Cytoenzymatic investigation of intracellular digestion in the symbiont-bearing hydrothermal bivalve <Emphasis Type="Italic">Bathymodiolus azoricus</Emphasis>

Invertebrates harbouring endosymbiotic chemoautotrophic bacteria are widely distributed in a variety of reducing marine habitats, including deep-sea hydrothermal vents. Bathymodiolids are dominants of the biomass at geochemically distinct vent sites of the Mid Atlantic Ridge (MAR) and thus are good candidates to study biological processes in response to site-specific conditions. To satisfy their nutritional requirements, these organisms depend to varying extent on two types of chemoautotrophic symbionts and on filterfeeding. The quantitative relationships of the nutritional modes are poorly understood. Using enzyme cytochemistry, electron microscopy and X-ray microanalysis, the structural and functional aspects of the cellular equipment necessary for lysosomal digestion was studied. We provide evidence for the following: (1) the basis of intracellular digestion of symbionts in Bathymodiolus azoricus from two geochemically distinct vent sites was not mainly in the large lysosomal bodies as previously thought (based on the membranous content resembling bacteria); (2) senescent bacteria are autolysed, possibly by bacterial acid phosphatase, that is more likely a cell cycling of the symbionts rather than an active lysosomal digestion by the host; (3) the consistent absence of hydrolases may indicate the improper use of the name “lysosome” for large vesicles at the base of the gill bacteriocytes (4) nutrient transfer in B. azoricus, therefore, may more likely be accomplished through leaking of metabolites from the symbiont to the host, not excluding lysosomal resorption of dead bacteria as an auxiliary strategy for organic molecule transfer; (5) evidence is provided for microvillar transfer of substances from the seawater that may indicate filter-feeding, in non-symbiotic ciliated gill cells of mussels from Lucky Strike; (6) two types of lysosomal vesicles can be distinguished in digestive cells based on their enzymatic content and their elemental composition.     

Nutritional exchange in a tropical tripartite symbiosis: direct evidence for the transfer of nutrients from anemonefish to host anemone and zooxanthellae

The relationship between anemones and anemonefishes is an oft-cited and endearing example of a mutualistic symbiosis. Current research on mutualistic symbioses suggests these relationships are more commonplace and have greater importance at the ecosystem level on nutrient dynamics and evolutionary processes than previously thought. Using stable isotopes ¹⁵N and ¹³C, both field and laboratory experiments were designed to investigate whether nutrient transfer from two species of resident anemonefishes (Amphiprion perideraion and A. clarkii) to host anemones (Heteractis crispa) occurs. Mass spectroscopy indicated that both ¹⁵N and ¹³C were significantly elevated in the tissues of anemonefishes and in both host anemone and zooxanthellae fractions. These experiments provide the first direct empirical evidence of nitrogen and carbon transfer from resident anemonefishes to host anemones and endosymbiotic zooxanthellae. Such transfer of elements within this intriguing tripartite association underscores the central role that nutrient dynamics contributes to the evolutionary processes of these marine symbioses.     

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.     

Bearded vultures (<Emphasis Type="Italic">Gypaetus barbatus</Emphasis>) prefer fatty bones

In animal species, prey processing and the provisioning of nutrients are subject to several constraints related with finding, ingesting and processing food. In most bird species, these constraints are obvious as a consequence of food morphology. In the case of the bearded vulture (Gypaetus barbatus), in comparison with other species, its behavioural and physiological adaptations apparently allow this vulture to ingest bone remains irrespective of their morphology. Here, by comparing bones delivered to the nest to be consumed (selected) and remains found at an experimental feeding station and at bone-breaking sites or ossuaries (rejected), I tested whether bearded vultures are capable of choosing from among the various anatomical parts of an animal carcass in relation to their fatty acid content (nutrient concentration hypothesis), their size (width-reduction hypothesis) or both. The results suggest that bearded vultures prefer the fatty anatomical parts (with a high percentage of oleic acid) of an animal carcass regardless of bone length, although bone morphology as a consequence of handling efficiency or the ingestion process may also play a secondary role in food selection. The close association between the bones selected and their high fat value implies an optimisation of foraging time and of the increased energy gained from the food. This is in line with selective foraging to redress specific nutritional imbalances (nutrient concentration hypothesis) and, secondarily, the width-reduction hypothesis.     

Testing the Stochastic Dynamic Methodology (StDM) as a management tool in a shallow temperate estuary of south Europe (Mondego, Portugal)

A long-term monitoring program has been carried out since the early 1990s in the Mondego estuary, on Portugal's west coast, which is presently under heavy human pressure. In this shallow warm-temperate estuary, a significant macroalgal proliferation has been observed, which is a clear sign of nutrient enrichment. As a result of competition with algae, the extension of the seagrass meadows (mainly Zostera noltii) has been reduced. The present paper examined the applicability of a holistic Stochastic Dynamic Methodology (StDM) in predicting the tendencies of trophic key-components (macrophytes, macroalgae, benthic macroinvertebrate and wading birds) as a response to the changes in estuarine environmental conditions. The StDM is a sequential modelling process developed in order to predict the ecological status of changed ecosystems, from which management strategies can be designed. The data used in the dynamic model construction included true gradients of environmental changes and was sampled from January 1993 to September 1995 and from December 1998 to December 2005. The dynamic model developed was preceded by a conventional multivariate statistical procedure performed to discriminate the significant relationships between the selected ecological components. The model validation was based on independent data collected from January 1996 to January 1997 and from February 1999 to April 2000 for all the state variables considered. Overall, the simulation results are encouraging since they seem to demonstrate the StDM reliability in capturing the trophic dynamics of the studied estuary, by predicting the behavioural pattern for the most part of the components selected, with a focus on the Zostera noltii meadows recovery after the implementation of important management measures.     

A Functional Approach to the Assessment of the Nutritional Value of Particulate Organic Matter

Organic matter lability can be determined with different approaches, based on the quantitative and qualitative estimate of its nutritional value. the C/N ratio and the particulate organic matter (as the sum of protein, carbohydrate and lipid bulks) vs total suspended matter (POM/TSM) deal with a quantitative assessment of the organic matter bulk, while other indices such as proteins/carbohydrates ratio (PRT/CHO) may give also qualitative information. Gordon (1970) proposed a functional approach, based on organic carbon decrease after enzymatic digestion. Coupling Gordon's approach with the evaluation of the main biochemical components (proteins, carbohydrates and lipids) contained in the particulate organic matter, a simple method to assess lability and nutritional value of organic substrates is proposed.

Water samples were collected at the surface and close to the bottom in a shallow coastal station (10 m depth) in the Marconi Gulf (Ligurian Sea, NW Mediterranean). the difference between “total labile” and “hydrolyzed labile” organic matter (respectively before and after enzymatic digestion) suggests a relevant presence of resuspended refractory materials vs about 60% of labile hydrolyzed organic matter, thus confirming the importance of a more qualitative approach to organic matter studies.     

Estimating paternity with spatial behaviour and DNA fingerprinting in the striped plateau lizard, Sceloporus virgatus (Phrynosomatidae)

The association between spatial proximity and paternity was studied in a population of the striped plateau lizard, Sceloporus virgatus. The relationship between estimated mating success and male phenotypic traits was examined for a sample of 55 males. DNA samples were obtained from 13 female-offspring families. The males with the closest spatial proximity to each female were tested as possible sires within each family. Fingerprinting with two multilocus hypervariable minisatellite probes revealed a strong correspondence between male-female spatial proximity and actual paternity. Paternity could be assigned for 72 of the 100 hatchlings. Most hatchlings with identifiable sires were attributed to a male with the highest category of spatial proximity to the mother. However, there was a low to moderate level of multiple paternity within clutches, and for some clutches probable sires could not be identified even though the most likely behavioural candidates were tested. Thus, nonterritorial males or other males lacking strong social and spatial relationships with females may achieve some degree of reproductive success. Analysis of mating success revealed that male success increased with body size, up to a point beyond which larger size conferred no advantage. Received: 7 January 1997 / Accepted after revision: 16 June 1997     

Effects of nutritional history on nitrogen assimilation in congeneric temperate and tropical scleractinian corals

The nutritional history of corals is known to affect metabolic processes such as inorganic nutrient uptake and photosynthesis, but little is known about how it affects assimilation efficiency of ingested prey items or the partitioning of prey nitrogen between the host and symbiont. The temperate scleractinian coral Oculina arbuscula and its tropical congener Oculina diffusa were acclimated to three nutritional regimes (fed twice weekly, starved, starved with an inorganic nutrient supplement), then fed Artemia nauplii labeled with the stable isotope tracer ¹⁵N. Fed corals of both species had the lowest assimilation efficiencies (36–51% for O. arbuscula, 38–57% for O. diffusa), but were not statistically different from the other nutritional regimes. Fed and starved corals also had similar NH₄⁺ excretion rates. This is inconsistent with decreased nitrogen excretion and reduced amino acid catabolism predicted by both the nitrogen recycling and conservation paradigms. In coral host tissue, ~90% of the ingested ¹⁵N was in the TCA-insoluble (protein and nucleic acids) and ethanol-soluble (amino acids/low molecular weight compounds) within 4 h of feeding. The TCA-insoluble pool was also the dominant repository of the label in zooxanthellae of both species (40–53% in O. arbuscula, 50–60% in O. diffusa). However, nutritional history had no effect on the distribution of prey ¹⁵N within the biochemical pools of the host or the zooxanthellae for either species. This result is consistent with the nitrogen conservation hypothesis, as preferential carbon metabolism would minimize the effects of starvation on nitrogen-containing biochemical pools.Communicated by P.W. Sammarco, Chauvin     

Effects of within- and among-patch experiences on the patch-leaving decision rules in an insect parasitoid

The present study aimed to address how an insect parasitoid makes patch-departure decisions from various types of host patches and how previous patch experiences in the environment modify this decision-making process. Experiments were done with the parasitic wasp Aphidius rhopalosiphi attacking the grain aphid Sitobion avenae. In the experiments, wasps were observed in a laboratory environment containing several patches of various host densities, and behavioural records were analysed using a Coxs proportional hazards model. Consideration of the effect of the within-patch experience gave a classic pattern of patch-leaving decision rules in parasitoids: A. rhopalosiphi used local information on host quality (i.e. numbers of ovipositions or rejections) and availability (i.e. patch density) to determine departure decision. However, consideration of previous patch experiences provided evidence that these departure rules are fundamentally dynamic, responding to the physiological state of the animal and the information it has about its environment. Results showed that A. rhopalosiphi decreased its tendency to leave the visited patch after an oviposition. However, when a female has already laid several other eggs in the environment, such an incremental mechanism gradually switched to a decremental one. Hence, A. rhopalosiphi responded to egg-load depletion by leaving the visited patches sooner and by depositing a smaller number of eggs in those patches, which probably led to a decreased level of superparasitism. Results also indicated that previous experiences enabled wasps to estimate spatial host distribution and then to adjust their behavioural decisions accordingly. Thus, A. rhopalosiphi was shown to adjust its patch residence time according to the quality and the number of the patches previously visited. These proximate mechanistic rules adopted by A. rhopalosiphi females are discussed in the context of general predictions from optimality models.Communicated by D. Gwynne     

Effects of nutrient concentration and litter cover on quantitative shoot parameters and belowground biomass of Zizania latifolia (L)

This study investigated the effect of nutrient concentration and litter cover on the development of quantitative shoot parameters and belowground biomass (BGB) production of Zizania latifolia. Zizania latifolia is a common emergent aquatic species in East Asia. Four treatments were done at the study site, and were observed between May 2003 and December 2005. The treatments are namely, high nutrient (HN) with litter cover (HNWL), high nutrient without litter cover (HNNL), low nutrient (LN) with litter cover (LNWL), and low nutrient without litter cover (LNNL). The quantitative shoot parameters and BGB had higher values for treatments with high nutrient (HN) compared to the low nutrient treatments (LN), independent of the presence of litter cover. Furthermore, the life span of the secondary shoots was also higher in HN treatments compared to LN treatments. The BGB productivity was higher in the HNNL treatment compared to the other treatments. The LNWL treatment showed the least developed quantitative shoot parameters, e.g. plant height, and the lowest BGB for Z. latifolia. It was generally observed that the combined effects of low nutrients and litter cover negatively affected shoot development and BGB production.     

Fluctuating asymmetry and sperm transfer in male decorated field crickets (Gryllodes sigillatus)

Fluctuating asymmetry (FA) in limb size in female decorated field crickets (Gryllodes sigillatus) was associated with a reduction in the size of the spermatophore and the amount of sperm transferred by males and an increase in the time taken to transfer a spermatophore following introduction of a female. There was a weaker negative relationship between limb asymmetry in males and sperm number but no significant relationship between asymmetry in either sex and spermatophylax size. In line with a previous study, female size did not appear to influence spermatophore production or mating decisions by males. The results imply that developmental instability affects both gamete production and mating decisions among males, although the relationships between spermatophore size, sperm number and asymmetry in females are unlikely to be the result of males perceiving differences in female FA. Received: 24 July 1999 / Received in revised form: 22 November 1999 / Accepted: 31 December 1999     

Female preferences based on male nutritional chemical traits

Selection favors females that attend to reliable information about male genetic quality and fitness. Male nutritional condition can be a significant sign of mate quality since poor nutrition can be related to reduced sperm quality, low sperm quantity, sexually transmitted diseases, and parasites. We tested whether female Nile tilapia, Oreochromis niloticus, preferred the chemical cues of males that were well fed with high-protein diet over those fed with low-protein diet. Females do not only discriminate between males but also show a preference for well-fed males, discriminating between the odors with respect to nutritional state, suggesting that they were responding to a food-specific chemical cue. It is therefore likely that nutritional condition is related to the production of pheromones in males. Our results suggest that information about male nutritional state can be conveyed in chemical cues and that females attend to these cues during mate choice.     

Cysteine-glutathione disulfide, the sperm-release pheromone of the marine polychaete Nereis succinea (Annelida: Polychaeta)

Summary. In the marine polychaete Nereis succinea (Frey & Leuckart 1847) a sex pheromone was isolated from the coelomic fluid of sexually mature females and identified by NMR studies and independent synthesis. This pheromone is released by the females during reproduction together with eggs and coelomic fluid into the free water column and induces sperm release of surrounding males. Its structure was ascertained as L-cysteine-glutathione disulfide. It exhibited a response threshold of 0.6 · 10⁻⁷ M. Received 15 July 1997; accepted 25 November 1997.     

Improving human micronutrient nutrition through biofortification in the soil–plant system: China as a case study

Micronutrient malnutrition is a major health problem in China. According to a national nutritional survey, approximately 24% of all Chinese children suffer from a serious deficiency of iron (Fe) (anemia), while over 50% show a sub-clinical level of zinc (Zn) deficiency. More than 374 million people in China suffer from goiter disease, which is related to iodine (I) deficiency, and approximately 20% of the Chinese population are affected by selenium (Se) deficiency. Micronutrient malnutrition in humans is derived from deficiencies of these elements in soils and foods. In China, approximately 40% of the total land area is deficient in Fe and Zn. Keshan and Kaschin-Beck diseases always appear in regions where the soil content of Se in low. The soil–plant system is instrumental to human nutrition and forms the basis of the “food chain” in which there is micronutrient cycling, resulting in an ecologically sound and sustainable flow of micronutrients. Soil-plant system strategies that have been adopted to improve human micronutrient nutrition mainly include: (1) exploiting micronutrient-dense crop genotypes by studying the physiology and genetics of micronutrient flow from soils to the edible parts of crops; (2) improving micronutrient bioavailability through a better knowledge of the mechanisms of the enhancers’ production and accumulation in edible parts and its regulation through soil-plant system; (3) improving our knowledge of the relationship between the content and bioavailability of micronutrients in soils and those in edible crop products for better human nutrition; (4) developing special micronutrient fertilizers and integrated nutrient management technologies for increasing both the density of the micronutrients in the edible parts of plants and their bioavailability to humans.     

Incubation temperature has long-term effects on behaviour of young Pine snakes (Pituophis melanoleucus)

Summary Eggs of pine snakes (Pituophis melanoleucus) were incubated at constant temperatures of 21°, 23°, 26°, 28°, 30° and 32° C to determine behavioural differences as a function of incubation temperature. For all behavioural and physiological measures hatchlings from medium incubation temperatures (26°, 28°) performed tests better than those hatchlings from eggs incubated at low temperatures (21°, 23°). For some behavioural tests hatchlings from eggs incubated at high temperature (30°, 32°) performed less well than medium temperature hatchlings. These differences were not due to ambient temperatures or age of testing since these were held constant. Some of the behavioural differences persisted for 24 weeks.     

Allocation of herbivory-induced hydroxamic acids in the wild wheat Triticum uniaristatum

Summary. We characterized the induction of hydroxamic acids (Hx) by aphid infestation in the wild wheat Triticum uniaristatum by addressing the following questions: i) Do different leaves have similar responses to aphid damage?, ii) Is the Hx induction localized or systemic?, iii) How long does the induction last?, and iv) Is the degree of damage related to the magnitude of induced Hx? Based on earlier results on this wheat/aphid system (lack of costs of Hx induction) we expected to find the plant exhibiting cost-saving patterns of response to herbivory. Aphid infestation in the primary leaf led to induced levels of Hx, but no differences in Hx levels were found after infestation of the secondary leaf. Induction of Hx was restricted to the infested leaf (primary leaf). Induced Hx levels exhibited by the primary leaf at the end of aphid infestation were not observed 2 days later. Finally, different aphid densities (between 10 and 40 aphids per leaf) did not produce significant differences in Hx levels in infested primary leaves. Characteristics of Hx induction by aphid infestation in T. uniaristatum partially support the expected cost-saving patterns in the allocation of induced defenses. Received 15 January 1997; accepted 7 July 1997.     

Gpi genotypic effect on quantitative traits in the northern bay scallop,Argopecten irradians irradians

The relationships between glucose-6-phosphate isomerase (Gpi) genotype and quantitative variation related to reproduction and growth were explored over the lifespan of a single cohort of northern bay scallops, Argopecten irradians irradians (Lamarck), from September 1986 to April 1988 in the Niantic River estuary, Connecticut, USA. Analyses revealed that Gpi genotype explained an increasingly significant proportion of variation in scallop size (up to 15%) as the cohort aged. The genotype-specific effects were consistent across sampling dates and among measured traits. There was no evidence for heterosis with respect to size at this locus; however, rare genotypes contributed substantially to the relationship and showed some tendency to fall on the extremes of the phenotypic distribution. The strength and consistency of the genotypic effect on scallop size suggest that genetic variation for Gpi, or some locus in linkage disequilibrium with Gpi, may translate into biochemical and/or physiological variation and affect fitness in this species.     

Inter-individual variation in sequestration (as measured by energy dispersive spectroscopy) predicts efficacy of defensive secretion in lubber grasshoppers

Summary. We tested whether inter-individual variation in concentration of sequestered compounds predicts defensive efficacy of the defensive secretion of onion-fed Eastern lubber grasshoppers. When fed onion, lubbers have the ability to sequester sulfur compounds into their defensive secretion, and the secretion from onion-fed lubbers is more deterrent of ants than the secretion from lubbers fed other diets (Jones et al. 1989). To test three hypotheses, we developed a method for measuring total sequestered sulfur in the secretion using energy dispersive spectroscopy (EDS). First, we found that lubbers fed an ad libitum, monophagous diet of onion produce more secretion and have higher concentrations of total sulfur in their secretions than lubbers fed other diets. These sulfur concentrations varied three-fold; we believe that this variation in the composition of defensive secretions stems from physiological variation. Second, our method was sensitive enough to detect sulfur in the secretion of some lubbers that had been fed onion only once. Third, the sulfur concentration of a secretion sample significantly predicted its ability to deter ants from foraging. Hence, our results suggest that inter-individual variation in defensive chemistry in insects offered the same diet may be ecologically important. Received 25 November 1997; accepted 5 February 1998.     

Differences in reproductive strategies of morphs of the genus Echinaster (Echinodermata: Asteroidea) from the Eastern Gulf of Mexico

Two morphs of the asteroid genus Echinaster, E. Type 1 and E. Type 2, have reciprocal annual cycles of reproduction and nutrient storage. The pyloric caeca (storage organs) reach maximal size over winter then decrease with increasing gametogenic activity in the spring. E. Type 1 broadcasts buoyant eggs in late May to early June. E. Type 2 deposits benthic eggs in late April. Lecithotrophic larval development is similar in both morphs except that larvae of E. Type 1 undergo a brief planktonic phase. E. Type 1 reaches sexual maturity at a larger size, expends a lower reproductive effort but has a greater absolute reproductive output than E. Type 2. E. Type 2 produces fewer, larger eggs and has a greater parental investment per egg. In both morphs, females have much higher gonadal lipid levels than males and expend a higher reproductive effort in terms organic matter and energy. Reproductive effort, reproductive output, nutrient storage in the pyloric caeca and body size varied between populations and between years in each morph. Egg size and parental investment per egg were constant. Within-morph variability is attributed to differences in nutritional state. Differences in reproductive strategy support the hypothesis that the morphs are separate species.     

Influence of hydrodynamic conditions on the production and fate of Posidonia oceanica in a semi-enclosed shallow basin (Stagnone di Marsala,Western Sicily)

An integrated approach using hydrodynamic and transport numerical models, lepidochronology and stable isotope analysis was used to investigate how local hydrodynamic conditions influence the primary production and fate of the seagrass Posidonia oceanica in a Mediterranean semi-enclosed marine system (Stagnone di Marsala). The water mass exchange aptitude of different sectors of the basin was analysed, and data collected were used to select two sectors (colonized by Posidonia oceanica showing the lowest and highest water exchange values) for biological analyses. According to the mean dispersal coefficient differences simulated by the hydrodynamic model, growth rate and primary production of P. oceanica differed between sectors, with average values lower in the central sector where water exchange is lower than in the southern sector. Although P. oceanica coverage and primary production were higher in the southern sector, carbon and nitrogen stable isotope analysis suggests that the transfer of seagrass organic matter to higher trophic levels of the food web was higher in the central sector. The possibility of a link between hydrodynamism, production and fate of organic matter is proposed to explain the observed patterns.