Sexual reproduction in the compound ascidian Diplosoma listerianum (Tunicata). II. Sperm penetration through ovary wall and evidence of internal fertilization

Diplosoma listerianum differs from most ascidians in that, at ovulation, eggs are emitted at the bottom of the ovary and segregated into the tunic, so that fertilization occurs far from seawater. A fertilization canal, a hollow extension of the ovary, conducts sperm towards the egg. In the present paper, ultrastructural evidence is reported on the morphological relationship between the ovary, egg envelopes and oocyte and on the mechanism by which sperm-egg interaction is established. In the ovary, the very complex sperm, equipped with a spiral dense groove, undergo metamorphosis as the first step in a sperm reaction and then pass through the ovary epithelium insinuating themselves between the intercellular junctions which appear to be mouldable, although able to maintain the egg-ovary barrier. Sperm then reach the vitelline coat, where a further step in the sperm reaction occurs. Before the egg abandons the ovary, the sperm head is incorporated into the oocyte by a process recalling phagocytosis, with the formation of an engulfing pocket. Sperm-egg contact and incorporation in D. listerianum occur in a way, never previously reported for other ascidians, in which fusion of plasma membranes takes place immediately after sperm-egg contact. Unlike other cytoplasmic components, the dense groove persists until the sperm enters the egg. It gives a corkscrew-like configuration to the sperm head and allows close adhesion to cell membranes, facilitating sperm movement. Expulsion of numerous cortical granules and features of a cortical reaction were observed in the egg penetrated by the sperm. The mode of internal fertilization of this species in comparison with that of other tunicates and phylogenetic aspects are discussed. Ripe colonies of D. listerianum collected in the Lagoon of Venice, Italy in 1986 and 1991 and colonies reared on glass in aquaria were used for our investigations.     

Environmental Accumulation of Airborne Fluorides in Romania

The nature and extent of pollution from an aluminium smelter and a fertiliser factory in Romania were studied. These are large industrial complexes, and both types of industry are known to release fluorides into the atmosphere. In grass samples collected from around the aluminium smelter, the maximum fluoride levels were found to be 4023mgkg^–1 and 162mgkg^–1 in unwashed and washed grass samples respectively, and 89mgkg^–1 in soils. For the fertiliser factory, the maximum levels in washed grasses were found to be 207mgkg^–1, and 11mgkg^–1 in the soils. In both locations, these maximum values were obtained in samples collected from within 200m of the factory limits, and compare with regional background levels of less than 10mgkg^–1 for grasses and 2mgkg^–1 for soils. The high fluoride levels of fluoride in the grasses are sufficient to give cause for concern for the effects that these could have on the local population and on grazing animals.     

Spermatozoon structure of some North American prosobranchs from the families Lottiidae (Patellogastropoda) and Fissurellidae (Archaeogastropoda)

The spermatozoa of four species of the patellogastropod family Lottiidae (Lottia pelta, L. digitalis, L. strigatella, Tectura scutum) and one species of the archaeogastropod family Fissurellidae (Diodora aspera) were examined in 1990 using transmission electron microscopy. All have primitive or ect-aquasperm, typical of invertebrates using external fertilization. Sperm of the lottiid limpets are characterized by a 5 to 9 m-long head composed of a conical acrosome which constitutes >50% of the head length, and a cylindrical nucleus. The acrosome of all species of lottiids is differentiated internally, and has a posterior invagination 0.9 to 1 m in depth, into which an elongate acrosomal lobe protrudes. Between the posterior acrosomal lobe and the nucleus, the subacrosomal material is aggregated as a fibrous column. The midpiece of the sperm has a ring of 4 to 5 spherical mitochondria of 0.6 m diam, posterior to which is a collar of cytoplasm 1 m long, which sheaths the anterior portion of the axoneme. The size and morphology of the acrosome and large cytoplasmic collar clearly distinguish the spermatozoa of the Lottiidae from other families of Patellogastropoda. The sperm of D. aspera (Fissurellidae) is typical of the family of archaeogastropod; the head has a length to breadth ratio of 4:1, and the cylindrical nucleus is capped by a small acrosome, <25% of the total head length, which is deeply invaginated.     

Temporal variation and distribution of trace metals in freshwater and fish from Calabar River, S.E. Nigeria

An investigation on the abundance and distribution of trace metals (Fe, Cu, Zn, Mn, Cr, Cd and Pb) in water, and nine species of fish samples from Calabar river was carried out in 1992. The concentrations of iron (6000–7240gl^–1), zinc (4910–7230gl^–1), and cadmium (3–7gl^–1) showed moderate pollution while those of copper (420–630gl^–1), manganese (23–48gl^–1), chromium (<10–20gl^–1) and lead (<1–10gl^–1) in water were well below WHO permissible levels. Significant seasonal changes (0.001p0.25) were obtained for iron, copper, zinc, manganese and cadmium in water. Furthermore, iron, zinc and cadmium showed statistically significant spatial changes (0.005p0.10). Of the nine fish species studied, no statistically significant relationship between body weight and the concentrations of the metals was observed. The concentrations of the metals per mean total body weight apparently decreases in the order Fe>Zn>Cu>Mn>Pb>Cd=Cr and were within the limits that were safe for consumption.     

Analysing sperm competition data: simple models for predicting mechanisms

Summary Prospective models are developed for analysing sperm competition data so as to predict the underlying mechanisms determining paternity in multiply mated females. The models require: 1) estimations of proportion of offspring sired by the last male to mate (P ₂), 2) knowledge of the number of sperm transferred by each male, and 3) knowledge of the sperm storage capacity of the female, should this be limited. They will distinguish between raffles (sperm mixing without displacement) and sperm displacement mechanisms. The sensitivity of the techniques can be increased by manipulating the number of sperm transferred by each male. Typically, this can be done by manipulating copula duration or number of ejaculations, given a knowledge of the rate of sperm transfer. Data from two contrasting insect species are fitted to the models to demonstrate the techniques. These models are prospective only, and their limitations are discussed. The principal limitation is that we assume that sperm used for fertilization mix randomly in a fertilization set immediately prior to fertilization; in reality this may be difficult to identify. When sperm mixing is very rapid, the fertilization set will often be equivalent to the sperm stores, but with slow mixing, the fertilization set may be equivalent to a much more restricted zone and may change with time. Offprint requests to: G.A. Parker     

Male capacity as related to sperm production,pregnancy initiation,and sperm competition in deer mice (Peromyscus maniculatus)

Summary Limitations on male capacity may be important to the evolution of mating strategies and behavior. Sperm counts in successive ejaculates of deer mice (Peromyscus maniculatus) decreased progressively through six ejaculations and remained depressed the following day. Males were as effective in inseminating females in cycling estrus with their second two ejaculates as with their first two and as effective in inseminating postpartum females with their fifth as with their first. In sperm competition situations males competed with rested males as effectively with their fourth, fifth, and sixth ejaculates as with their first. Thus, although sperm counts decrease in successive ejaculates, males suffer no apparent functional deficiency as long as they continue ejaculating. It is not clear whether or not deer mice are honest salesmen but they appear to be honest ejaculators and usually are honest intromitters.     

Primary production in Dumbell Bay in the Arctic Ocean

Photosynthesis, chlorophyll, inorganic nutrients, and related factors were measured throughout a productive season in a small coastal inlet of the Arctic Ocean. Significant production was confined to 5 m for a very limited time period beacause of both light and nutrient limitations. Maximum chlorophyll was 8.2 mg Chl a m^-3 and maximum gross and net photosynthesis rates were 830 and 550 mg C m^-2 d^-1. Annual gross and net photosynthesis is estimated at about 12 and 9 g C m^-2 respectively. The effect of light stimulation on assimilation is described, sources of coastal zone nutrient enrichment are considered, and discrepancies between gross and net photosynthesis are discussed with reference to the growth characteristics of the phytoplankton population.Bigelow Laboratory for Ocean Sciences. Contribution No. 76008     

Aqua regia extractable trace elements in surface soils of venezuela

Surface soils (0–15 cm) were sampled at 10–20 km intervals along two transects in Venezuela. One (1162 km, 70 samples) ran west to east parallel with the Caribbean coastline, the other (920 km, 92 samples) ran south to north from the frontier with Brazil to the Caribbean shore. Sampling took place in both a wet and a dry season. Trace metals were extracted from dried, sieved (<2 mm) soil with boiling aqua regia followed by analysis by ICP or flame AAS. Metal values did not differ significantly between the two seasons and dates were averaged. Geometric mean values for the west–east transect were: Cr=41.5, Cu 17.9, Cs 3.6, Li=13.9, Mn=294, Ni=21.3, Pb=17.4, Sr=39.4, V=60.4 and Zn = 83.7g g^–1, respectively. Similarly, for the south–north transect Cr=21.3, Cu=4.3, Cs=1.1, Li=2.0, Mn=55.7, Ni=4.4, Pb=6.1, Sr=13.3, V=28.2 and Zn=16.7g g^–1, respectively. A classification of samples by lithology showed surface soil composition to be related to rock composition. Metal values were low in the soils in the south of the country, in the Guyana highlands (Gran Sabana). Low Zn contents were prevalent. Lead contents were affected by roadside fallout from vehicles using leaded petrol except that high Pb contents of soils in the Gran Sabana were of more complex origin.     

Evaluation of Public Housing Lead Risk Assessment Data

A unique data set from lead risk assessments performed on 67 public housing developments from across the United States was made available for analyzes. The data set includes results of lead analysis from 5906 dust wipes and from 1222 soil samples. A total of 487 dwelling units in these developments, as well as associated common areas, were sampled, all by the same team of inspectors. The number of dwelling units within a development that were sampled reflected the guidelines then in force, the 1990 Interim HUD Guidelines, rather than those specified in the 1995 Guidelines. Median dust lead loadings for floors, 151gm^–2 (14gft^–2), and window sills, 936gm^–2 (87gft^–2), were much less than former HUD limits of 1076gm^–2 (100gft^–2) and 5380gm^–2 (500gft^–2), respectively and are only about one-third of the recently established limits of 431gm^–2 (40gft^–2) and 2690gm^–2 (250gft^–2). In contrast, the median lead loading for window troughs, 8560gm^–2 (795gft^–2), was almost identical to the HUD clearance limit of 8610gm^–2 (800gft^–2). There was a strong positive correlation between floor and window trough lead loading values for samples from the same dwelling units and those from common areas of the housing developments. Door threshold samples, which may reflect conditions exterior to the dwelling unit, were collected from 53 dwelling units. Median lead loading levels of these samples were more than ten times higher than those in floor samples from the same dwelling units, were about the same as window sill samples and about one-half of levels in window trough samples. Composite sample results, simulated by averaging results from four samples within a dwelling unit, revealed that in order to have the same rate of excedence of standards, the composite standards would have to be reduced, for example, from the single sample value of 1076gm^–2 (100gft^–2) to 527gm^–2 (49gft^–2) for floor samples and from the single sample value of 8610gm^–2 (800gft^–2) to 5160gm^–2 (479gft^–2) for window troughs. For this public housing data set, the portion of the units in developments containing more than 225 units which exceeded the established limit for window samples was the same when using either the full data set or a random one-half of the data set. This suggests that, for this data set, the number of dwelling units sampled was excessive . Thus, the required increase in the number of dwelling units to be sampled specified in the 1995 Guidelines for developments with more than 225 dwelling units, may not have been necessary if this data set is representative of public housing developments in the United States.     

Paternity assurance by “helpful” males: adaptations to sperm competition in burying beetles

Summary Male burying beetles invest parentally by participating in the burial of a carcass and in provisioning and guarding the larvae that come to the carcass. Since most of the females arriving at a carcass have stored fertile sperm within their spermathecae, sperm transferred by such helpful males must compete with other males' ejaculates for the fertilization of the female's eggs. We showed that these males are able to achieve a high level of paternity (mean=92%). The mechanism they employ is a repeat-mating tactic, i.e., the female is mated very frequently shortly before and during oviposition. Repeated matings are essential for a high reliability of paternity, since single copulations result in the fertilization of only a very small proportion of the female's eggs.     

Why do male Callosobruchus maculatus beetles inseminate so many sperm?

Male Callosobruchus maculatus F. (Coleoptera: Bruchidae) inseminate more sperm than females can effectively store in their spermathecae. This study examines the adaptive significance of excess sperm transfer by measuring components of male and female reproductive success in response to manipulating the number of sperm inseminated. The number of sperm transferred during copulation was reduced from 56,000 ±4,462 to 8,700±1,194 by sequentially mating males to virgin females. Reducing the number of sperm inseminated by the first male to mate had no effect on the extent of sperm precedence, but reducing the number of sperm inseminated by the second male resulted in a significant reduction in the extent of sperm precedence. When large numbers of sperm are inseminated the remating refractory period of females is increased. These results indicate that males transferring large numbers of sperm during copulation have a two-fold advantage at fertilization; they are more effective at preempting previously stored sperm and they are likely to father more offspring by delaying the time of female remating. The transfer of excess sperm does not appear to serve as nonpromiscuous male mating effort; the number of eggs laid, their fertility and the subsequent survival of zygotes were unaffected by manipulating the number of sperm inseminated. The underlying mechanisms of sperm precedence were also examined. Simple models of sperm displacement failed to accurately predict the patterns of sperm precedence observed in this species. However, the results do not provide conclusive evidence against the models but rather serve to highlight our limited understanding of the movement of sperm within the female's reproductive tract.     

Indoor and outdoor household dust contamination in Cincinnati, Ohio, USA

Despite some progress in reducing the average lead level in the USA, the streets of Cincinnati, Ohio, are still contaminated by heavy metals. High levels of heavy metals will have significant unequivocal ecological impacts and pose a potential health hazard. This study evaluates the level of heavy- metal contamination in household dust and examines its relationships with the external environment. Samples of outdoor and indoor dust were collected from middle-income residential homes in the Greater Cincinnati Metropolitan District and the metal content was analysed by inductively coupled plasma atomic emission spectrophotometry. Results showed that the mean levels of lead in outdoor and indoor dusts are 650 g g-1 and 377 g g-1 and the copper levels are 253 g g-1 and 510 g g-1, respectively. The median levels are 156 g g-1 and 139 g g-1 for outdoor and indoor lead dusts and 35 g g-1 and 124 g g-1 for outdoor and indoor copper dusts. The degree of contamination may be ascribable to the age of the dwelling unit and the neighbourhood, the time when the unit was last painted, the presence of pets and the type of space heating. In addition, car exhausts seems to be a possible source of contamination.     

The distribution of heavy metals in deposited urban dusts and sediments, Coventry, England

Samples of urban dusts and sediments have been collected and categorised into three compartment those which act mainly as sources, those undergoing transport and those which have been deposited. The sediments were fractionated into <63m to match similar sized lacustrine sediments and <2mm to represent natural and anthropogenic fluvially transported sediments. Preliminary results of total heavy metals analysis show that the <2mm fraction of the source group exhibits the greatest variabilty in metal concentrations. Overall, however, the <63m fraction dominates regardless of compartment. Some heavy metal concentrations decrease through the sourcetransportdeposit process, possibly due to selective onward transport of finer material or other anthropogenic processes unique to the urban environment (e.g. street cleansing). Sitespecificity is indicated in a comparison with other similar studies.     

Environmental Geochemistry of Soils and Waters of Susaki Area, Korinthos, Greece

Soil and water samples were collected from the Susaki area of Korinthos and analysed for heavy metals in order to evaluate their environmental impact. The geology of the studied area includes ultrabasic rocks and Neogene and Quaternary deposits whereas magnesite veins are found within the ultrabasic rocks. In the north part of the studied area post volcanic emissions of H₂S, CO₂ and H₂O vapor continue to the present day. All the samples were analysed for heavy metals by the ICP method. The element ranges (in g g^–1) for soil samples are: Cu 11–63, Pb 5–256, Zn 21–604, Ni 183–2665, Co 12–124, Mn 456–1434, As 5–104, Sr 44–730, V 21–84, Cr 163–2346, Ba 48–218, Zr 3–41, Y 3–13. The metals Pb, Zn, Ni, Co, Cr, Fe, Cu, Mn, As and Sr are enriched in the Susaki soils. The element ranges for water samples are: Cu 65–103ppb, Pb<10ppb, Zn<5ppb, Ni 21–163ppb, Co 2–12ppb, As<30ppb, Cr<20ppb, Ba 36–785ppb, Sb<10ppb, W<10ppb, Bi<30ppb, Mn 0.0–0.9 g g^–1, Fe 0.01–0.22 g g^–1, Na 843–3076 g g^–1, K 98–278 g g^–1, Si 39–65 g g^–1, P 0.1–0.2 g g^–1. There is a natural pollution of soils with elevated concentrations of Ni, Co, Mn, Fe and Cr due to the presence of ultrabasic rocks. Another natural case of As pollution of soils is due to the volcanic activity and the geothermal field in the area. The geochemical data of ground waters and also the ¹⁸, D data showed a mixing in different proportions between sea water and meteoric water.     

Lead Contamination of Topsoil and Vegetation in the Vicinity of a Battery Factory in Nigeria

Levels of Pb, Zn, Cu, Cr, Ni and Cd were determined in topsoil and vegetation in the vicinity of a factory manufacturing lead- batteries in Ibadan, Nigeria. The samples were collected along five transects in different directions, and varying distances up to 1000m from the factory. Soil lead levels were found to be elevated around the factory, with average levels of about 2000mgkg^–1 close to the fence that declined gradually to about 50mgkg^–1 some 750m away. Soil-lead level around a primary school located about 500m from the factory was as high as 1450mgkg^–1. Lead levels were equally elevated in the vegetation, though average levels in vegetation were slightly lower than in the soil. Cadmium concentrations in soil and vegetation, though low, were more positively correlated with lead levels than any of the other metals are with lead.     

“Copper” granules in the barnacle Balanus balanoides

Balanus balanoides (L.) collected from an area with high heavy-metal run-off contained two different types of granule within the parenchyma cells of the prosoma. X-ray microprobe analysis shows one to be the familiar zinc granule made up of concentric layers and giving major peaks for phosphorus and zinc, and the other to be homogeneous and giving peaks for sulphur and copper. This latter granule is designated the copper granule. Whilst zinc granules are known to be composed of inorganic phosphate, various tests on copper granules in sections and in a granule-rich pellet have shown that the copper is probably complexed with organic matter. The tests also demonstrated the relatively, insoluble (inert) nature of these granules. Although zinc and copper granules were present together in the prosoma, atomic absorption analyses of whole bodies (prosoma+thorax) have shown the level of zinc (50.28 g/mg dry weight) to be much higher than that of copper (3.75 g/mg dry weight).     

Populational distribution I: The digital simulation

This paper presents a method of investigating the distributional pattern of a biological population, using a technique of simulation. The method consists of the comparison of the empirical frequency curve of a population, obtained using the Method of Quadrats, with a simulated one, since the pattern of the simulated curve depends on the simulated distribution. An example is given.     

Copper effects on reproductive stages of Baltic Sea Fucus vesiculosus

Copper is an active ingredient in many antifouling products, and pleasure boats are estimated to be the major single source of copper pollution in Swedish coastal waters. For this reason, the effects of copper were studied on egg volume, fertilization, germination and development of apical hairs of Baltic Sea Fucus vesiculosus L. Germination was the most sensitive stage and was studied at different concentrations of copper, different salinities and different ages of zygotes. Low concentrations of copper, 2.5 g Cu l^-1, added to natural brackish water before fertilization, adversely affected germination at the ambient, suboptimal salinity of 6, suggesting that as little as a doubling of the copper levels in the studied area will severely affect the germination frequency of F. vesiculosus. The addition of 20 g Cu l^-1 caused about 70 to 80% decline in germination at 6S but also at 20S which is higher than optimum. At a salinity close to optimum (14S) no negative effect was noticed on germination when 20 g Cu l^-1 was added. The results suggest that the degree of salinity stress acting upon the zygotes is a more important factor for the response to copper than the influence of salinity on metal availability. When 2.5 to 60 g Cu l^-1 was added to the medium 24 h after fertilization, the zygotes were more resistant, resembling the response of adult marine fucoid tissue.     

Arsenic tissue concentration of immature mice one hour after oral exposure to gold mine tailings

A potentially high bioavailability of arsenic in gold mine tailings from a site in northern California has been suggested by solubility studies. To help address this issue, an in vivo dosing study was conducted using 12dayold Swiss Webster mouse pups (n=8/group). A sample of sizefractionated mine tailings from the site (<20m particle size, 691g g^–1 arsenic) was prepared as an aqueous suspension and administered by gavage in a volume that provided 4mg As/kg body weight. The control group received the same volume of a commercial soil (1g g^–1 As) of similar particle size (<60m). No mortality or toxic signs were noted in either group. Tissue samples were collected 1h after gavage, freezedried, microwavedigested and analysed for arsenic by ICP/MS (detection limit 2ng As g^–1 dry weight). Arsenic concentrations (ng As g^–1 dry weight) in tissues from the pups who received mine tailings were significantly higher than in control tissues. The mean elevation in arsenic concentration was highest in the liver (3364% of control, p<0.0001), followed by blood (818 of control, p<0.0001), skin (207% of control, p=0.07), and brain (143% of control, p<0.0001). The carcass arsenic concentration (excluding the GI tract, liver, brain and skin) was 138 of control (p=0.02). The data indicate uptake of arsenic from weathered mine tailings by the immature mouse pups after oral exposure.     

Age-structure dependence of the “specific production” in zooplankton populations

This account represents the results of investigations on the production of some zooplankton populations. For comparing different populations the index specific production (the production rate of the unit of the biomass) is used. The correlation between specific production and age-structure for some populations is described. On the basis of the growth theory of Von Bertalanffy (1938) a theoretical model which helps to explain the relation between specific production and age-structure of the population is proposed.