Comparison of supercritical fluid extraction and Soxhlet extraction for the determination of PCBs in seaweed samples

The efficiency of supercritical fluid extraction for the determination of 12 polychlorinated biphenyls from algae samples is compared to Soxhlet extraction. Analytical detection limits for the individual congeners ranged from 0.62 microgl(-1) to 19 microgl(-1). Recovery was tested for both methods using standard addition procedure. At maximum spike level of concentration, the mean recoveries were not significantly different (P>0.05) of all PCBs studied, with the exception of PCBs 28, 52, 77 and 169. Method precision for Soxhlet extraction (< or =3.9%) was slightly better than for SFE (< or =9.2%). Although both methods yield comparable results, SFE offers the advantage of detecting all PCBs studied at lower concentrations, reducing extraction time, and reducing the amount of solvents needed. The optimized methods were applied to the analysis of three real seaweed samples, except for PCB101 the concentrations of all PCBs were low or below the detection limits. The levels of PCB101 found in sample 1 were 6.6+/-0.54 ng g(-1) d.w., in sample 2 the levels were 8.2+/-0.86 ng g(-1) d.w. and in sample 3 they were 7.7+/-0.08 ng g(-1) d.w.     

Health-dependent vulnerability to predation affects escape responses of unguarded chinstrap penguin chicks

Predators may select more often to attack the more vulnerable prey or those with an inferior health status. Thus, prey should be able to assess their own vulnerability to predation and modify their antipredatory behavior accordingly. When approached by predator skuas, unguarded penguin chicks flee short distances, and usually aggregate in dense packs, but there is a clear interindividual variability in their responses under similar conditions. We hypothesized that this variability in escape responses might be related to the perceived vulnerability to predation of each individual chick. We simulated predator attacks to chinstrap penguin chicks and analyzed the sources of variation in their escape response, such as the presence of adults or the density of other chicks, and the sex, age, body condition, and health status of responding chicks. Chicks allowed shorter approach distances when they had a better health condition (i.e., a greater T-cell-mediated immunity, CMI), when they were younger, and when the density of adults around was higher. Sex and density of other chicks were not important. Similarly, chicks fled from the experimenter to longer distances when they had a lower CMI and when the density of adults was lower. Therefore, escape characteristics of chicks depended on the presence of adults that can deter predators and on the health-dependent vulnerability of chicks.     

Yolk androgen deposition as a compensatory strategy

The deposition of androgens into the yolks of eggs can have long-lasting effects on the growth and development of young birds. It has been proposed that female birds and reptiles deposit yolk androgens according to the differential allocation hypothesis (DAH), which posits the allocation of more resources to offspring sired by more attractive, higher-quality males. We examined deposition patterns of yolk androgens in relation to mate attractiveness in the house finch Carpodacus mexicanus. Contrary to the predictions of the DAH, female house finches deposited significantly more androgens into eggs sired by less attractive males. We propose that, rather than serving as resources, androgens are used as mediators in a compensatory distribution strategy, enabling females to improve the quality of young produced with less attractive males.     

Genome-size variation in bivalve molluscs determined by flow cytometry

Six of the nine described species of the mole crab genus Emerita are distributed in the Americas, two [E. analoga (Stimpson, 1857) and E. rathbunae Schmitt, 1935] on the west coast, and four [E. benedicti Schmitt, 1935, E. brasiliensis Schmitt, 1935, E. portoricensis Schmitt, 1935 and E. talpoida (Say, 1817)] on the east. The presence of an extended planktonic larval stage in all Emerita species suggests high dispersal potential and the possibility of extensive gene flow among conspecific populations. Two taxa were sampled to study the extent of gene flow between widely separated conspecific populations: E. analoga (California and Chile) and E. talpoida (Massachusetts, South Carolina, and the west coast of Florida), while all other taxa were characterized from a single location. Portions of two mitochondrial genes, cytochrome oxidase I (COI) and 16S ribosomal RNA (16S rRNA) were sequenced. For data analysis, approximately 500 bp (COI) and 400 bp (16S rRNA) were examined. Estimated genetic divergence of 5.41% in COI between E. talpoida populations sampled from the Gulf of Mexico and the Atlantic coast, and 3.47% between E. analoga sampled in Chile and California, indicates that in both cases there has been no recent gene flow between disjunct populations. Additional molecular and morphological studies are necessary to decide whether disjunct populations should be accorded specific status. We predict that many marine invertebrates with antitropical distributions similar to E. analoga may consist of sibling species. In contrast to relationships inferred earlier from distribution patterns, parsimony analyses of both COI and 16S rRNA data yield similar phylogenetic trees in which E. analoga is separated from a clade composed of other species in the Americas; a bootstrap value (67%) in the COI inferred tree marginally supports the separation, but the same tree topology with a higher bootstrap value (84%) is obtained with 16S rRNA sequence data. Genetic divergence among the taxa indicates that the Emerita species constitute an old group and that distribution of species has been modified by past climatic and geological events.     

Abnormal gonadal morphology in intersex,<Emphasis Type="Italic"> Echinogammarus marinus</Emphasis> (Amphipoda): a possible cause of reduced fecundity?

Recent reports have demonstrated a cost associated with intersexuality in Amphipoda, including reduced fecundity and fertility. In this study, the gross morphology of the gonads in normal and intersex Echinogammarus marinus (Amphipoda) were compared to determine whether resource allocation to gonadal tissue accounted for this reduced fitness. Evidence for the presence of the male sex-determining hormone, androgenic gland hormone (AGH), was compared between sexual phenotypes using MALDI mass spectrometry. Two distinct intersex phenotypes (male intersex and female intersex) were found, with variation in gonadal structure corresponding with external phenotype. Examination of male intersexes revealed normal testicular development (testes, seminal vesicles and vas deferens), but also revealed the formation of an oviduct. Ovaries of intersex females showed normal ovarian development, but were reduced in length by approximately 20% due to the presence of vas deferens. The number of vas deferens in intersex females was equal to the number (one or two) of genital papillae. We hypothesise that the reduced ovarian length observed in intersex females is a likely cause of the reduced brood size previously reported in intersex females of this species. Variation in the sexual phenotype corresponded both to development of the androgenic gland and to expression of a peptide fragment corresponding to the A chain of androgenic gland hormone (AGH). Androgenic glands and a putative AGH peptide were present in males. However, in both normal and intersex females, the androgenic glands were only present in a rudiment form and the peptide was not detected. Intersex males were found to possess abnormal glands that appeared hypertrophied. However, AGH peptides were not detected, supporting the suggestion that the intersex phenotype is manifested via perturbations of AGH.An erratum to this article can be found at Communicated by J.P. Thorpe, Port Erin     

Removal of the herbicide amitrole from water by anodic oxidation and electro-Fenton

Here we demonstrate that an aqueous solution of the herbicide amitrole can be completely depolluted at pH 3.0 by anodic oxidation and electro-Fenton process. Anodic oxidation gives faster degradation with a boron-doped diamond anode than with a Pt anode. Electro-Fenton with a Pt anode and 1 mmol l ^–1 Fe²⁺ as catalyst yields the quickest depollution. Amitrole decay always follows a pseudo first-order reaction. NO₃^– and NH₄⁺ are accumulated in the medium during mineralization, although volatile N-products are also formed. These environmentally friendly electrochemical treatments could be applied to the remediation of wastewaters containing amitrole.     

Regulation of fructose-2,6-bisphosphate content in mantle tissue of the sea musselMytilus galloprovincialis

6-phosphofructo-2-kinase (PFK-2) from the mantle of the sea musselMytilus galloprovincialis Lmk, collected from the Ría de Arosa (NW Spain) in 1990, was purified 550-fold by extraction and sequential affinity chromatography on Affi-gel Blue and ATP-agarose columns. The enzyme was a dimer with a native molecular weight of 100 kilodaltons (KDa) and a subunitM _r of 53 KDa. PFK-2 activity is dependent on the presence of P_i. At physiological P_i concentrations, the enzyme exhibits hyperbolic kinetics with both ATP and Fru-6-P, withK _m values of 0.62 and 0.37 mM respectively. In vivo, PFK-2 activity is limited by the concentration of Fru-6-P which is low in comparison with theK _m for this substrate. Citrate and PEP inhibited PFK-2 activity.     

Chloroacetic acids in environmental processes

The fate of chloroacetic acids (CAA) in forest soils was studied using radio-indicator methods. We showed that chloroacetic acids are both microbially degraded and simultaneously formed by chloroperoxidase-mediated chlorination of acetic and humic acids. The degree of biodegradation of chloroacetic acids in soil depends on their concentration. Dichloroacetic acid (DCA) is degraded faster than trichloroacetic acid (TCA). Chlorination of acetic acid led to a fast formation of dichloroacetic acid, whereas chlorination of humic acids gave rise to trichloroacetic acid. Both processes lead to a steady state in soil, participate in the chlorine cycle and possibly also in decomposition of organic matter in forest ecosystems.     

Desperado siblings: uncontrollably aggressive junior chicks

According to the desperado sibling hypothesis, chicks of obligately siblicidal species kill their junior broodmates as early as possible because junior broodmates face dire ecological prospects and are consequently predisposed to aggressively overthrow elder broodmates by all-out aggression. The agonistic behavior of junior chicks is virtually unknown because they die so young. We investigated the agonistic behavior of hypothetical desperados by fostering junior brown boobies (Sula leucogaster, an obligately siblicidal species) into nests of the blue-footed booby (S. nebouxii, a facultatively siblicidal species) containing an older singleton blue-foot chick. Controls were junior blue-footed boobies fostered into the same situation. Junior brown boobies were 7 times more aggressive than controls and most of them dominated their elder and larger nestmates. Four of nine brown booby juniors showed relentless aggression, delivering up to 711 pecks, bites and pushes (including "expulsion pushes") per hour, thereby overwhelming nestmates 90% heavier and permanently expelling one of them from the nest. Similarly, in natural broods of two surviving brown booby chicks, the losing chick was 13 times as aggressive as blue-foot subordinates, up to at least age 7 weeks. Contrast of the two species of booby suggests the evolution of agonistic roles within broods may be partly driven by selection on potential victims to express a level and type of aggressiveness appropriate to their status-related ecological prospects.     

Copulatory plugs do not assure high first male fertilisation success: sperm displacement in a lizard

Sperm competition selects for opposing male defensive and offensive reproductive traits, and its outcome may be determined by the effectiveness to which one trait has evolved to out-compete the other. We tested the effectiveness of a first male plug physical interference with a second male insemination (defence) vs the effectiveness of plug and associated sperm displacement by a second male (offence) on the outcome of sperm competition in Iberian rock lizards. We conducted a double mating experiment where we compared the proportion of eggs per clutch fertilised by the same second males (against the same first males) when they copulated with females 30 min (first male plug adhered firmly inside the female cloaca) and 4 h (first male plug loosely adhered or shed from the female cloaca) after first males. We found that second males fertilised the majority of the eggs per clutch in the 30-min treatment, whereas fertilisations were equally shared between the two males in the 4-h treatment. These results show that plugs have little defensive effectiveness, and thus, do not assure high first male fertilisation success. Instead, sperm displacement appears to be associated with plug displacement. That is, because sperm embedded in first male plugs, and displaced from competition for fertilisations by second males, is expected to increase in number with decreasing time allowed for female sperm transport, second males thus enjoy higher fertilisation success. This study shows that offensive plug displacement out-competes plug defensive role in Iberian rock lizards. Moreover, it reveals sperm displacement as a novel sperm competition mechanism in reptiles.