Studies on a PMCA-like protein in the outer mantle epithelium of <Emphasis Type="Italic">Anodonta cygnea</Emphasis>: insights on calcium transcellular dynamics

Early studies on the outer mantle epithelium (OME) cells of the freshwater bivalve Anodonta cygnea (Linnaeus, 1758) revealed high ionic calcium concentrations by electrophysiological methods and subsequently a high tendency to reach an intracellular toxic condition. This toxicity could be neutralized by specific mechanisms in the cytosol of OME cells of A. cygnea. The present immunocytochemistry studies of OME cells by light and transmission electron microscopy (TEM) clearly showed a positive reaction of an antibody directed against the human plasma membrane Ca²⁺-ATPase 1 (PMCA-1) in the cytoplasm of OME cells. Also, western blot analysis of different fractions of OME cells with anti human PMCA-1 and C28R2 antibodies confirmed the presence of a PMCA-like protein with an unusual topographical localization and a molecular weight of only 70–80 kDa. These results lead us to speculate that this PMCA-like protein is distributed either in the plasma membrane or in the entire cytosol, where it eventually regulates intracellular calcium levels. Interestingly, the antibody reactions showed seasonal variations, being highest in OME samples prepared during summer when A. cygnea live under natural acidosis and absent in samples taken in winter conditions, which is in accordance with the seasonal variation of shell calcification rates. During winter, PMCA-1 antibody reaction was also detected in OME cells of animals kept only under experimentally induced acidosis conditions. Therefore, we assume that a functional role for this PMCA-like protein in the intracellular calcium regulation of OME cells during the mineralization of the shells of A. cygnea can be speculated.     

Phylogeography of the genus <Emphasis Type="Italic">Ulva</Emphasis> (Ulvophyceae,Chlorophyta), with special reference to the Japanese freshwater and brackish taxa

The nuclear-encoded ITS and associated 5.8S rDNA regions were sequenced for 72 specimens of Ulva collected from 44 rivers across Japan, including U. prolifera Müller from the Shimanto River, Kochi Prefecture, as well as 26 samples originally identified as U. linza L. from 20 coastal marine areas. Sequence data revealed that the samples fall into six distinct clades: the U. flexuosa Wulfen clade (2 samples), the Ulva linza-procera-prolifera (LPP) complex clade (75 samples), Ulva sp. 1 clade (3 samples), Ulva sp. 2 clade (7 samples), Ulva sp. 3 clade (4 samples) and Ulva sp. 4 clade (7 samples). The LPP complex contained a mixture of 26 samples collected from seashores and 49 samples obtained from rivers, including U. prolifera from the Shimanto River, and GenBank data for U. linza and U. procera Ahlner. The samples of the LPP complex differed by only 0–7 substitutions (0–1.149%). Subsequent phylogeographic analyses of the LPP complex based on the 5S rDNA spacer region revealed the presence of two further groupings: a group including 22 strictly marine littoral U. linza samples and a U. prolifera group composed of a mixture of 4 marine samples and all 49 river samples. The monophyly of all river samples indicates that adaptation to low salinity might have occurred only once in the evolutionary history of the LPP complex.     

Influence of soft rush (Juncus effusus) on phosphorus flux in grazed seasonal wetlands

Livestock significantly affect wetland soils and vegetation but their impacts on wetland nutrient dynamics are poorly understood. We set up a full factorial laboratory experiment to assess the effects of Juncus effusus, grazing exclusion, and flooding on P flux from intact cores collected from seasonal wetlands in cattle pastures in south Florida. We collected intact cores from Juncus tussocks and plant interspaces inside and outside 4-year grazing exclosures in five replicate wetlands. We incubated the cores for 50 days under continuous flooding or weekly 1-day flooding cycles and measured P concentrations in surface and pore water. Grazing exclosures had less Juncus (17%) and bare ground (2%) than adjacent grazed areas (Juncus, 48%; bare ground, 12%), but did not affect P fluxes. Initial fluxes of soluble reactive P (SRP) were much higher in cores with Juncus (242 ± 153 mg P m⁻² day⁻¹) than without Juncus (14 ± 20 mg P m⁻² day⁻¹). In weekly flooded cores P fluxes fell to 19.7 ± 13.4 mg P m⁻² day⁻¹ in cores with and 2.7 ± 2.6 in cores without Juncus. The strong effect of Juncus on P flux was an indirect effect of cattle grazing, but 4 years of grazing exclusion did not have a significant effect on P fluxes.     

Volume and morphology changes of a bdelloid rotifer species (Macrotrachela quadricornifera) during anhydrobiosis

Following a study on the changes occurring in a bdelloid species (Macrotrachela quadricornifera, Rotifera, Bdelloidea) when entering anhydrobiosis, we investigated the changes in morphology, including weight and volume during the transition from the active hydrated to the dormant anhydrobiotic state by scanning electron microscopy, confocal microscopy and light microscopy. We compared sizes and morphologies of hydrated extended, hydrated contracted and anhydrobiotic specimens. Bdelloid musculature is defined: longitudinal muscles are contracted in the hydrated contracted animal (head and foot are retracted inside the trunk), but appear loose in the anhydrobiotic animal. When anhydrobiotic, M. quadricornifera appears much smaller in size, with a volume reduction of about 60% of the hydrated volume, and its internal organization undergoes remarkable modifications. Internal body cavities, clearly distinguishable in the hydrated extended and contracted specimens, are no longer visible in the anhydrobiotic specimen. Concomitantly, M. quadricornifera loses more than 95% of its weight when anhydrobiotic; this is more than expected from the volume reduction data and could indicate the presence of space-filling molecular species in the dehydrated animal. We estimate that the majority of body mass loss and volume reduction can be ascribed to the water loss from the body cavity during desiccation.     

The approach to equilibrium of multilocus genotype diversity under clonal selection and cyclical parthenogenesis

Sexual generations in cyclical parthenogens are typically separated by multiple generations of clonal reproduction. In contrast to sexual reproduction, during parthenogenesis the genome of the parent is passed on to the offspring as a unit. The absence of recombination during parthenogenesis leads to differences in the action of natural selection in the two reproductive phases. In addition, since recombination is a sampling process, random genetic drift is potentially more important in sexual reproduction than in parthenogenesis. A recent development in the study of rotifer population genetics is the use of microsatellites to characterize natural populations. Microsatellites are selectively neutral, show patterns of Mendelian inheritance and tend to be much more variable than allozymes. An advantage over allozymes is that microsatellite DNA can be cloned with PCR and thus multiple loci can be assayed from a single individual. We use a new computer model in this paper to investigate the response of selectively active and selectively neutral genes to evolutionary forces during cyclical parthenogenesis. Selectively active alleles may respond differently to selection in the parthenogenetic and sexual phases of cyclical parthenogenesis. Even when strong clonal selection is acting on loci associated with adaptation, the view that emerges with microsatellites may be one of Hardy-Weinberg and linkage equilibrium. Thus studies using selectively neutral loci may fail to detect clonal selection even when it is an important feature of the rotifer population's adaptive structure.     

Recipes for successful anhydrobiosis in bdelloid rotifers

We tested the effect of several environmental variables on the ability of three bdelloid rotifers (Macrotrachela quadricornifera, Philodina roseola and Adineta oculata) to recover from the anhydrobiotic state. The variables we examined were (1) rate of water evaporation, (2) relative humidity during anhydrobiosis, (3) temperature during anhydrobiosis, (4) duration of anhydrobiosis, and (5) rehydration rate. Our results indicate that bdelloids can regulate to some degree net water balance during onset and termination of anhydrobiosis.     

Lake characteristics influence recovery of microplankton in arctic LTER lakes following experimental fertilization

Lakes N-1 and N-2 at the Arctic Long Term Ecological Research site at Toolik Lake, Alaska, U.S.A. were fertilized with nitrogen and phosphorus for 5 and 6 years, respectively. The response and recovery of the microplankton community (protozoans, rotifers and crustacean nauplii) differed in the two lakes. Microplankton biomass in Lake N-1 increased five-fold while that in Lake-N-2 only doubled, despite larger nutrient additions to N-2. Microplankton community structure in Lake N-1 shifted toward dominance by few taxa, while the community in Lake N-2 maintained diversity. Finally, the recovery of Lake N-1 to near prefertilization microplankton biomass levels was rapid, while Lake N-2 showed at least a 1-year lag in recovery. These differences appear to be related to differences in the structure of lake sediments.     

Egg size and offspring fitness in a bdelloid rotifer

To test if the quality of offspring is affected by egg size and whether it increases with the amount of resources allocated to an individual egg, we compared the offspring hatched from small and large eggs of Macrotrachela quadricornifera(Rotifera, Bdelloidea). Differently sized eggs were obtained by feeding mothers with different food concentrations. Large eggs were expected to provide better offspring in terms of (1) development time of the embryos, (2) age at first reproduction of the juveniles and (3) recovery after desiccation of the newborns. The comparison between offspring hatched from large vs. small eggs of the same bdelloid species revealed that animals hatched from large eggs had shorter embryonic development and earlier age at first reproduction than animals hatched from small eggs. In contrast, the capacity to survive stress, like desiccation, was not affected by egg size. Thus, offspring hatched from larger eggs had shorter generation times, but were not favoured under harsh circumstances.     

利用RAPD标记研究几种淡水腹足类的亲缘关系

以软体动物腹足纲、中腹足目的光滑狭口螺、长角涵螺、纹沼螺、赤豆螺、大沼螺几个形态分类的近缘种及梨形环棱螺(对照)作为研究对象,用随机扩增多态DNA技术对上述动物进行了20个引物的扩增,共获得117条扩增谱带,单个引物扩增的RAPD标记在3-12个之间,片段长度在30—3000bp之间。试验结果显示：淡水螺类的RAPD标记具有明显的多态性,而且种间的扩增标记及差异程度可以反映出物种间系统演化过程的亲缘关系。通过数值聚类制图后得到：长角涵螺、纹沼螺、大沼螺、赤豆螺之间的亲缘关系较近,而光滑狭口螺、梨形环棱螺与上述4个种的亲缘关系较远,其结果与新修订的淡水中腹足目科级分类方案相当吻合。     

海口地区集贸市场淡水鱼华枝睾吸虫囊蚴感染调查

用直接压片法和人工消化法检查了海口集贸市场上5种淡水色(鲫色、罗非鱼、鲤鱼、土鲮色、白鲳)感染华枝睾吸虫囊蚴情况。结果表明：5种淡水色的总感染率为53．66％。其中,鲤鱼、土鲮色、鲫鱼、罗非鱼、白鲳的感染率分别为68．75％、58．82％、58．06％、48．57％和36．67％;平均每克鱼肉含囊蚴数最高的是鲫鱼(9．47个／g),平均每尾阳性鱼含囊蚴数最高的是土鲮鱼(43个／尾)。