Morphometric and genetic analysis as proof of the existence of two sturgeon species in the Guadalquivir river

Morphometric and genetic methods were used to identify two sturgeon species, Acipenser naccarii Bonaparte, 1836, and A. sturio Linnaeus, 1758, captured in some of the principal rivers of the Iberian Peninsula, including the Guadalquivir. After measuring 25 Iberian specimens from a fishery and several Spanish and Portuguese museums and applying stepwise discriminant analysis (SDA), four specimens preserved in different museums [two specimens from the Guadalquivir river (EBD-8173 and EBD-8174), one specimen from the Tagus river (MUC1) and one specimen from the Mondego river (MUC46B)], as well as five specimens captured in the Guadalquivir river in the 1940s but not preserved (CM1, CM2, CM3, CM4 and CM5), were identified as A. naccarii. After cloning and characterisation of a satellite-DNA family, HindIII, from A.␣naccarii genome, its absence from the genome of A.␣sturio was determined. Using this satellite-DNA as a genetic marker and by means of dot-blotting, we demonstrate that the DNA of the two specimens captured during the mid-1970s in the Guadalquivir river cross-hybridised with HindIII satellite-DNA sequences of A.␣naccarii. We conclude that A. naccarii is autochthonous to the Iberian Peninsula and is not, as was previously believed, endemic to the Adriatic Sea. Received: 28 November 1996 / Accepted: 10 March 1997     

Sardine (Sardina pilchardus) spawning seasonality in European waters of the northeast Atlantic

Egg data from ichthyoplankton monitoring sites in the western English Channel (1988–2003) and northern Spain (1990–2000) and macroscopic maturity data from biological samples of purse seine landings in western and southern Iberia (1980–2004) are used to describe the spawning seasonality of sardine (Sardina pilchardus) in European waters of the northeast Atlantic using generalised additive models. The fitted models reveal a double peak in spawning activity during early summer and autumn in the western Channel, a wider spring peak off northern Spain and a broad winter season in the western and southern Iberian Peninsula. At all sites, a high probability of spawning activity was observed over at least 3 months of the year, with the duration of the season increasing with both decreasing latitude and increasing fish size. Off western and southern Iberia there are indications that the spawning season has been of longer duration in recent years for all size classes (reaching in some cases 8 months of the year for large fish). These patterns are in general agreement with existing literature and theoretical expectations of sardine spawning being driven locally by the seasonal cycle of water temperature, assuming preferences for spawning at 14 –15°C and avoidance for temperatures below 12°C and above 16°C. Regional quotient plots indicated that spawning tolerance to higher temperatures increases progressively with decreasing latitude. Despite the weak evidence for geographical differences in temperature tolerance that may have some genetic origin, the degree of spatio-temporal overlap in sardine-spawning activity within Atlantic European waters is unlikely to promote any reproductive isolation in that area.     

Ultrastructure of the spermatozoa of Hymenosomatidae (Crustacea: Brachyura) and the relationships of the family

The spermatozoa of the genus Odiomaris Ng and Richer de Forges, 1996 (=Amarinus Lucas, 1980) have the components typical of eubrachyuran (Heterotremata + Thoracotremata) sperm, but differ significantly from all other investigated eubrachyurans in at least ten characteristics: (1) presence of an epiopercular dome; (2) separation of all but the central region of the operculum from the remainder of the acrosome by an infra-opercular rim; (3) the fact that the acrosome is smaller in volume than the nucleus; (4) the acrosome is strongly emergent from the nucleus, being surrounded only basally by nuclear material; (5) the cytoplasmic sheath, ending anteriorly with the nucleus, is also basal; (6) division of the acrosome contents into an inner and outer acrosome zone is scarcely apparent in longitudinal section as the inner zone is narrow and of doubtful homology; (7) the thin, putative inner acrosome zone is anteriorly almost septate owing to several longitudinal corrugations; (8) basally there is a unique “fringe zone”; (9) the acrosome, including the epinuclear dome, is longer than wide; (10) the unique helical and posterolateral disposition of the nuclear arms. From a purely spermatological viewpoint, Odiomaris (as exemplified by O. pilosus and O. estuarius), and provisionally the Hymenosomatidae, are thus excluded from the Thora- cotremata, in which they were formerly placed, nor are they readily placeable in the Heterotremata. Received: 30 December 1996 / Accepted: 4 February 1997     

GC-EAG-analysis of volatiles from Brussels sprouts plants damaged by two species of Pieris caterpillars: olfactory receptive range of a specialist and a generalist parasitoid wasp species

Summary. Feeding by Pieris brassicae or P. rapae caterpillars on Brussels sprouts plants induces the emission of synomones that attract natural enemies of the caterpillars, Cotesia glomerata, a generalist parasitoid, and C. rubecula, a specialist on P. rapae. Previous research on this tritrophic system has identified a large number of volatiles in the headspace of herbivore-damaged Brussels sprouts plants, and this paper addresses the question which of these volatiles are perceived by the two parasitoid species. Headspace odors from both P. brassicae- and P. rapae-damaged Brussels sprouts plants were analyzed by coupled gas chromatography electro- antennogram (GC-EAG) detection. Twenty volatiles evoked consistent EAG reactions in the antennae of both species and nineteen of these volatiles could be identified with GC-MS. One component that could not be identified due to its low concentration, evoked EAG responses in antennae of C. rubecula only. Possible consequences for searching behavior of the two parasitoid species are discussed.     

马氏甲烷八叠球菌S－6的一个染色体区段的表达、序列分析和结构分析

用一组多克隆抗体对马氏甲烷八叠球菌（Ｍｅｔｈａｎｏｓａｒｃｉｎａｍａｚｅｉ）Ｓ－６菌株的基因组ＤＮＡ文库进行了筛选．仅选出ｐ６０Ａ克隆能对马氏甲烷八叠球菌中可发生细胞形态学变化菌株的抗血清发生阳性反应．对ｐ６０Ａ克隆的双链ＤＮＡ进行了序列分析．识别出一开式阅读框架（ｏｐｅｎｒｅａｄｉｎｇｆｒａｍｅＰ，ＯＲＦＰ）．表达的蛋白是ＯＲＦＰ编码的蛋白的３倍，并得到ＯＲＦＰ蛋白的三聚体，在ＳＤＳ－ＰＡＧＥ中表现出整体蛋白的迁移行为．同时，此表达蛋白抗１０％ＳＤＳ，６ｍｏｌ／Ｌ尿素及热处理．基因结构分析表明，ＯＲＦＰ具有与Ｍ．ｂａｒｋｒｉｍｃｒＡ有同源性的核糖体结合位点和一个与甲烷细菌启动子共有序列相似度达７７％的启动子序列．使用参照序列分析表明，由ＯＲＦＰ演绎的氨基酸序列，具有高密度的带电荷的氨基酸和占优势的β－层迭构型．对此表达蛋白作了Ｎｅｕｒｏｓｒｏｒａｃｒａｓｓａ的ｐｏｒｉｎ蛋白抗血清试验，以研究其可能功能．检验了Ｍ．ｍａｚｅｉＳ－６细胞的蔗糖梯度制备物，对此原细胞中的ＯＲＦＰ蛋白作了定位．结果表明，ＯＲＦＰ蛋白可能是一种古细菌Ｐｏｒｉｎ，其在Ｍ．ｍａｚｅｉＳ－６中的表达，可能象大肠杆菌那样与渗透压调节有关．     

Bacterial colonization and endocytosis on the gill of a new limpet species from a hydrothermal vent

Calcification rates in different fragments along branches of the hermatypic coral Stylophora pistillata were tested in the laboratory using a new technique, the optic glassfiber method. By this method, the tested colony remains constantly in dark conditions while a narrow beam of light, transferred through the optic fiber, illuminates a small distinct point of coral tissue (on a branch tip or base). The selected illuminated portion of the branch serves as the experimental fragment, while all the other parts of the same colony serve as the dark controls. The results indicate that significantly more calcium is incorporated in the tip fragments than in the bases, both in light and in dark conditions (4.1 to 13.2 times more). Illumination of the tips or the bases did not stimulate or enhance the calcification rates of these fragments. Thus, in all colonies tested, the calcification rates of the illuminated fragments were not significantly different from the average rates of other similar, non-illuminated fragments of the same colony. It is suggested that light does not directly enhance calcification in hermatypic corals, but rather, that light enhances O₂ production, which consequently stimulates coral metabolism. Our preliminary results indicate that calcification rates recorded in aerated dark experiments are significantly higher than calcification rates of non-aerated dark controls.     

Double-labelling methods used to diagnose apoptotic and necrotic cell degradations in copepod nauplii

Two double-labelling methods, Tunel+propidium iodide and Annexin V-FITC+propidium iodide, have been tested to diagnose cell degradation processes in N1–N2 nauplius stages produced by spawning females of the copepod Calanus helgolandicus fed either non-toxic (PRO: control) or toxic (TR1) diets under laboratory conditions. Observation of labelled samples with a confocal laser-scanning microscope revealed that the maternal-food effect, following absorption of the toxic diet, induced cell apoptosis and necrosis in 80–100% of the offspring, which sooner or later died. Similar cell damages were also observed in 18–50% of the nauplii produced by females incubated in filtered seawater. Such cell degradation processes and high mortality were not observed in nauplii produced by females fed the non-toxic diet. Our protocols allowed detection of cell degradations before death of unhealthy larvae, preferentially with a confocal microscope or a conventional visible-fluorescent-light microscope.Communicated by O. Kinne, Oldendorf/Luhe