Marine bacterial fouling: A scanning electron microscope study

A range of substrates has been examined in order to determine the types of attachment mechanism employed by marine bacteria. Use of the scanning electron microscope (SEM) has also allowed an investigation of the initimate relationship between an antifouling paint matrix and its attached bacterial community. Plastic (Thermanox) and glass coverslips, together with Cu₂O-based and TPTF-based antifouling paints and their respective empty-matrix analogues have been used in this study. Observations over periods of up to 4 wk have shown that extensive bacterial communities can develop. A variety of bacteria have been found: cocci; various rods; stalked forms; and prosthecate types. These bacteria also exhibit a range of attachment mechanisms. Initial attachment is by simple stickiness of cell walls, flagella, pili (fimbriae) or stalks. This stickiness can be attributed to an actual adhesive, electrostatic forces, electrical double-layer phenomena or to London/van der Waals forces. Often, attachment is subsequently improved by the secretion of insoluble, high molecular weight, polysaccharidic material. This material is found in the form of strands, pads, sheets or capsules and acts to bridge the space between the cell wall and substrate or adjacent cells. Thus, stalked forms are found attached by basal pads of mucilage whilst cocci and rods are often found enmeshed in mucilage strands and sheets, but less often attached by pads and capsules. Initially, single bacterial cells attach and give rise to colonies by cell division. Continuing growth of adjacent colonies leads to the development of confluent films over the substrate surface. Further growth results in thickening until eventually the entire surface is coated with a dense community of mixed bacteria together with their adhesive mucilage. In the case of antifouling paints, the porous nature of the matrix allows invasion by bacteria and the possibility of pore blockage by adhesive polysaccharides. This problem is discussed with reference to the paint's loss of antifouling efficiency.     

New epizooic symbioses between sponges of the genera Plakortis and Xestospongia in cryptic habitats of the Caribbean

Three new cases of sponge symbiosis between species of Plakortis and Xestospongia were found in reef caves and mesophotic reef habitats of the Caribbean. Plakortis sp. 1 from the Bahamas associates exclusively with Xestospongia deweerdtae which was originally described living freely on the deep fore-reef and caves of Jamaica. In addition, we found Plakortis sp. 2 from Puerto Rico which associates with both X. deweerdtae and a different Xestospongia sp. Sponge specimens were identified using cytochrome oxidase subunit 1, 28S rRNA and 18S rRNA gene sequence fragments, spicule analysis, and histological sections with SEM. Unlike previous sponge pairs, Xestospongia spp. not only grew as a thin veneer of tissue over the Plakortis host sponge but through the mesohyl, forming inner channels (0.1–1 cm) that may provide a benefit by facilitating more efficient water transport through the dense Plakortis tissue. Symbioses with both Plakortis spp. were documented from an early recruit stage through adulthood. Spicule measurements conducted on symbiotic versus free-living X. deweerdtae revealed significantly smaller spicule sizes for symbiotic individuals, suggesting a cost in terms of silicon availability, or a benefit in terms of a lower investment in skeleton synthesis for support. This study reveals new specialized symbiotic associations between distantly related sponge genera that likely represent an alternative strategy of adaptation for life in reef caves and mesophotic reefs.     

Settlement of barnacle larvae: surface structure of the antennular attachment disc by scanning electron microscopy

The surface morphology of the antennular attachment disc of the cypris larva of Balanus balanoides has been investigated by scanning electron microscopy, and its function is considered in the context of settlent behaviour. It is concluded that the attachment organ acts as an adhesive pad for temporary attachment while the larva explores the substratum. The surface of the disc is covered with villi, which could retain an adhesive substance thought to be produced by the antennular glands. The open-ended hair of the axial sense organ on the disc is shown.     

Invasive Species Research to Meet the Needs of Resource Management and Planning

Abstract: As zebra mussels (Dreissena polymorpha) continue to spread among inland lakes of the United States and Canada, there is growing interest from professionals, citizens, and other stakeholders to know which lakes are likely to be colonized by zebra mussels. Thus, we developed a classification of lake suitability for zebra mussels on the basis of measured or estimated concentrations of dissolved calcium in lake water and applied the classification to >11,500 lakes in Wisconsin and the Upper Peninsula of Michigan. The majority of lakes (58%) were classified as unsuitable (<10 mg/L Ca) for survival and reproduction of zebra mussels, 27% were identified as suitable (≥21 mg/L Ca), and 15% were classified as borderline suitable (≥10 and <21 mg/L Ca). Of the 77 inland lakes with confirmed zebra mussel records for which data on dissolved calcium were available, our method classified 74 as suitable and 3 as borderline suitable. To communicate this lake‐specific suitability information and to help prioritize regional efforts to monitor and prevent the expansion of zebra mussels and other invasive species, we developed a web‐based interface (available from http://www.aissmartprevention.wisc.edu/ ). Although we are still uncertain of how access to suitability information ultimately affects decision making, we believe this is a useful case study of building communication channels among researchers, practitioners, and the public.     

An antifouling catechol/chitosan-modified polyvinylidene fluoride membrane for sustainable oil-in-water emulsions separation

• Underwater superoleophobic membrane was fabricated by deposition of catechol/chitosan. • The membrane had ultrahigh pure water flux and was stable under harsh pH conditions. • The membrane exhibited remarkable antifouling property in O/W emulsion separation. • The hydration layer on the membrane surface prevented oil droplets adhesion. Low-pressure membrane filtrations are considered as effective technologies for sustainable oil/water separation. However, conventional membranes usually suffer from severe pore clogging and surface fouling, and thus, novel membranes with superior wettability and antifouling features are urgently required. Herein, we report a facile green approach for the development of an underwater superoleophobic microfiltration membrane via one-step oxidant-induced ultrafast co-deposition of naturally available catechol/chitosan on a porous polyvinylidene fluoride (PVDF) substrate. Membrane morphology and surface chemistry were studied using a series of characterization techniques. The as-prepared membrane retained the original pore structure due to the ultrathin and uniform catechol/chitosan coating. It exhibited ultrahigh pure water permeability and robust chemical stability under harsh pH conditions. Moreover, the catechol/chitosan hydrophilic coating on the membrane surface acting as an energetic barrier for oil droplets could minimize oil adhesion on the surface, which endowed the membrane with remarkable antifouling property and reusability in a cyclic oil-in-water (O/W) emulsion separation. The modified membrane exhibited a competitive flux of ~428 L/(m²·h·bar) after three filtration cycles, which was 70% higher than that of the pristine PVDF membrane. These results suggest that the novel underwater superoleophobic membrane can potentially be used for sustainable O/W emulsions separation, and the proposed green facile modification approach can also be applied to other water-remediation materials considering its low cost and simplicity.     

Microstructures of Antarctic cidaroid spines: diversity of shapes and ectosymbiont attachments

The echinoderm endoskeleton, located in the connective layer of the tegument, is organized into a three-dimensional mesh, the stereom. Among echinoids, the cidaroids depart from this pattern, and the shaft of the spine lacks an epidermis. Thus, the spines lack antifouling protection, allowing ectosymbionts such as bryozoans and foraminiferans to attach. This raises a question about the adaptive role of the cortical layer of the stereom. This study examined the micro- and mesostructure of the spines of 11 cidaroid species collected in the Weddell Sea and Drake Passage, and the nature of their ectosymbiont attachments. Scanning electron microscopy was used to characterize the cortex surface and X-ray micro computed tomography (μCT) to describe the symbiont attachments. Spine microstructure features provide a useful taxonomic character for distinguishing among three species in the genus Ctenocidaris, and challenge a previous parasitic interpretation of cortical filaments on the spines of Rhynchocidaris triplopora. Ectosymbiont attachments were classified as Anchoring, Molding, Cementing, or Corroding. The study suggests that some microstructure features may be protective, keeping the ectosymbionts away from the cortex and loosely attached at intervals along the shaft of the spine, while other micro-structures facilitate attachment over considerable areas of the shaft. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Autotoxic effects of essential oils on photosynthesis in parsley, parsnip, and rough lemon

Summary. Many plant species contain essential oils with allelochemical properties, yet the extent to which these same chemicals can be autotoxic is unclear. In this study, we tested the toxicity of several essential oil components to three species that produce them—Pastinaca sativa and Petroselinum crispum (Apiaceae), and Citrus jambhiri (Rutaceae). The effects of exogenous application of small amounts of essential oil components to the surface of foliage, followed by a pinprick to allow entry into the leaf, were monitored by chlorophyll fluorescence imaging. Rapid and spatially extensive declines in photosynthetic capacity were detected within 200 s. The most toxic constituents were monoterpenes. Two sesquiterpenes, caryophyllene and farnesene, and the phenylpropanoid myristicin, by comparison, were not toxic. Autotoxicity of endogenous essential oil was investigated by slicing through containment structures (glands or tubes); significant toxicity, manifested by reduced photosynthetic activity, was observed in all three species but was most pronounced in P. sativa and P. crispum, both of which possess oil tubes.     

Inhibitory effects of red algal extracts on larval settlement of the barnacle Balanus improvisus

We examined the chemical antifouling properties of four sublittoral red algae, Chondrus crispus, Delesseria sanguinea, Osmundea ramosissima, and Polyides rotundus, which are all rarely fouled in the field. Two different approaches were used. Firstly, we tested the effects of lipophilic crude extracts on the settlement behaviour of cyprid larvae of the co-existing barnacle Balanus improvisus. Secondly, in a settlement preference experiment, we tested whether B. improvisus cyprid larvae settle on living algae when given a choice between natural algal surfaces and control surfaces. With this procedure, we were able to test both if the algae inhibit recruitment of cyprids, and if this inhibition is a result of chemistry. The settlement of B. improvisus larvae was strongly inhibited at concentrations estimated to be potentially ecologically relevant for all of the tested extracts. However, only C. crispus significantly inhibited settlement in the preference experiment, even though there was also a tendency for settlement inhibition on P. rotundus and O. ramosissima. In contrast, D. sanguinea seemed to stimulate settlement. This contradiction probably resulted from an extraction of metabolites that naturally occur only inside the alga. However, as this study shows, a combination of settlement assays with whole-cell extracts and preference tests of ecologically relevant fouling organisms on natural algal and control surfaces may be a useful procedure to avoid erroneous conclusions regarding natural antifouling roles of compounds based on settlement assays with only whole-cell extracts. Furthermore, this study also shows that production of inhibitory metabolites may explain the low degree of fouling, especially by B. improvisus, on C. crispus.Communicated by L. Hagerman, Helsingør     

Hydrodynamic stress and habitat partitioning between indigenous (Perna perna) and invasive (Mytilus galloprovincialis) mussels: constraints of an evolutionary strategy

The ability of a mussel to withstand wave-generated hydrodynamic stress depends mainly on its byssal attachment strength. This study investigated causes and consequences of different attachment strengths of the two dominant mussels species on the South African south coast, the invasive Mytilus galloprovincialis and the indigenous Perna perna, which dominate the upper and the lower areas of the lower balanoid zone, respectively and co-exist in the middle area. Attachment strength of P. perna was significantly higher than that of M. galloprovincialis. Likewise solitary mussels were more strongly attached than mussels living within mussel beds (bed mussels), and in both cases this can be explained by more and thicker byssal threads. Having a wider shell, M. galloprovincialis is also subjected to higher hydrodynamic loads than P. perna. Attachment strength of both species increased from higher to lower shore, in response to a gradient of stronger wave action. The morphological features of the invasive species and its higher mortality rates during winter storms help to explain the exclusion of M. galloprovincialis from the low shore. The results are discussed in the context of the evolutionary strategy of the alien mussel, which directs most of its energy to fast growth and high reproductive output, apparently at the cost of reduced attachment strength. This raises the prediction that its invasive impact will be more pronounced at sites subject to strong but not extreme wave action.     

Invasive species impacts on ecosystem structure and function: A comparison of Oneida Lake, New York, USA, before and after zebra mussel invasion

Exotic species invasion is widely considered to affect ecosystem structure and function. Yet, few contemporary approaches can assess the effects of exotic species invasion at such an inclusive level. Our research presents one of the first attempts to examine the effects of an exotic species at the ecosystem level in a quantifiable manner. We used ecological network analysis (ENA) and a social network analysis (SNA) method called cohesion analysis to examine the effect of zebra mussel (Dreissena polymorpha) invasion on the Oneida Lake, New York, USA, food web. We used ENA to quantify ecosystem function through an analysis of food web carbon transfer that explicitly incorporated flow over all food web paths (direct and indirect). The cohesion analysis assessed ecosystem structure through an organization of food web members into subgroups of strongly interacting predators and prey. Our analysis detected effects of zebra mussel invasion throughout the entire Oneida Lake food web, including changes in trophic flow efficiency (i.e., carbon flow among trophic levels) and alterations of food web organization (i.e., paths of carbon flow) and ecosystem activity (i.e., total carbon flow). ENA indicated that zebra mussels altered food web function by shunting carbon from pelagic to benthic pathways, increasing dissipative flow loss, and decreasing ecosystem activity. SNA revealed the strength of zebra mussel perturbation as evidenced by a reorganization of food web subgroup structure, with a decrease in importance of pelagic pathways, a concomitant rise of benthic pathways, and a reorganization of interactions between top predator fish. Together, these analyses allowed for a holistic understanding of the effects of zebra mussel invasion on the Oneida Lake food web.     

Marine succession of fouling organisms in Hong Kong,with a comparison of woody substrates and common,locally-available,antifouling paints

Successional stages of macrofaunal fouling organisms of test blocks were exposed in the subtropical waters of Hong Kong. A comparison of 5 types of wood commonly used to build boats, and 5 types of locally available antifouling paints is made. Rafts holding the test blocks were exposed at 1 m depth from 24 March, 1970, to 23 October, 1970. The initial macrofoulers on the control (chinese fir) were Balanus and Bugula, and the final dominance was a Styela-Mytilus community. On painted surfaces, the Balanus-Elasmopus community was dominant. Temperature, salinity, dissolved oxygen, and turbidity were recorded weekly during the test. A preliminary chemical analysis of percentage pigment and cuprous oxide content in the antifouling paints is presented.     

Sulfonated graphene nanomaterials for membrane antifouling,pollutant removal,and production of chemicals from biomass: a review

Water pollution and the unsustainable use of fossil fuel derivatives require advanced catalytic methods to clean waters and to produce fine chemicals from modern biomass. Classical homogeneous catalysts such as sulfuric, phosphoric, and hydrochloric acid are highly corrosive and non-recyclable, whereas heterogeneous catalysts appear promising for lignocellulosic waste depolymerization, pollutant degradation, and membrane antifouling. Here, we review the use of sulfonated graphene and sulfonated graphene oxide nanomaterials for improving membranes, pollutant adsorption and degradation, depolymerization of lignocellulosic waste, liquefaction of biomass, and production of fine chemicals. We also discuss the economy of oil production from biomass. Sulfonated graphene and sulfonated graphene oxide display an unusual large theoretical specific surface area of 2630 m²/g, allowing the reactants to easily enter the internal surface of graphene nanosheets and to reach active acid sites. Sulfonated graphene oxide is hydrophobic and has hydrophilic groups, such as hydroxyl, carboxyl, and epoxy, thus creating cavities on the graphene nanosheet’s surface. The adsorption capacity approached 2.3–2.4 mmol per gram for naphthalene and 1-naphthol. Concerning membranes, we observe an improvement of hydrophilicity, salt rejection, water flux, antifouling properties, and pollutant removal. The nanomaterials can be reused several times without losing catalytic activity due to the high stability originating from the stable carbon–sulfur bond between graphene and the sulfonic group.

     

Evaluation of developmental toxicity and apoptotic induction of the aqueous extract of Millettia pachycarpa using zebrafish as model organism

Extensive and indiscriminate use of synthetic compounds and natural compounds obtained from plant sources have resulted in serious threats to the aquatic ecosystem and human health. Aqueous extract of the root of the plant, Milletia pachycarpa Benth, is currently used for killing fish in the state of Manipur, India. Moreover, this plant is also used as traditional medicine in this region. Although it is widely used in traditional medicine, there is limited information available regarding the adverse effects and mechanism underlying its toxicity. This study examined the effects of exposure to aqueous extract of M. pachycarpa (AEMP) on early embryonic development of zebrafish embryos and mechanisms underlying toxicity. Zebrafish embryos treated with different concentrations of the AEMP produced embryonic lethality and developmental defects. The 96-hr-LC₅₀ of AEMP was found to be 4.276 µg/mL. Further, multiple developmental abnormalities such as pericardial edema, yolk sac edema, spinal curvature, swim bladder deflation, decreased heart rate, and delayed hatching were also observed in a dose-dependent manner. Zebrafish embryo showing moderate-to-severe developmental defects following AEMP exposure cannot swim properly. Further, this study examined oxidative stress and apoptosis in embryos exposed to AEMP. Enhanced production of ROS and apoptosis was found in brain, trunk, and tail of zebrafish embryos treated with AEMP. Data suggest that oxidative stress and apoptosis are associated with AEMP-induced embryonic lethality and developmental toxicity in zebrafish embryos.     

Monitoring the effectiveness of the European tributyltin regulation on the Basque coast (northern Spain) by assessing imposex in two gastropod species (Nassarius reticulatus and Nassarius nitidus)

Imposex and butyltin body burden were assessed in 2011 along the Basque coast (northern Spain) in two gastropod species (Nassarius reticulatus and Nassarius nitidus) four years after an initial survey in 2007. The aim of this re-survey was to monitor the effectiveness of the European ban on the use of tributyltin (TBT) based antifouling paints on ships’ hulls (EC 782/2003). Imposex levels in 2011 were lower than those determined in 2007 at most of the sampling sites. Accordingly, TBT concentrations in the female body burden of Nassarius reticulatus varied from 43 to 250 ng Sn/g in dry weight in 2011, which was a lower maximum than in 2007. Nevertheless, the results for the butyltin degradation index suggest that there have been recent inputs of TBT within the two main Basque harbours. Overall, the legislative measure is contributing to the reduction of TBT effects on the Basque coast although its presence is still evident.     

Strengthening effects of ammonia nitrogen on the harmless biological treatment of oily sludge

To improve the efficiency of oil degradation and strengthen the harmless treatment of oily sludge, three dominant strains identified as Chryseomicrobium sp. YL2, Gordonia sp. YL3 and Acinetobacter sp. YL5 were isolated from soil near a refinery, and the effects on the bioremediation of the oily sludge from the refinery were investigated. The results showed that the efficiency of oil degradation increased by 31.5% compared with the control when the dominant strains were added to the treatment of oily sludge. Furthermore, the dominant strains could use oil as a carbon source for heterotrophic nitrification–aerobic denitrification. The addition of ammonia nitrogen resulted in a large number of remaining microbes and heightened dehydrogenase activity in the oily sludge, further accelerating oil degradation, mainly for C₁₁ to C₂₅ saturated hydrocarbons, and the oil degradation efficiency increased by 40.8%. After 120 days of bioremediation, the biotoxicity of oily sludge, which was expressed by the equivalent phenol concentration, decreased by 40.0% compared with that of the control, indicating that the addition of ammonia nitrogen enhanced the biodegradation of oil. This method can be used to strengthen the harmless treatment of oily sludge in practical engineering applications.     

Assessing stress response of Stylophora pistillata towards oil and phosphate pollution in the Gulf of Aqaba,using molecular and biochemical markers

Crude oil (from oil terminal) and raw phosphate (from phosphate port) pollution are responsible for the lowered health conditions of coral reefs at their vicinity in the Jordanian coast of the Gulf of Aqaba. Both in situ incubations and ex situ laboratory exposure experiments were used to study the effects of those pollutants on corals, by using molecular and biochemical biomarkers in the coral Stylophora pistillata. For ex situ part of the experiment, crude oil and raw phosphate were added to a final concentration of 500?ppm for both pollutants. The DNA damage was assessed by Comet assay, while biochemical stress markers were reassessed by lipid peroxidation (LPO) test. Although the corals looked healthy from outside, the use of stress biomarkers indicated that they are under high pressure at the cellular level. The corals incubated with oil and phosphate had more DNA damage and LPO in comparison with the control samples. The results obtained suggest that the use of stress biomarkers can be used as important prognostic tools for examining the sub-lethal stress on corals before their death.     

Effects of nano carbon black and single-layer graphene oxide on settlement,survival and swimming behaviour of Amphibalanus amphitrite larvae

The effects of two carbon-based nanomaterials, nano-sized carbon black (nCB), and single-layer graphene oxide (GO) on settlement of Amphibalanus amphitrite (Cirripedia, Crustacea) cypris larvae (cyprids) were assessed after 24, 48, and 72 h of exposure. Additionally, the effects of these nanomaterials on the mortality and swimming behaviour of the nauplius larvae (nauplii) of the same organism were determined after 24 and 48 h. The data indicate that nCB is more effective as a potential antisettlement agent than single-layer GO; moreover, nCB did not show any adverse effects on the larvae. The swimming behaviour of II stage nauplii of A. amphitrite exposed to a suspension of nCB was inhibited only at very high nCB concentrations (≥0.5 mg/mL). Single-layer GO, on the contrary, showed lower antisettlement effects and was more active in altering the survival and inhibiting the swimming behaviour of the nauplii. An indication of the toxic or non-toxic mechanisms of the antisettlement properties of both of these nanomaterials is provided by the reversibility of the antisettlement activity. In conclusion, we propose nCB as an innovative antifouling nanomaterial that shows low toxicity towards the model organism (crustaceans) used in this study.     

The influence of bacterial quorum-sensing inhibitors against the formation of the diatom-biofilm

Quorum-sensing inhibitor (QSI) is one of the most promising and environmentally friendly agents for marine antifouling. In this study, the activities of three kinds of QSIs 3,4-dibromo-2(5)H-furanone, 4-nitropyridine-N-oxide and indole were evaluated on the growth of two marine diatoms Cylinthrotheca sp. and Nitzschia closterium. At the same time, the effects of QSIs on the formation of the diatom-biofilm were also discussed. All the results showed that QSIs significantly inhibited the growth, and the effects depended on the dose and diatom species. The extracellular polymeric substance contents in the diatom-biofilm were significantly reduced by QSIs. However, the contents of polysaccharide in culture mediums were increased, which might result in the destruction of diatom cells. Combined with the results of crystal violet staining-biofilm and images of scanning electron microscopy, it was further demonstrated that QSIs inhibited the biofilm formation of Cylindrotheca sp., and the inhibitory effect of 4-nitropyridine-N-oxide was superior to that of 3,4-dibromo-2(5)H-furanone and indole.     

Decrease of net carbon flux in two species of mussels caused by extracts of crude oil

Effects of crude oil and salinity stress on the metabolism of two common filter-feeding animals (Mytilus edulis and Modiolus demissus) have been investigated. Carbon budgets have been calculated for each species under a variety of combinations of oil content and salinity. Both reduced salinity and the presence of crude oil tend to decrease the net carbon flux for each species; stresses from each source interacted in their effects on experimental animals. Although similar responses to oil were shown by each species, Mytilus edulis appeared to be slightly more resistant to oil than Modiolus demissus.     

Effects of organic additions to sediment,and maternal age and size,on patterns of offspring investment and performance in two opportunistic deposit-feeding polychaetes

The investment made by a reproducing organism in its offspring can be affected by both extrinsic (environmental) and intrinsic (female condition) factors. The purpose of this study, conducted between December 1990 and June 1992, was to examine the influence of organic additions to sediment, and female age and size, on patterns of offspring investment and performance in the opportunistic polychaete Capitella sp. I (cf. Capitella capitata Farbricius). Sediment treatments were composed of marsh mud enriched with sewage or algae, or spiked with #2 fuel oil. All three of these additions are associated with opportunistic responses in Capitella sp. I populations in the laboratory and field. As a means of comparison,effects of sewage additions were also examined on the spionid Streblospio benedicti Webster. Organic additions to mud had no detectable effect on the C or N content of embryos in either species. However, sewage had a positive effect on juvenile performance in Capitella sp. I. Juveniles produced by adult Capitella sp. I in the sewage treatment grew 2 times larger (in 2 wk) than juveniles produced by adults in the mud treatment when these juveniles were raised in the same sediment type (mud). Such effects may facilitate opportunistic responses in Capitella sp. I offspring by reducing time to first reproduction. In addition to treatment sediment effects, maternal age and size had a strong influence on patterns of offspring investment in Capitella sp. I. The material investiment measured in collected embryos declined with maternal age in the oil and sewage treatments. The C:N ratio of embryos increased with maternal age for Capitella sp. I except in the algae treatment, which showed no effect of maternal age. Juvenile growth decreased with maternal age in the sewage treatment for Capitella sp. I. Maternal size appeared to have a positive effect on embryo investment in the mud and oil treatments, where reproducing worms were smallest. Previous studies of Capitella sp. I have documented a broad range of environmentally induced responses in growth and fecundity.The results of this study emphasize the importance of maternal environment, age, and size effects on the relationship between offspring investment and performance.Such effects may have profound consequences for the dynamics of populations and the fitness of individuals. Present address: WES-ES-F, USAE Waterways Experiment Station, 3909 Halls Ferry Rd., Vicksburg, Mississippi 39180-6199, USA