Chemical defence in chewing and sucking insect herbivores: Plant-derived cardenolides in the monarch butterfly and oleander aphid

Summary Cardenolide sequestration by a hemimetabolous aphid and a holometabolous butterfly from the neotropical milkweed,Asclepias curassavica L., is compared. The oleander aphid,Aphis nerii B. de F., sequestered a similarly narrow range of cardenolide concentrations to the monarch butterfly,Danaus plexippus (L.), from the wide range of concentrations available in leaves of A.curassavica. However, A.nerii sequestered significantly less cardenolide (269 µg/0.1 g) thanD. plexippus (528 µg/0.1 g). The honeydew excreted by A.nerii was comprised of 46% cardenolide. The complete polarity range of 25 cardenolides detected by thin layer chromatography in A.curassavica was represented in the 17 whole aphid cardenolides and the 20 aphid honeydew cardenolides detected. D.plexippus sequestered a narrower polarity range of 11 cardenolides, having eliminated low polarity cardenolide genins and glycosides. It is suggested that these chemical differences may be related to interactions among the broad feeding tactics of sucking or chewing milkweed leaves, life history constraints of holometabolyversus hemimetaboly, the distribution of milkweed food resources in space and time, and the dynamics of natural enemies.     

Survival of first instar larvae ofDanaus plexippus (Lepidoptera: Danainae) in relation to cardiac glycoside and latex content ofAsclepias humistrata (Asclepiadaceae)

Summary Our paper addresses field survivorship of first instar monarch butterfly larvae (Danaus plexippus L., Lep.: Danainae) in relation to the dual cardenolide and latex chemical defenses of the sand hill milkweed plant,Asclepias humistrata (Asclepiadaceae) growing naturally in north central Florida. Survival of first instar larvae in the field was 11.5% in the first experiment (15–20 April 1990), and dropped to 3.4% in the second experiment (20–30 April). About 30% of the larvae were found glued to the leaf surface by the milkweed latex. Predator exclusion of non-flying inverte-brates by applying tanglefoot to the plant stems suggested that the balance of the mortality was due to volant inverte-brates, or to falling and/or moving off the plants. Regression analyses to isolate some of the other variables affecting survivorship indicated that first instar mortality was correlated with (1) increasing cardiac glycoside concentration of the leaves, (2) increasing age of the plants, and (3) the temporal increase in concentration of cardiac glycosides in the leaves. The study also provided confirmatory data of previous studies that wild monarch females tend to oviposit onA. humistrata plants containing intermediate concentrations of cardiac glycosides. Cardiac glycoside concentration in the leaves was not correlated with that in the latex. The concentration of cardenolide in the latex is extremely high, constituting an average of 1.2 and 9.5% of the mass of the wet and dry latex, respectively. The data suggest that an increase in water content of the latex is compensated for by an influx of cardenolide with the result that the cardenolide concentration remains constant in the latex systems of plants that are growing naturally. We also observed first instar larvae taking their first bite of milkweed leaves in the field. In addition to confirming other workers findings that monarch larvae possess elaborate sabotaging behaviour of the milkweed's latex system, we discovered that several larvae on their first bite involuntarily imbided a small globule of latex and instantly became cataleptic. This catalepsis, lasting up to 10 min, may have been in response to the high concentration of cardenolide present in the latex ofA. humistrata, more than 10 times that in the leaves. The results of the present study suggest that more attention should be directed to plant chemical defenses upon initial attack by first instar insect larvae, rather than attempting correlations of plant chemistry with older larvae that have already passed the early instar gauntlet. The first bite of neonate insects may be the most critical moment for coping with the chemical defenses of many plants and may play a much more important role in the evolution of insect herbivory than has previously been recognized.     

Increased nitrogen availability influences predator–prey interactions by altering host-plant quality

Little is known about how plant nutritional and defensive qualities interact to influence predator–prey interactions. To address this need, we provided the neo-tropical milkweed, Asclepias curassavica, with two levels of nitrogen availability and examined how altered host-plant quality influenced the responses of a specialist aphid, Aphis nerii, and a coccinellid predator, Harmonia axyridis. Aphis nerii uses A. curassavica for multiple resources, including nutrition and sequestration of cardenolides for defense against natural enemies. Increased nitrogen availability improved A. curassavica quality by decreasing carbon-to-nitrogen (C:N) ratios and cardenolide concentrations, resulting in A. nerii that also had lower C:N ratios and cardenolide concentrations. Aphis nerii population growth was higher on plants with high nitrogen availability, compared with aphids on plants with low nitrogen availability. In no-choice feeding trials, Harmonia axyridis consumed more high C:N ratio aphids, suggesting a potential compensatory response to reduced aphid nutritional quality. Additionally, H. axyridis were able to consume more low-quality aphids at the expense of increasing exposure to increased cardenolide concentrations, suggesting that interactions between H. axyridis and A. nerii may be strongly influenced by prey nutritional quality. This work highlights the need to consider how variation in plant quality influences herbivore nutritional and defensive quality when examining mechanisms that influence predator–prey interactions.     

Poisoned plusiines: toxicity of milkweed latex and cardenolides to some generalist caterpillars

Summary. Larvae of Trichoplusia ni Hübner (Lepidoptera: Noctuidae) that ingested latex from Asclepias curassavica L. (Asclepiadaceae) often regurgitated and convulsed with spasms before becoming immobile and unresponsive. Some larvae required over a day to recover sufficiently to feed. Latices from four other plant species in three families were all deterrent, but none caused detectable poisoning. The toxicity of A. curassavica latex was evidently due to cardenolides because pure cardenolides had similar effects when ingested by T. ni. Other species of noctuid caterpillars (Rachiplusia ou Guenée, Anagrapha falcifera Kirby, and Autographa precationis Guenée) sometimes also suffered spasms and temporary immobility when fed A. curassavica latex. A more distantly related noctuid, Spodoptera ornithogalli, was deterred by the latex, but showed no overt physiological responses at the dosage tested. T. ni larvae failed to develop on intact leaves of A. curassavica, on leaves with latex canals deactivated by midrib severance, and on excised leaves. Similarly, larvae reared on excised A. syriaca L. leaves to the final instar died when transferred to A. curassavica leaves with either intact or severed midribs. The final instar larvae sometimes suffered from spasms and immobility even when confined on leaves with depressurized canal systems. Evidently, cardenolides stored outside the latex system suffice to poison larvae. We conclude that cardenolides in A. curassavica have potent physiological effects on some generalist caterpillars and that the presence of these compounds both inside and outside laticifers effectively protects the plant. Received 30 June 1999; accepted 18 October 1999     

Cardenolide content,emetic potency,and thin-layer chromatography profiles of monarch butterflies,Danaus plexippus,and their larval host-plant milkweed,Asclepias humistrata,in Florida

Summary This paper is the fourth in a series on cardenolide fingerprints of monarch butterflies (Danaus plexippus, Danainae) and their host-plant milkweeds (Asclepiadaceae) in the eastern United States. Cardenolide concentrations ofAsclepias humistrata plants from north central Florida ranged from 71 to 710 µg/0.1 g dry weight, with a mean of 417 µg/0.1 g. Monarchs reared individually on these plants contained cardenolide concentrations ranging from 243 to 575 µg/0.1 g dry weight, with a mean of 385 µg/0.1 g. Cardenolide uptake by butterflies was independent of plant concentration, suggesting that sequestration saturation occurs in monarchs fed cardenolide-rich host plants. Thinlayer chromatography resolved 19 cardenolides in the plants and 15 in the butterflies. In addition to humistratin,A. humistrata plants contained several relatively non-polar cardenolides of the calotropagenin series which are metabolized to more polar derivatives in the butterflies. These produced a butterfly cardenolide fingerprint clearly distinct from those previously established for monarchs reared on otherAsclepias species. In emetic assays with the blue jay,Cyanocitta cristata, the 50% emetic dose (ED₅₀) per jay was 57.1 µg, and the average number of ED₅₀ units per butterfly was 13.8, establishing that this important south eastern milkweed produces highly emetic, chemically defended monarchs. Our data provide further support for the use of cardenolide fingerprints of wild-caught monarchs to make ecological predictions concerning defence against natural enemies, seasonal movement and larval host-plant utilization by monarch butterflies during their annual cycle of migration, breeding and overwintering.     

Molecular studies on the ouabain binding site of the Na+, K+-ATPase in milkweed butterflies

Summary. The Na⁺, K⁺-ATPase of the Monarch butterfly (Danaus plexippus) is insensitive to the inhibition by cardiac glycosides due to an amino acid replacement: histidine instead of asparagine at position 122 of the α-subunit representing the ouabain binding site. By PCR amplification of the DNA sequence of this site, a PCR product of 270 bp was obtained from DNA extracted from Danainae species (Danaus plexippus, D. chrysippus, D. gillipus, D. philene, D. genutia, Tirumala hamata, Euploea spp., Parantica weiskei, P. melusine), Sphingidae (Daphnis nerii) and mimics of milkweed butterflies (Hypolimnas missipus, Limenitis archippus and L. arthemis, Nymphalidae). Analysis of the nucleotide sequences revealed that the single point mutation in the ouabain binding domain (AAC-Asn for CAC-His) was present only in Danaus plexippus, but not in the other species investigated. Since these milkweed butterflies also store cardenolides, other structural modifications of the Na⁺, K⁺-ATPase may have occurred or other strategies of cardenolide tolerance have been developed. Received 15 May 2000; accepted 29 June 2000     

Milkweeds,monarch butterflies and the ecological significance of cardenolides

Summary The contribution of Miriam Rothschild to the monarch cardenolide story is reviewed in the light of the 1914 challenge by the evolutionary biologist, E.B. Poulton for North American chemists to explain the chemical basis of unpalatability in monarch butterflies and their milkweed host plants. This challenge had lain unaccepted for nearly 50 years until Miriam Rothschild took up the gauntlet and showed with the help of many able colleagues that monarchs are aposematically coloured because they sequester toxic cardenolides from milkweed host plants for use as a defence against predators. By virtue of Dr Rothschild's inspiration and industry, and subsequently that of Lincoln Brower and his colleagues, this tritrophic interaction has become a familiar paradigm for the evolution of chemical defences and warning colouration. We now know that the cardenolide contents of different milkweeds vary quantitatively, qualitatively and spatially, both within and among species and we are starting to appreciate the implications of such variation. However, as Dr Rothschild has pointed out in her publications, cardenolides have sometimes blinded us to reality and it is curious how little evidence there is for a defensive function to cardenolides in plants — especially against adapted specialists such as the monarch. Thus the review will conclude with a discussion of the significance of temporal variation and induction of cardenolide production in plants, the lethal plant defence paradox and an emphasis on the dynamics of the cardenolide-mediated interaction between milkweeds and monarch larvae.     

Leaf choices of nest-building crab spiders (Misumena vatia)

Summary Crab spiders (Misumena vatia, Thomisidae) selected leaves of common milkweed (Asclepias syriaca) over those of spreading dogbane (Apocynum androsaemifolium), pasture rose (Rosa carolina), and chokecherry (Prunus virginiana) as nest-sites in pairwise experiments among these four substrates. Nesting success was higher on milkweed than on the other three plant species. Early survival of the clutches was thus correlated with the preferences exhibited by their mothers. The major source of nest failure was parasitization by the ichneumonid wasp, Trychosis cyperia, which accounted for over 50 per cent of the nest losses.     

Glues or poisons: which triggers vein cutting by monarch caterpillars?

Summary. Late instar larvae of the monarch butterfly (Danaus plexippus L., Lepidoptera: Nymphalidae) deactivate the latex defense of milkweeds by chewing a furrow in the leaf midrib or petiole. The larva then feeds beyond the cut where latex outflow is minimal. If a larva does encounter latex exudate during feeding, it often returns to its initial cut to damage the midrib or petiole more extensively before resuming feeding. We used this response to latex as an assay for testing what cue triggers vein cutting. A sticky solution of polyethylene glycol and a mixture of cardenolides both were ineffective; drops applied to the mouthparts of feeding monarchs failed to elicit renewed vein cutting. Activity resided instead within a methanol extract of the supernatant obtained from centrifuged latex of the milkweed, Asclepias curassavica L. (Asclepiadaceae). Treatment with proteinase K did not eliminate activity documenting that the active compound is unlikely to be proteinaceous. Our results indicate that latex adhesiveness and low polarity cardenolides are relatively unimportant releasers of vein cutting. We propose that milkweeds contain noncardenolide stimulants of vein cutting; these compounds presumably serve a defensive role for milkweeds. Over 50 species of insects are known to employ vein-cutting before feeding on plants with canal-borne exudates; most of these species are dietary specialists like the monarch. Our behavioral assay provides a novel approach for identifying ecologically-significant compounds in the exudates of their diverse host plants.     

Target-site sensitivity in a specialized herbivore towards major toxic compounds of its host plant: the Na+K+-ATPase of the oleander hawk moth (Daphnis nerii) is highly susceptible to cardenolides

The caterpillars of the oleander hawk moth, Daphnis nerii (Linnaeus, 1758) (Lepidoptera: Sphingidae) feed primarily on oleander (Nerium oleander). This plant is rich in cardenolides, which specifically inhibit the Na⁺K⁺-ATPase. Since some insects feeding on cardenolide plants possess cardenolide-resistant Na⁺K⁺-ATPases, we tested whether D. nerii also possesses this strategy for circumventing cardenolide toxicity. To do so, we established a physiological assay, which allowed direct measurement of Na⁺K⁺-ATPase cardenolide sensitivity. Using Schistocerca gregaria, as a cardenolide-sensitive reference species, we showed that D. nerii Na⁺K⁺-ATPase was extremely sensitive to the cardenolide ouabain. Surprisingly, its sensitivity is even higher than that of the cardenolide-sensitive generalist, S. gregaria. The presence or absence of cardenolides in the diet of D. nerii did not influence the enzyme’s cardenolide sensitivity, indicating that target-site insensitivity is not inducible in this species. However, despite the sensitivity of their Na⁺K⁺-ATPase, caterpillars of D. nerii quickly recovered from an injection of an excessive amount of ouabain into their haemocoel. We conclude that D. nerii possesses adaptations, which enable it to feed on a cardenolide-rich diet other than that previously described in cardenolide specialized insects, and discuss other potential resistance mechanisms.     

Demographic Effects of Harvesting Epiphytic Bromeliads and an Alternative Approach to Collection

Abstract: Hundreds of epiphytic bromeliads species are harvested from the wild for trade and for cultural uses, but little is known about the effects of this harvest. We assessed the potential demographic effects of harvesting from the wild on 2 epiphytic bromeliads: Tillandsia macdougallii, an atmospheric bromeliad (adsorbs water and nutrients directly from the atmosphere), and T. violaceae, a tank bromeliad (accumulates water and organic material between its leaves). We also examined an alternative to harvesting bromeliads from trees—the collection of fallen bromeliads from the forest floor. We censused populations of T. macdougallii each year from 2005 to 2010 and of T. violaceae from 2005 to 2008, in Oaxaca, Mexico. We also measured monthly fall rates of bromeliads over 1 year and monitored the survival of fallen bromeliads on the forest floor. The tank bromeliad had significantly higher rates of survival, reproduction, and stochastic population growth rates (λ_s) than the atmospheric bromeliad, but λ_s for both species were <1, which suggests that the populations will decline even without harvest. Elasticity patterns differed between species, but in both, survival of large individuals had high elasticity values. No fallen bromeliads survived more than 1.5 years on the forest floor and the rate of bromeliad fall was comparable to current harvest rates. Low rates of population growth recorded for the species we studied and other epiphytic bromeliads and high elasticity values for the vital rates that were most affected by harvest suggest that commercial harvesting in the wild of these species is not sustainable. We propose the collection of fallen bromeliads as an ecologically and, potentially, economically viable alternative.     

Leaf reddening in the seagrass Thalassia testudinum in relation to anthocyanins, seagrass physiology and morphology, and plant protection

Numerous seagrass species growing in high-light environments develop red coloration in otherwise green leaves, yet the ecophysiology of leaf reddening in seagrasses is poorly understood. To increase our understanding of the process of leaf reddening in Thalassia testudinum found in the lower Florida Keys (USA), we identified the molecules responsible for red coloration in leaves and compared physiological, morphological, and growth attributes of entirely red-leafed shoots to entirely green-leafed shoots. We determined that four anthocyanin molecules are responsible for red coloration in leaves. In addition, we found that red leaves had higher concentrations of photoprotective pigments (anthocyanins and UV-absorbing compounds), higher effective quantum yields (ΔF/F _m′) at midday, and were shorter, narrower, and weighed less than green leaves. No significant difference in growth rates was observed between red- and green-leafed shoots, but patches of red-leafed shoots had shorter canopy heights and smaller LAI compared to patches of green-leafed shoots. Our results demonstrate that leaf reddening in T. testudinum is caused by high concentrations of anthocyanins, is associated with physiological and morphological attributes, and acts as a sunscreen since red leaves were able to maintain high effective quantum yields at high light intensities.     

Effects of temperature on yolk utilization,initial growth,and behaviour of unfed marine fish-larvae

The effect of temperature on yolk utilization, initial growth and behaviour of larvae of four species of marine fishes, i.e.,Acanthopagrus schlegeli, Engraulis japonica, Pagrus major andParalichthys olivaceus, was investigated under laboratory conditions at Hiroshima in 1989. The yolk sac was absorbed earlier with increasing temperature for all species. Morphological characters such as pectoral fin appearance, mouth opening and eye pigmentation differed from species to species over the range of experimental temperatures, and the sequence of development of these characters varied with temperature even within the same species. Temperature did not have a large influence on the maximum growth of unfed larvae, but had a clear effect on the time to starvation, occurring earlier at higher temperatures. Larval behaviour as indicated by time spent moving was also influenced by temperature; behavioural activity was greater at higher temperatures for all species examined. The effect of temperature on the early stage of larval life is discussed in terms of behavioural and morphological developments, as well as on yolk utilization, and its influence on larval survival in nature and under rearing conditions is evaluated.     

A response-surface approach to the combined effects of temperature and salinity on the larval development of Adula californiensis (Pelecypoda: Mytilidae). I. Survival and growth of three and fifteen-day old larvae

Laboratory experiments of a factorial design were used to examine the combined effects of temperature and salinity on the survival and growth of early and late-stage larvae of Adula californiensis (Phillippi, 1847). Response-surface curves were generated to predict optimal conditions for survival and growth in order to better understand the successful recruitment of this species within the Yaquina Bay estuary (Oregon, USA). Three-day old cultured larvae were more sensitive to reduced salinity than were 15-day old larvae. However, the 15-day old larvae showed a narrower temperature tolerance than the 3-day old larvae. A. californiensis larvae survived over a wider range of temperatures near optimum salinities than at salinities near their lower tolerance limit, and conversely. Temperature and salinity ranges for maximum survival (10° to 15°C, 31 to 33) were narrower than the ranges which occur within the estuary where the adult populations exist. Larval size did not increase markedly during the 15-day rearing period, and was not greatly affected by temperature or salinity. No statistically significant temperature-salinity interaction was found for either survival or growth.     

Laboratory study of predation by Aurelia aurita on larvae of cod,flounder, plaice and herring: development and vulnerability to capture

Capture success of the medusa Aurelia aurita preying on various developmental stages of fish larvae was measured together with larval reactivity and escape speed after being stung. These experiments were conducted in the spring of 1983 with A. aurita medusae collected from Loch Etive, Scotland and laboratory-reared larvae of Gadus morhua L., Platichthys flesus L., Pleuronectes platessa L. and Clupea harengus L. Capture success of the medusae increased with medusa size, but decreased with advancing larval development. Smaller species of larvae were more vulnerable to capture. Larval reactivity to encounters with medusae increased with advancing development, and larger species of larvae were more reactive to encounters. Larval escape swimming speeds also increased with advancing larval development and size. These results indicate that earlier stages of larvae within a species and smaller species of larvae at a given stage are more vulnerable to predation by medusae since they are less reactive to encounters. Apparently they are more susceptible to the effects of neurotoxins. Predation rates on different developmental stages of herring larvae are documented and compared with rates predicted by a predation model. Predictions fell within the range of observed predation rates, but tended to overestimate rates by larger medusae feeding on larger herring larvae. This indicates the possibility of predator satiation and/or behavioural avoidance.     

Survival, growth and reproduction of the salt-marsh amphipod Uhlorchestia spartinophila reared on natural diets of senescent and dead Spartina alterniflora leaves

The talitrid amphipod Uhlorchestia spartinophila lives in close association with standing-dead leaves of the smooth cordgrass Spartina alterniflora Loisel in salt marshes along the Atlantic coast of North America. This study probed the strength of the trophic link between the amphipod population and the decomposition process in this detrital-based ecosystem. We measured survival, growth and reproductive output in groups of amphipods reared for 6 wk on five diets derived from sheath and blade portions of S. alterniflora leaves just prior to (senescent) and during (dead) decomposition. In unfed treatments, the daily specific mortality rate was 0.391 and starved amphipods survived no longer than 11 d. Among the fed treatments, a diet of senescent sheaths resulted in the lowest survival (20%) and yielded no offspring. Groups fed senescent blades, dead sheaths, dead blades and unwashed dead sheaths had survival rates of 56 to 84% and produced 5.0 to 12.5 offspring replicate⁻¹. Sex ratio usually favored females, but approached unity in treatments with high overall survival, suggesting that quality of available food resources may influence sex ratio in this species. Mean specific growth rates (mm mm⁻¹ d⁻¹) ranged from 0.013 to 0.016, and matched previous estimates of growth from field populations. Overall ecological performance (survival + growth + reproduction) was similar for all food treatments, except senescent sheaths, which yielded a final mean (±SD) dry biomass (0.4 ± 0.42 mg replicate⁻¹) of amphipods significantly lower than that of other diets (1.7 ± 0.81 to 2.6 ± 0.69 mg replicate⁻¹). Natural diets derived from decomposing cordgrass leaves can fulfill the nutritional requirements of U. spartinophila populations, but variation in initial amounts of living fungal biomass among the five experimental diets only partially explained the responses of amphipods in our experiment. Structural characteristics and variation in rates of fungal occupation within different portions of cordgrass leaves may affect the amphipod's ability to access plant production made available by decomposers. Received: 12 December 1996 / Accepted: 18 December 1996     

Robustness in life history of the brown seaweed Ascophyllum nodosum (Fucales, Phaeophyceae) across large scales: effects of spatially and temporally induced variability on population growth

Understanding the demography and function of biotope-forming seaweed species is of great importance for the conservation of the target species itself, as well as its associated organisms. The brown seaweed Ascophyllum nodosum is fundamental for the functioning of coastal marine ecosystems in the North Atlantic. In this study, we use a data-based size-classified matrix model to investigate the temporal and spatial variability in demography, and the environment-specific stochastic sensitivity and elasticity, of two A. nodosum populations, one in western Sweden and one on the Isle of Man in the Irish Sea. A significant difference between the two populations was that the Swedish population had comparably low and more variable stochastic population growth rate (λ _s). This pattern was partly explained by the relatively high and varying mortality rates during extreme ice-years in Sweden, and by the lower survival of small individuals during all years. There were also fewer large individuals in Sweden due to lower transitions to the larger size-classes and higher probability of shrinkage. Sensitivities were analogous in the two populations, and showed a high selection pressure for increased individual growth. Elasticities were also similar, with the exception that survival of the smallest individuals (i.e., transition a _1,1), had a higher elasticity on the Isle of Man. Overall, the stochastic growth rate (λ _s) was most sensitive to proportional changes in loop- (i.e., survival within size-class) and, to some extent, growth-transitions in both study areas. These results show that structurally and demographically diverging A. nodosum populations may be similarly sensitive to changes in vital rates. This, in turn, indicates a plastic life history of A. nodosum that may cope with large environmental variability. The results further suggest that environmental change affecting the survival or growth of the larger, reproductive A. nodosum individuals could have severe and regional effects on the abundance and biomass of this species, with potential negative effects on the biodiversity of the associated communities.     

Individual effects and interactions of salinity,temperature, and zinc on larval development of the grass shrimp Palaemonetes pugio. I. Survival and developmental duration through metamorphosis

Larvae of the estuarine grass shrimp Palaemonetes pugio (Holthuis) were reared from hatch through successful completion of metamorphosis in 80 combinations of salinity (3 to 31%), temperature (20° to 35°C), and zinc (0.00 to 1.00 ppm Zn⁺⁺). Response-surface methodology was employed to depict the individual effects and interactions of the three factors on survival and developmental duration through total larval development. Outside the optimal salinity-temperature conditions of 17 to 27 S and 20° to 27°C, viability of larvae was reduced by both the individual effects of salinity and temperature and interactions between the two factors. Survival capacity of larvae and resistance adaptations to salinity and temperature were progresively reduced by zinc concentrations from 0.25 to 1.00 ppm Zn⁺⁺. Response-surface analysis of the data suggested that the duration of total larval development of P. pugio was least at salinities from 18 to 23 and at temperatures from 30° to 32°C. At both higher and lower salinity-temperature conditions and in increasing zinc concentrations from 0.25 to 1.00 ppm Zn⁺⁺, developmental rates were retarded. A significant zinc-temperature interaction existed, whereby increasing zinc concentrations reduced both survival and developmental rates of larvae more at suboptimal temperatures. Larval resistance to zinc toxicity was least at supraoptimal salinities, indicative of a significant zinc-salinity interaction. The reduced viability, restricted euryplasticity, and retarded developmental rates of P. pugio larvae developing in media with low-level zinc contamination would limit the distributive properties of the pelagic phase in the life cycle of the species and reduce recruitment both into and out of the parent estuarine population.     

Mechanisms of habitat segregation between an invasive (Mytilus galloprovincialis) and an indigenous (Perna perna) mussel: adult growth and mortality

The invasive mussel Mytilus galloprovincialis and the indigenous mussel Perna perna coexist intertidally on the south coast of South Africa through partial vertical habitat segregation: M. galloprovincialis dominates the upper shore and P. perna the lower shore. Recruitment patterns can explain the zonation of P. perna, but not the invasive species. We examined the role of post-recruitment interactions by measuring spatial and temporal differences in adult growth and mortality rates of the two species. Specifically, we tested the hypothesis that interspecific differences in growth and mortality reflect adult distribution patterns. The two study locations, Plettenberg Bay and Tsitsikamma, are 70 km apart with two sites (separated by 300–400 m) per location, each divided into three vertical zones. Growth was measured seasonally using different marking methods in 2001 and 2003. Cumulative adult mortality was measured through summer in 2003/2004. Both species generally grew more slowly upshore, but they showed different effects of season. For P. perna, growth was significantly reduced in winter in the low zone, but unaffected by season in the high zone. For M. galloprovincialis, growth was either unaffected by season or increased in winter, even in the high zone. Thus, growth of P. perna and M. galloprovincialis was reduced under cool winter and warm summer temperatures, respectively; and while growth was more similar between species in summer, M. galloprovincialis grew much faster than P. perna in winter. Mortality of P. perna increased upshore. For M. galloprovincialis, mortality was not zone-dependent and was significantly greater than for P. perna on the low-shore and (generally) across the shore in Tsitsikamma. Both species had higher growth and mortality rates in Plettenberg Bay than in Tsitsikamma. Thus, P. perna seems able to maintain spatial dominance on the low-shore and at certain sites because of higher mortality of M. galloprovincialis. We conclude that seasonality in growth of the two species reflects their biogeographic affinities and that coexistence is possible through pre-recruitment effects that limit the vertical distribution of P. perna and post-recruitment effects that limit M. galloprovincialis.     

Essential fatty acid (docosahexaenoic acid, DHA) availability affects growth of larval herring in the field

Larval fish growth and survival depends not only on prey quantity, but also on prey quality. To investigate effects of prey fatty acid concentration on larval herring growth, we collected different prey organisms and larval herring (Clupea harengus L.) in the Kiel Canal during the spring season of 2009. Along with biotic background data, we analysed fatty acids both in prey organisms and in the larvae and used biochemically derived growth rates of the larvae as the response variable. Larval herring reached their highest RNA/DNA derived growth rates only at high docosahexaenoic acid (DHA) concentration. When the ratio of copepodids to lesser quality cirriped nauplii was low, larval growth and larval DHA concentration were both significantly negatively affected. This was true even as prey abundance was increasing. This finding indicates that even in mixed, natural feeding conditions, growth variations are associated with DHA availability in larval fish.