Embryo aberrations in the amphipod Monoporeia affinis as indicators of toxic pollutants in sediments: A field evaluation

Reproduction in the amphipod Monoporeia affinis is sensitive to contaminant exposure, and embryo aberrations are used in the Swedish National Monitoring Program to indicate biological effects of contaminants. However, empirical support for a causal relationship between contaminant load and frequency of aberrations is largely based on experimental studies. A field evaluation of aberration frequency in relation to contaminant exposure is required, if we are to use reproductive disorders as indicators of contaminant effects in risk assessment. In this study, we evaluated occurrence of females carrying malformed embryos and frequency of specific embryo aberrations in broods of M. affinis in relation to concentrations of various pollutants in sediments. Gravid M. affinis and surface sediment were sampled at ten sites in the Gulf of Bothnia located at varying proximity to historical and current polluting point sources. At each site, concentrations of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and trace and heavy metals in surface sediments were determined. Amphipod fecundity and embryo development status were assayed with regard to morphologically recognizable aberrations (malformed embryos, membrane-damaged embryos, embryos with arrested development and females with dead brood). Our key finding is that different types of embryo aberrations were significantly associated with specific contaminant groups in the sediment. In particular, occurrence of females with embryo malformations was strongly related to elevated concentrations of Cd and PCBs, while females with membrane-damaged embryos occurred at high PAH concentrations. Also, frequency of embryos with arrested development was linked to elevated concentrations of PAHs and metals. Finally, frequency of females carrying dead broods, which was earlier considered to be a hypoxia-induced pathology, was significantly positively related to PAH concentrations. Thus, these reproductive aberrations in M. affinis can serve as contaminant-specific indicators of PCB, PAH and heavy metal exposure in biological effect monitoring.     

Seasonal variations of a battery of biomarkers and physiological indices for the mussel Mytilus galloprovincialis transplanted into the northwest Mediterranean Sea

Seasonal variations of six mussel (Mytilus galloprovincialis) biomarkers at two sites in the Mediterranean Sea were compared with physiological indices (condition, growth and gonad maturation), environmental parameters (temperature, salinity and turbidity), and chemical contamination levels. The basal levels of acetylcholinesterase (AChE), DNA adducts, benzo[a]pyrene hydroxylase (BPH), heat-shock proteins (HSP70), metallothioneins (MT) and P-glycoprotein (P-gp)-mediated multixenobiotic resistance (MXR) were estimated as early warning signals in caged mussels sampled at Carteau (native site) and La Fourcade (transplantation site) over a 2-year period. The Carteau and La Fourcade mussels have specific chemical contamination profiles but a similar range of values. For example, both are highly contaminated by heavy metals (201 and 258.4 mg kg(-1) dw, respectively) and considered as moderately impacted for polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs). However, contamination levels at Carteau are twice as high for PAHs (101.5 mg g(-1) dw) and PCBs (90.2 mg g(-1) dw) than La Fourcade. The seasonal contamination trend at Carteau showed six-fold higher levels of pyrolytic pollutants in winter. Although few tissue lesions were detected in individuals studied at either site, greater parasitic infestation was observed at Carteau. The results of findings from the two Mediterranean pilot studies support the adaptability of transplanted mussels to be used as biomarkers and to establish physiological endpoints for chemical contaminant exposure.     

Embryolethality and induction of 7-ethoxyresorufin O-deethylase in chick embryos by polychlorinated biphenyls and polycyclic aromatic hydrocarbons having Ah receptor affinity.

The lethality and 7-ethoxyresorufin O-deethylase (EROD)-inducing potency of some individual polycyclic aromatic hydrocarbons (PAHs) and coplanar polychlorinated biphenyls (PCBs) in chick embryos were measured in order to compare the mechanisms of action of these compounds. In previous studies it was found that coplanar PCBs and certain PAHs have a high embryolethality in the chicken and that they induce embryonic EROD activity. Although the most potent PAHs were almost as embryolethal as the PCBs when injected into hens' eggs 72 h prior to measurement, they were considerably less potent EROD inducers. In the present study, three coplanar PCBs (3,3',4,4'-tetrachlorobiphenyl (TCB), 3,3',4,4',5-pentachlorobiphenyl (PeCB) and 3,3',4,4',5,5'-hexachlorobiphenyl (HCB)) and four of the most toxic PAHs (benzo[a]anthracene (BaA), benzo[k]fluoranthene (BkF), indeno[1,2,3-cd]pyrene (IP) and dibenzo[a, h]-anthracene (DBahA] were administered to chick embryos in different ways, including co-administration. Additive embryolethality was found when BkF and PeCB were co-administered as well as when BaA and DBahA were given simultaneously. The PAHs were more effective as EROD inducers when injected on day 9 (24 h prior to measurement) than when injected on day 7 (72 h prior to measurement). The opposite was found for PeCB and HCB, whereas no difference in potency was noted when comparing TCB injected 24 and 72 h before EROD determination. These substance-related differences were probably due, at least partly, to differences in biotransformation rates. EROD activities found after treatment with high doses of BkF, IP, or DBahA on day 9 were similar to those measured after treatment with PeCB in doses high enough to give maximal induction. Co-administration of high doses of BkF and PeCB did not further increase the activity, indicating that the PAHs and coplanar PCBs induce EROD to a common maximal value. To decrease the influence of metabolization of the PAHs on their EROD-inducing potency, EROD was determined early in development (day 8) and soon after treatment (24 h) in one experiment. In that experiment, the PAHs proved to be only a few times less potent EROD inducers in relation to their embryolethalities compared with the PCBs. The results of the present study, a previously observed similarity in pathology between chick embryos treated with PAHs and embryos treated with coplanar PCBs, and the fact that the most toxic PAHs also are the most avid Ah receptor binders suggest that the coplanar PCBs and the PAHs largely exert their toxicity in chick embryos via an Ah receptor-mediated mechanism. The differences between the compounds in their EROD-inducing potency/embryolethality ratios could probably be explained by their different rates of biotransformation.     

Female fecundity and early offspring growth in the guppy, <Emphasis Type="Italic">Poecilia reticulata</Emphasis>

We examined the relationships between family (female parentage), body size of females, brood retention time between mating and parturition, female fecundity, and early growth of offspring in the guppy Poecilia reticulata. Mature, virgin females from a single brood were mated with a single male. Results of generalized linear models indicate that the effect of the family on female fecundity and offspring growth was significant, which suggested that these traits are genetically determined to a certain extent. Larger females at the time of mating produced larger broods, although female body size at the time of parturition did not affect brood size, in contrast to the results of some previous studies in guppies. Brood size was negatively associated with the body size of neonates. Results highlighted significant associations between brood retention time and female fecundity as well as offspring growth. In addition, the interaction between the family and brood retention time was significantly associated with female fecundity and offspring growth. Females of some families had longer retention times of larger broods, whereas those of other families had shorter retention times of smaller broods. On the other hand, females with longer brood retention times produced smaller neonates with slower growth. Since the family also affected the brood retention time, selection may work against the duration of brood retention of females via the size, growth and number of offspring, depending on environmental factors such as the intensity of predation or competition in neonates.     

A review of the reproductive bionomics of aquatic gammaridean amphipods: variation of life history traits with latitude,depth, salinity and superfamily

Life history traits (mean and maximum body length of females, number of embryos per brood = brood size, embryo diameter, number of broods per female, lifespan of females) for 302 populations of aquatic gammaridean amphipods, representing 214 species in 16 superfamilies, were reviewed. The variation of these traits, of lifetime potential fecundity (i.e. the number of embryos produced per female lifespan) and of reproductive potential (i.e. the number of embryos produced per female per year), with temperature (latitude), depth, salinity and superfamily, was investigated by various univariate and multivariate methods. Gammaridean amphipods comprise semelparous and iteroparous populations and species, with semiannual, annual, biannual or perennial life cycles. However, most gammarideans studied so far are iteroparous annuals. Body length explains most of the variation in brood size and embryo diameter. The reproductive potential may be increased by increasing body size for a constant breeding frequency, by increasing brood size at the expense of smaller embryos, by increasing breeding frequency for a constant lifespan at the expense of smaller individual broods and/or embryos, and by increasing longevity for a constant breeding frequency and brood size. Combinations of these different options constitute the life history patterns of gammarideans, which vary across superfamilies, latitude and depth, and cannot simply be explained by variations in body length. High latitude species were generally characterized by biannual or perennial life histories, large body size, delayed maturity, and single or few broods with many, relatively large embryos; converse sets of traits characterized low latitude species. Deep-living species had relatively smaller broods and embryos than their shallow-living relatives, yet did not produce more broods. However, different superfamilies dominated in different habitats. The importance of natural selection relative to phylogenetic (historical) and physiological constraints in the forging of these patterns is discussed.     

An Integrated Case Study for Evaluating the Impacts of an Oil Refinery Effluent on Aquatic Biota in the Delaware River: Introduction,Study Approach,and Objectives

This is the first in a series of article presenting results from a case study designed to assess the impacts of an oil refinery effluent [primarily polynuclear aromatic hydrocarbons (PAHs)] on aquatic biota in the Delaware River. During the course of the study, the oil refinery was owned by Motiva Enterprises LLC. This article provides background information on the study area, the study approach and objectives. The specific objectives of this multiyear study were to: (1) measure water column concentrations of PAHs and other contaminants (i.e., metals) in Motiva's effluent and intake canal and selected Delaware River sites; (2) assess fate and transport issues associated with the Refinery effluent; (3) characterize sediment PAHs, total organic carbon (TOC), and grain size distributions in the discharge canal, near-field, mid-field and far-field areas of the Refinery to aid in the selection of Triad sample sites (including reference areas); (4) conduct Triad studies (chemical characterizations, sediment toxicity assessments, and benthic community characterizations) at selected study sites during the spring and summer of 2001 and 2002; (5) perform fingerprinting of PAHs in Motiva's effluent to differentiate Motiva-related PAHs in sediment and biota from other sources; (6) assess bioavailability of PAHs, PCBs, and metals by using resident bivalve studies; (7) conduct long-term coring to determine potential impact of past non-complying discharges; and (8) integrate and analyze all study components to address the research goals. The results from objectives 1, 2, and 3 are briefly summarized in this series of articles whereas the other five objectives are the subject of the various papers presented in this volume.     

Plant terpenes and lignin as natural cosubstrates in biodegradation of polyclorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs)

The objective of this minireview is to examine how cometabolic biodegradation of polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) might be affected by plant terpenes and lignins as natural substrates abundant in nature. The topics covered, hence, are environmental significance of PCBs and PAHs, nature and distribution of plant terpenes and lignin, structural and metaoblic similarities of the natural compounds to PCBs and PAHs, and possible roles of the natural substrates in inducing the biodegradative pathways of PCBs and PAHs.     

Sucralose Induces Biochemical Responses in Daphnia magna

The intense artificial sweetener sucralose has no bioconcentration properties, and no adverse acute toxic effects have been observed in standard ecotoxicity tests, suggesting negligible environmental risk. However, significant feeding and behavioural alterations have been reported in non-standard tests using aquatic crustaceans, indicating possible sublethal effects. We hypothesized that these effects are related to alterations in acetylcholinesterase (AChE) and oxidative status in the exposed animals and investigated changes in AChE and oxidative biomarkers (oxygen radical absorbing capacity, ORAC, and lipid peroxidation, TBARS) in the crustacean Daphnia magna exposed to sucralose (0.0001–5 mg L⁻¹). The sucralose concentration was a significant positive predictor for ORAC, TBARS and AChE in the daphnids. Moreover, the AChE response was linked to both oxidative biomarkers, with positive and negative relationships for TBARS and ORAC, respectively. These joint responses support our hypothesis and suggest that exposure to sucralose may induce neurological and oxidative mechanisms with potentially important consequences for animal behaviour and physiology.     

Poecilogony in the caenogastropod Calyptraea lichen (Mollusca: Gastropoda)

In marine invertebrates, polymorphism and polyphenism in mode of development are known as “poecilogony.” Understanding the environmental correlates of poecilogony and the developmental mechanisms that produce it could contribute to a better understanding of evolutionary transitions in mode of development. However, poecilogony is rare in marine invertebrates, with only ten recognized, well‐documented cases. Five examples occur in sacoglossan gastropods, and five occur in spionid polychaetes. Here, we document the eleventh case, and the first in a caenogastropod mollusc. Females of Calyptraea lichen collected in the field or reared in the laboratory often produce broods of planktotrophic larvae. They can also be collected with mixed broods, in which each capsule contains planktotrophic larvae, nurse embryos, and adelphophagic embryos. Adelphophages eat the nurse embryos and hatch as short‐lived lecithotrophic larvae, or even as juveniles. Mitochondrial COI and 16S DNA sequences for females with different types of broods differ by less than 0.5%, supporting conspecific status. Some females collected in the field with mixed broods subsequently produced planktotrophic broods, demonstrating that females can produce two different kinds of broods. Calyptraea lichen is therefore polyphenic in two ways: mode of development can vary among embryos within a capsule, and females can change the types of broods they produce.     

Scale of classification based on biochemical markers in mussels: application to pollution monitoring in European coasts

A battery of biochemical parameters is used to evaluate the response of mussels to a contaminated coastal environment. In the European BIOMAR programme, a multimarker approach was developed, establishing a scale for the classification of the water quality in European coastal sites. AChE activity is highly sensitive to organophosphorus and carbamate insecticides but also to heavy metals. Catalase activity and lipid oxidation (evaluated as MDA) are markers of oxidative stress, GST activity is related to the conjugation of organic compounds and BPH activity is a marker of planar compounds (e.g. PAHs). These parameters were measured either in gills (AChE, GST) or digestive gland (BPH, GST, CAT, MDA). Contamination levels were estimated by measurement of PAHs and heavy metals in animals. For each biomarker, a discriminatory factor was calculated (maximum variation range/confidence interval) and a response index was allocated. For each site, a global response index was calculated as the sum of the response index of each of the five biomarkers. As a result of our calculation method, the quality of the coastal environment at each site can be classified with a five level scale. Mussels were collected during five cruises in 1995-1996 on the Baltic and Mediterranean coasts. The results show that water quality ranged from class 1 (clean areas in some sites of the French Riviera, Spanish Costa Brava and the Baltic coast) to class 5 (high pollution in main harbours, e.g. Kiel and Toulon). Some areas fall into class 4, e.g. Carteau, Cortiou, Barcelona, Warnemunde, Swinemunde, Ebro delta. The global Biomarker Index was positively correlated with the level of PAHs in mussels in Baltic transects. A number of other contaminants or stressors may be present in the marine environment and the Biomarker Index appeared to be relevant to classify coastal environmental pollution.     

Scale of classification based on biochemical markers in mussels: application to pollution monitoring in European coasts

A battery of biochemical parameters is used to evaluate the response of mussels to a contaminated coastal environment. In the European BIOMAR programme, a multimarker approach was developed, establishing a scale for the classification of the water quality in European coastal sites. AChE activity is highly sensitive to organophosphorus and carbamate insecticides but also to heavy metals. Catalase activity and lipid oxidation (evaluated as MDA) are markers of oxidative stress, GST activity is related to the conjugation of organic compounds and BPH activity is a marker of planar compounds (e.g. PAHs). These parameters were measured either in gills (AChE, GST) or digestive gland (BPH, GST, CAT, MDA). Contamination levels were estimated by measurement of PAHs and heavy metals in animals. For each biomarker, a discriminatory factor was calculated (maximum variation range/confidence interval) and a response index was allocated. For each site, a global response index was calculated as the sum of the response index of each of the five biomarkers. As a result of our calculation method, the quality of the coastal environment at each site can be classified with a five level scale. Mussels were collected during five cruises in 1995-1996 on the Baltic and Mediterranean coasts. The results show that water quality ranged from class 1 (clean areas in some sites of the French Riviera, Spanish Costa Brava and the Baltic coast) to class 5 (high pollution in main harbours, e.g. Kiel and Toulon). Some areas fall into class 4, e.g. Carteau, Cortiou, Barcelona, Warnemunde, Swinemunde, Ebro delta. The global Biomarker Index was positively correlated with the level of PAHs in mussels in Baltic transects. A number of other contaminants or stressors may be present in the marine environment and the Biomarker Index appeared to be relevant to classify coastal environmental pollution.     

Superfetation increases total fecundity in a viviparous fish regardless of the ecological context

Superfetation is the ability of females to simultaneously carry multiple broods of embryos at different developmental stages. This is an uncommon reproductive strategy that has evolved independently several times in viviparous fishes. The ecological conditions that favor higher degrees of superfetation (the presence of more simultaneous broods) still remain unclear. In this study we tested hypotheses about the potential effects of three particular ecological factors (water flow velocity, population density, and adult mortality) on superfetation. We used data on six populations of one fish species from the family Poeciliidae (Poeciliopsis baenschi) and a multimodel inference framework to test these hypotheses. We found no clear associations between the degree of superfetation and these ecological factors. Instead, we found a positive relationship between the total number of embryos carried by females and superfetation. Females increased their total fecundity as they overlapped more broods and this pattern was independent of the particular ecological conditions. Thus, in P. baenschi superfetation may facilitate a greater reproductive output. In addition, this positive relationship between total number of embryos and superfetation was stronger in small- and medium-sized females, whereas large females produced few or no simultaneous broods regardless of their total fecundity. The observed lack of association between superfetation and ecological variables is noteworthy because previous studies on other congeneric species have found that superfetation may vary as a function of water flow velocity or food availability. Our results indicate that the effect of particular selective factors on the degree of superfetation may differ among closely related species.     

Transfer of microcontaminants from sediment to chironomids, and the risk for the Pond bat Myotis dasycneme (Chiroptera) preying on them

Transfer ratios of metals, PCBs, pesticides and PAHs from the sediment to chironomid larvae and adults collected in a highly contaminated area, the Biesbosch, were studied. Metal concentrations in larvae were 0.28 (Cd), 0.02 (Cr), 0.52 (Cu), 0.06 (Hg), 0.03 (Pb), 0.32 (Zn) times those found in standard sediment, on a dry weight basis. Hg and Zn were well transported to the adult stage. Dry weight ratios of contaminant residues in adults and in larvae were 0.38 (Cd), 0.23 (Cr), 0.62 (Cu), 1.03 (Hg), 0.08 (Pb), 0.94 (Zn). For PCBs and pesticides, the concentration ratios of chironomid larvae fat to sediment (dry organic matter) varied around 3.3, which is consistent with laboratory studies. Organochlorine residues in adult fat were comparable to those in larvae lipids. However, the concentrations of non-ortho PCBs were 1.7 times higher in adults. For polycyclic aromatic hydrocarbons (PAH), larval fat:sediment (organic matter) concentration ratios ranged from 0.004 to 0.1. Adult: larvae ratios for PAHs varied between 0.2 and 0.6. For naphthalene a much higher transport ratio of 2 was found. Chironomid adults are the most important potential food source of the Pond bat, which lives in low densities in the Biesbosch. The contaminant concentrations measured in the chironomids do not exceed diet levels that are thought to be safe for mammals. However, Pond bats collected in less contaminated areas contained PCB-concentrations of 9, 33 and 76 mg kg^–1 lipid weight, which are above concentrations that cause reproduction effects on Mink.     

“Survival in air” of the blue mussel Mytilus edulis L. as a sensitive response to pollution-induced environmental stress

Mussels, Mytilus edulis, were exposed for periods of 6 weeks at various locations in Dutch coastal waters during 1989 and 1990. “Survival in air” showed to be a sensitive response parameter for indicating pollution induced environmental stress in transplanted mussels sampled from eight field sites. Increased tissue contaminant levels, especially PCBs and PAHs, correlated with a reduced survival time during aerial exposure. Three weeks exposure of mussels in the laboratory to 1 μg · 1⁻¹ PCBs affected the aerial survival time negatively. Laboratory experiments did not indicate that lowered salinity influences the “Survival in air” response after sufficient acclimation (15 days), facilitating the use of this response parameter in both marine and estuarine waters.     

In situ phytoremediation of PAH- and PCB-contaminated marine sediments with eelgrass (Zostera marina)

In view of the fact that there are presently no cost-effective in situ treatment technologies for contaminated sediments, a 60-week-long phytoremediation feasibility study was conducted in seawater-supplied outdoor ponds to determine whether eelgrass (Zostera marina) is capable of removing polynuclear aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) from submerged marine sediments. It was determined that all PAHs and PCBs, independent of the number of aromatic rings and degree of chlorination, respectively, were removed to a much larger extent in planted sediments compared to unplanted controls. After 60 weeks of treatment, the concentration of total PAHs decreased by 73% in planted sediments but only 25% in unplanted controls. Similarly, total PCBs declined by 60% in the presence of plants while none were removed in the unplanted sediment. Overall, the apparent PAH and PCB biodegradation was greatest in the sediment layer that contained most of the eelgrass roots. Abiotic desorption tests conducted at week 32 confirmed that the phytoremediation process was not controlled by mass-transfer or bioavailability limitations since all PAHs and PCBs desorbed rapidly and to a large extent from the sediment. PAHs were detected in both roots and shoots, with root and shoot bioaccumulation factors for total PAHs amounting to approximately 3 and 1, respectively, after 60 weeks of phytoremediation treatment. Similarly, the root bioaccumulation factor for total PCBs was around 4, while no PCBs were detected in the eelgrass leaves at the end of the experiment. The total mass fraction of PAHs and PCBs absorbed and translocated by plant biomass during the 60-week period was insignificant, amounting to less than 0.5% of the total mass of PAHs and PCBs which was initially present in the sediment. Finally, the number of total heterotrophic bacteria and hydrocarbon degraders was slightly but not statistically significantly greater in planted sediments than in unplanted controls. After ruling out contaminant loss to the water column or absorption and transformation within plant cells, it is most likely that the presence of eelgrass stimulated the microbial biodegradation of PAHs and PCBs in the rhizosphere by releasing root exudates, plant enzymes, or even oxygen. Additional research is needed to further elucidate these potential phytoremediation mechanisms.     

Developmental inhibition and reproductive potential impairment in Musca domestica L. by heavy metals

Post embryonic development and adult emergence of housefly, M. domestica L. was adversely affected in varying degrees on exposure of larvae to 6 different heavy metals. Of these, salts of mercury, aluminium and cadmium exhibited significant reduction in normal adult emergence. High decline in reproductive potential in terms of female fecundity and egg hatchability was also observed in the F1 progenies of treated larvae. The results are discussed in light of heavy pollution of environment by such contaminants.     

Persistent organic pollutants in the blood of free-ranging sea otters (Enhydra lutris ssp.) in Alaska and California

As part of tagging and ecologic research efforts in 1997 and 1998, apparently healthy sea otters of four age-sex classes in six locations in Alaska and three in California were sampled for persistent organic pollutants (POPs) and other chemicals of ecologic or environmental concern (COECs). Published techniques for the detection of POPs (specifically ∑polychlorinated biphenyls [PCBs], ∑DDTs, ∑hexachlorocyclohexanes [HCHs], ∑polycyclic aromatic hydrocarbons [PAHs], ∑chlordanes [CHLs], hexachlorobenzene [HCB], dieldrin, and mirex) in the tissue of dead otters were modified for use with serum from live sea otters. Toxic equivalencies (TEQs) were calculated for POPs with proven bioactivity. Strong location effects were seen for most POPs and COECs; sea otters in California generally showed higher mean concentrations than those in Alaska. Differences in contaminant concentrations were detected among age and sex classes, with high levels frequently observed in subadults. Very high levels of ∑DDT were detected in male sea otters in Elkhorn Slough, California, where strong freshwater outflow from agricultural areas occurs seasonally. All contaminants except mirex differed among Alaskan locations; only ∑DDT, HCB, and chlorpyrifos differed within California. High levels of ∑PCB (particularly larger, more persistent congeners) were detected at two locations in Alaska where associations between elevated PCBs and military activity have been established, while higher PCB levels were found at all three locations in California where no point source of PCBs has been identified. Although POP and COEC concentrations in blood may be less likely to reflect total body burden, concentrations in blood of healthy animals may be more biologically relevant and less influenced by state of nutrition or perimortem factors than other tissues routinely sampled.     

Experimental dissociation of individual quality, food and timing of breeding effects on double-brooding in a migratory songbird

Annual reproductive success in many species is influenced by the number of breeding attempts within a season. Although previous studies have shown isolated effects of female quality, food, and timing of breeding on the probability of female birds producing second broods, to our knowledge, none have tested the relative importance of multiple factors and their interactions using simultaneous manipulations within populations of free-living birds. In this study, we show that individual quality and timing of breeding interact to affect the probability of double-brooding in female mountain bluebirds (Sialia currucoides). High-quality females (those that naturally initiated clutches early in the season) were more likely to double-brood, regardless of whether their hatching date was advanced or delayed, whereas later breeding, lower quality females were much less likely to double-brood when their first attempt was delayed. This indicates that annual fecundity of poorer quality (or younger) female bluebirds may be more sensitive to seasonal variation in environmental conditions. In addition, birds that were provided with supplemental food throughout first breeding attempts were more likely to double-brood in one of the study years, suggesting that female bluebirds may be energetically limited in their capacity to initiate a second brood. Females that had their first brood delayed also had a shorter inter-brood interval and were moulting fewer feathers during second broods compared to controls, while females in better condition showed more advanced moult in second breeding attempts. Taken together, our results demonstrate the combined effects of age- or individual quality-mediated energetic trade-offs between current and future reproduction, and between investments in offspring and self-maintenance, on annual fecundity of female birds.     

Reproductive behavior of intersexes of an intertidal amphipod Corophium volutator

Abstract. Intersexes are common in crustaceans. Typically, these intersexes are sterile or function as females, but prior evidence from laboratory experiments suggests that intersexes of a key species of gammaridean amphipod, Corophium volutator , might function as males. We observed that intersexes of C. volutator behaved as males by crawling (mate-searching) on a mudflat during ebb tides and pairing in burrows with female amphipods. In the laboratory, intersexes and males did not differ in aspects of crawling such as movement rate and measures of burrow investigation. I`ntersexuality was costly in that intersexes crawled less often than males on a mudflat, formed fewer pairs with females than males, and remained in tandem less often with receptive females than males. The use of PCR-based identification methods failed to identify the presence of transovarial, feminizing, microsporidian parasites as a major cause of intersexuality in this species in that infected females did not produce broods that contained more intersexes than broods produced by uninfected females. Because intersexes may be mistaken as females, the percentage of functional males in amphipod populations may be underestimated: an important consideration given male limitation in populations of C. volutator. The occurrence of intersexes has significant implications for studies on the evolution and ecology of sex ratios, and the use of crustaceans as indicators of environmental quality.     

Biomarkers in Zebra mussel for monitoring and quality assessment of Lake Maggiore (Italy)

Three different biomarkers (acetylcholinesterase (AChE), ethoxy resorufin-O-deethylase (EROD) and DNA strand breaks) were measured in Zebra mussel (Dreissena polymorpha) specimens collected in April 2005 at six different sampling sites on Lake Maggiore, the second largest Italian lake in terms of depth and volume, in order to assess the spatial variation of exposure to man-made contaminants. Mussels maintained at fixed laboratory conditions were used as controls to eliminate potential interference due to environmental factors. Biomarker data were also supported by the analysis of several chemicals (six dichlorodiphenyltrichloroethane (DDTs), 23 polychlorinated biphenyls (PCBs), 14 polybrominated diphenyl ethers (PBDEs), 11 polycyclic aromatic hydrocarbons (PAHs) and hexachlorobenzene (HCB)) measured in the mussel soft tissues by gas chromatographic analyses. We found a negative correlation between temperature and AChE activity, while any measured environmental or physiological factor seemed to influence EROD activity and DNA strand breaks. A positive relationship was found between EROD activity and all of the measured chemicals, except for PAHs, which correlated with the amount of DNA strand breaks. Significant differences were noted for all biomarkers, both among sampling stations and between control and experimental data, even if the general level of variability was low. The biomarkers showed a distinct pattern of spatial variation, but the evaluation of DNA strand breaks was the strongest discriminating power between sites. In addition, the comparison between AChE and EROD activity measured in 2005 was compared with results obtained in a previous study carried out over the same sampling period in 2003. Results indicated a strong influence of temperature on AChE activity and probable interference of substrate inhibition of EROD activity, pointing out the need to take care in the interpretation of data comparisons. The results obtained with two different metrics used for the measure of DNA strand breaks is also discussed, as well as the relationship between EROD activity data and potential genotoxicity.