的种群在调查地段发现的只占常年见到的37.5%。     

o-Diphenol oxidase activity of molluscan hemocyanins

Diphenoloxidase activities of two molluscan hemocyanins, isolated from the marine snails Rapana venosa and garden snails Helix vulgaris were studied using o-diphenol and L-Dopa as substrates. The dimers of H. vulgaris Hc show both, diphenol (K(m)=2.86 mM and K(cat)=4.48) and L-Dopa activity due to a more open active sites of the enzyme and better access of the substrates. The K(m) value of molluscan H. vulgaris Hc is very close to those of Helix pomatia and Sepia officinalis Hcs, but several times higher compared to those of Rapana and Octopus Hcs. Also HvH has a very high enzyme activity compared with other molluscan Hcs. Kinetic measurements with native RvH and both structural subunits, RvH1 and RvH2, show that RvH and only one structural subunit, RvH2, exhibited only o-diphenol activity, but no L-Dopa oxidizing activity.     

Schistosoma mansoni: Lysozyme activity in Biomphalaria glabrata during infection with two strains

Levels of lysozyme activity were determined in the hemolymph, digestive gland, and headfoot extracts of M-line stock of snails, Biomphalaria glabrata, during infection with the PR-1 and Lc-1 strains of the trematode, Schistosoma mansoni. At 3 hr postexposure there was a 10-fold increase in the levels of enzyme activity in the hemolymph of snails infected with the Lc-1 strain to which the snail is resistant. This increase was considerably higher when compared to the threefold increase in the PR-1-infected snails. The infection also induced a gradual depletion of lysozyme activity in the headfoot muscles of the two groups of infected snails. There were no changes in the levels of enzyme activity in the digestive gland extracts of the control and the two groups of infected snails. Similar changes in the levels of enzyme activity in the hemolymph and headfoot extracts of infected snails suggest a nonspecific response to a parasite infection and do not indicate that lysozyme is primarily responsible for the destruction of schistosome parasite in a resistant snail host.     

Parasitic mites influence intra‐ and interpopulational variation in sperm length in a simultaneous hermaphrodite land snail (Gastropoda: Helicidae)

Sperm morphology can be highly variable among species, but less is known about patterns of population differentiation within species. Sperm morphology is under strong sexual selection, may evolve rapidly, and often co‐varies with other reproductive traits that differ between populations. We investigated variation in sperm morphology in the simultaneous hermaphrodite land snail Arianta arbustorum in relation to parasitic mite infection. Variation in total sperm length and sperm head length was assessed in 23 populations sampled across the distributional range of the species in Central and Northern Europe. We found a pronounced variation in total sperm length among the populations studied, with a difference of 11.0% of total sperm length between the shortest and longest population means. Differences among populations explained 62.9% of the variance in total sperm length, differences among individual snails within population 23.4% and differences within individual snail 13.7%. Mantel tests showed that interpopulation differences in total sperm length increased significantly with geographical distance between populations. A minimal adequate model revealed that parasitic infection had a positive effect and longitude a negative effect on total sperm length. Thus, independent of the population examined, mite‐infected individuals of A. arbustorum produced larger sperm than uninfected snails and total sperm length decreased from west to east. Sperm head length also varied among populations, but it was not influenced by any of the factors examined. In a subsample of 12 populations restricted to the mountains of Switzerland (elevational range 440–2485 m a.s.l.), total sperm length decreased with increasing elevation. Our results suggest that selection pressures acting among populations may differ from those acting within. Stabilizing selection might be a possible mechanism for producing the reduced variation observed in sperm length within a population. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 1036–1046.     

The second population of Arctic charr Salvelinus alpinus complex (Salmoniformes, Salmonidae) in the Lake Baikal basin, the highest mountain charr population in Russia

We report the finding of the second population of Arctic charr in Lake Baikal basin, in a nameless lake in the Barguzin mountain range in the outlet of the Svetlaya River. The lake is situated at an altitude of 1766 m above sea level; this is the highest lake inhabited by this species in the territory of Russia. Two abundant charr forms, dwarf and small, were recorded in the lake; data on their ecology, morphology, age composition, growth, and microsatellite variation are presented. Both forms feed mainly on zooplankton, though neither has pronounced morphological traits characteristic of specialized plankton-feeders. Dwarf and small charrs are rather close in meristic characters and in the length of gill rakers, but differ substantially in allele frequencies and allele diversity at the microsatellite loci, which is indicative of a high degree of reproductive isolation between them. The discovered population is the only one among Transbaikalian charr populations, that, due to its remoteness, has not been affected by man. For that reason, it can be considered as an etalon of an undisturbed natural charr population. The lake and its surroundings should receive official protected status.     

海南岛热带山地雨林种类组成的局域分布与垂直分布

对海南岛四大山地雨林区常见植物种类的局域及垂直分布进行了研究,结果表明,海南岛热带山地雨林的种类组成,除了在各林区有一定数量相同的种外,各林区之间种类成分也存在着显著的差异,由于各林区生态因子的差异,使得生态幅度不同的物种在垂直方向上的分布也产生明显的差异,生态幅度较小的物种仅局限于某一地段,生态幅度较广,适应性较强的种类则有较广泛的分布。     

Life history adaptation of Tanymastix stagnalis (Crustacea,Branchiopoda) to habitat characteristics

Life histories of two populations of Tanymastix stagnalisfrom sites in central Italy, differing in climate and altitude above sea level, were compared to obtain information on the tolerance limits of this species.Temperature was the main factor affecting the biology of Tanymastix stagnalis. Significant differences in growth patterns occurred at different sites. A colder climate induced delayed hatching, slower differentiation and maturation, but a longer life span in the mountain population (Forca Canapine) than in the plain (Fosso dei Mergani).Both populations exhibited an initial fluctuating sex ratio, which later became female biased, a pattern which could be the consequence of adaptation to the unstable nature of the biotopes studied.     

Impact of geographical origin on mating behaviour in two species of Biomphalaria (Planorbidae: Gastropoda)

Studies of inbreeding and outcrossing have traditionally concentrated on matings within populations. The influence of geographical origin on mate choice in animals from different populations has received less attention. We investigated whether planorbid snails mated preferentially within their own population or with snails from other populations. Snails from three Biomphalaria pfeifferi strains and three B. glabrata strains were allowed to mate with conspecifics in the laboratory. We recorded their matings at night using time-lapse video. When they could choose between sympatric and allopatric snails, Biomphalaria snails significantly preferred the former: snails of each population mated more often with sympatric than with allopatric snails. This tendency to avoid outcrossing may indicate that, in some species, local adaptations can be more valuable than genetic novelties. Copyright 1999 The Association for the Study of Animal Behaviour.     

西藏特有灌木砂生槐繁殖生长对海拔和沙埋的响应

砂生槐是西藏特有植物,主要分布在雅鲁藏布江中游地区及拉萨河,年楚河等主要支流宽谷区,为评价高原海拔引起的水热条件变化对砂生槐繁殖和生长的影响,沿雅鲁藏布江河谷600多公里海拔从3139m到3925m的样带上,对砂生板天然种群进行了调查,结果表明：沿着高原河谷海拔升市制 梯度,降水量由600mm下降到400mm,年平均气温由8.2℃下降到5.8℃,砂生槐的种群密度趋与增加,其径趋于减小,但植株高度和新枝长度并未随海拔升高呈明显升降变化。种子库密度,种子千粒重,实生苗密度,萌蘖苗密度也没有随海拔高度的变化而呈明显的升降变化。砂生板适应海拔引起的水热条件的变化在生殖方面并为作出大的调整,但在适应风沙干扰方面表现了繁殖方式的调整,即在未受沙埋时,种子繁殖趋于占主导地位,在生境受到沙埋时,营养繁殖趋于占主导地位,繁殖方式的调整可能是砂生槐适应风沙干扰的最主要对策。     

中国四川二郎山地区有瓣蝇类地理垂直分布

1980~2000年调查四川二郎山地区有瓣蝇类(Calyptratae)地理垂直分布。共获成蝇7科88属264种,本文优选其中代表性蝇类30种,以分析其地理垂直分布情况。结果发现因各蝇种特有生态习性的不同而各自在不同垂直地带表现不同的虫口数量分布。     

Isolation and characterization of cultivable fermentative bacteria from the intestine of two edible snails, Helixpomatia and Cornu aspersum (Gastropoda: Pulmonata)

The intestinal microbiota of the edible snails Cornu aspersum fSyn: H. aspersa), and Helix pomatia were investigated by culture-based methods, 16S rRNA sequence analyses and phenotypic characterisations. The study was carried out on aestivating snails and two populations of H. pomatia were considered. The cultivable bacteria dominated in the distal part of the intestine, with up to 5.10(9) CFU g -1, but the Swedish H. pomatia appeared significantly less colonised, suggesting a higher sensitivity of its microbiota to climatic change. All the strains, but one, shared >/= 97% sequence identity with reference strains. They were arranged into two taxa: the Gamma Proteobacteria with Buttiauxella, Citrobacter, Enterobacter, Kluyvera, Obesumbacterium, Raoultella and the Firmicutes with Enterococcus, Lactococcus, and Clostridium. According to the literature, these genera are mostly assigned to enteric environments or to phyllosphere, data in favour of culturing snails in contact with soil and plants. None of the strains were able to digest filter paper, Avicel cellulose or carboxymethyl cellulose (CMC). Acetogens and methanogenic archaea were not cultivated, so the fate of hydrogen remains questionable. This microbiota could play important roles in the digestive process (fermentation) and the energy supply of the snail (L-lactate, acetate). The choice of cereals and plants by snail farmers should take into account the fermentative abilities of the intestinal microbiota.