Increase in Alphaproteobacteria in association with a polychaete,Capitella sp. I,in the organically enriched sediment

We conducted bioremediation experiments on the organically enriched sediment on the sea floor just below a fish farm, introducing artificially mass-cultured colonies of deposit-feeding polychaete, Capitella sp. I. To clarify the association between the Capitella and bacteria on the efficient decomposition of the organic matter in the sediment in the experiments, we tried to identify the bacteria that increased in the microbial community in the sediment with dense patches of the Capitella. The relationship between TOC and quinone content of the sediment as an indicator of the bacterial abundance was not clear, while a significant positive correlation was found between Capitella biomass and quinone content of the sediment. In particular, ubiquinone-10, which is present in members of the class Alphaproteobacteria, increased in the sediment with dense patches of the Capitella. We performed denaturing gradient gel electrophoresis (DGGE) analyses to identify the alphaproteobacterial species in the sediment with dense patches of the worm, using two DGGE fragments obtained from the sediment samples and one fragment from the worm body. The sequences of these DGGE fragments were closely related to the specific members of the Roseobacter clade. In the associated system with the Capitella and the bacteria in the organically enriched sediment, the decomposition of the organic matter may proceed rapidly. It is very likely that the Capitella works as a promoter of bacteria in the organically enriched sediment, and feeds the increased bacteria as one of the main foods, while the bacteria decompose the organic matter in the sediment with the assistance of the Capitella.     

Effects of sediment-bound Cd, Pb, and Ni on the growth, feeding, and survival of Capitella sp. I

Anthropogenic metal pollutants bioaccumulated in benthic animals by means of feeding and osmotic diffusion. These metals may affect the physiology of the benthos. In this study, we exposed Capitella sp. I to three metals (Cd, Pb, and Ni), each in eight different concentrations, to determine the effects of metals on the animals. Growth rate, ingestion rate, and percent survival were estimated in three separated experiments. The growth and feeding of the worms were sensitive to even the lowest concentrations of each metal added to the sediments. The lowest observable adverse effect levels for Cd, Ni, and Pb were 0.03, 1.59, and 0.41 μmol g^− 1 sediment, respectively. Growth rates in the elevated metal contaminant treatments decreased drastically at slightly contaminated levels, lessened detrimental effects at moderately contaminated levels, and showed incompensable intoxication at heavily contaminated levels. The trends in ingestion rates were similar to those of growth rates. No significant difference in survivorship was found among the different contaminant levels for any of the three heavy metals. Capitella sp. I was most sensitive to Cd, followed by Ni and Pb, which had similar effects. The rapid physiological responses of Capitella sp. I allowed the animals to survive metal exposure. Sediment productivity remained unchanged at different contamination levels of Ni and Pb, but was drastically reduced at 4.75 μmol g^− 1 Cd in the sediment. This further demonstrated Capitella sp. I can adjust their ingestion rates to maintain constant sediment productivities in moderate pollution conditions; however, when threshold concentration was exceeded, homeostasis collapsed.     

Morphological features and viability of Scrippsiella trochoidea cysts isolated from fecal pellets of the polychaete Capitella sp.

To investigate the impact of grazing by the polychaete Capitella sp. on the two cyst morphotypes of Scrippsiella trochoidea, the typical morphotype with short calcareous spines (spiny-type cyst) and artificially induced the transparent type without calcareous spines (naked-type cyst), we examined the morphological features and germination capability of the two cyst morphotypes isolated from fecal pellets of the polychaete Capitella sp. produced in a restricted habitat. The morphological destruction was observed in both spiny- and naked type cysts after passage through the gut of Capitella sp., and this seemed to occur rapidly for naked-type cysts. In addition, the germination of both spiny- and naked-type cysts isolated from fecal pellets on day 2 of harvesting was significantly reduced and subsequently completely abolished, in contrast to previous findings from ingestion studies. Our results indicate that continual grazing by Capitella sp. within a restricted habitat can compromise the survival of S. trochoidea cysts.     

The use of growth and ingestion rates of Capitella sp. I as the bioassay approaches to determine the sediment quality of coastal wetlands of Taiwan

Food is a limiting factor for the deposit feeders. The availability of sediment nutrients thus has a tight relationship with the growth, survival and development of the animal. There are two purposes of this study: (1) to determine if the ingestion and growth rates can be used as a bioassay approach to assess the sediment nutrients; and (2) use the combination of bioassay approaches and chemical analyses to determine which chemical parameter is the better predicator of the sediment nutrients to the animals. In the preliminary study, the optimal growth length and average ingestion rate of Capitella sp. I were obtained from the laboratory. The standardized relationships of the growth and ingestion rates in response to different nutrients were prepared. Then, the sediments collected from different coastal wetlands in Western Taiwan were used in the feeding, growth experiments and chemical analyses. The comparisons were made between the field and laboratory experiments to determine the sediment nutrients in the wetland of Taiwan. In the growth rate standardized relationship, Capitella sp. I increased its growth rate with the total organic nitrogen (TON) concentration between 0 to 2.8 mgN·g sediment⁻¹, total organic carbon (TOC) concentration between 0 and 22.4 mgC·g sediment⁻¹, and enzymatically hydrolyzable amino acid (EHAA) concentration between 0 and 4.48 mg protein·g sediment⁻¹. After the nutrient concentrations exceed these values, the growth rates decreased gradually. In the ingestion rate standardized relationship, the animal increased its ingestion rate with the total organic nitrogen (TON) concentration between 0 and 2 mgN·g sediment⁻¹, total organic carbon (TOC) concentration between 0 and 14.1 mgC·g sediment⁻¹, and EHAA concentration between 0 and 3.2 mg protein·g sediment⁻¹. After the nutrient concentrations exceed these values, the ingestion rates also decreased. To determine which nutrient parameter is the best predictor for the sediment nutrient in the field, we first analyzed whether the data obtained from the laboratory fell within 99% confidence interval of the regression obtained from the field data. Then, to determine which parameter had the shortest perpendicular distance between the field and the laboratory regression curves. Both the growth and ingestion rates comparisons showed that the EHAA is the best candidate of the sediment nutrient of deposit feeders in the field. The results of this study proved tentatively that the growth and ingestion rates of Capitella sp. I can be used as the bioassay approaches to estimate the sediment nutrients. The combination of the bioassay approaches and the relevant chemical analyses allows us to determine the bioavailability fraction of sediment to the deposit feeders.     

Chemical induction of settlement and metamorphosis of Capitella capitata Sp. I (Polychaeta) larvae by juvenile hormone-active compounds

Summary

The influence of juvenile hormone (JH)-active chemicals on the settlement and metamorphosis of metatrochophore larvae of the polychaete annelid Capitella sp. I of the Capitella complex has been investigated. These studies demonstrate that JH-active chemicals are able to induce settlement and metamorphosis of Capitella larvae, and that these effects may possibly be mediated by protein kinase C induction. Evidence for the presence of JH-active compounds in marine sediments is also presented, suggesting that these chemicals may serve a natural role as chemical cues for settlement and metamorphosis for Capitella larvae in the marine environment.     

The role of sediment organic matter composition in larval habitat selection by the polychaete Capitella sp. I

The role of sediment organic matter composition in larval habitat selection by the polychaete Capitella sp. I was investigated using a still water multiple-choice experiment. Specifically, the following question was addressed: what are the roles of sediment labile organic compounds in larval habitat selection? A replicated Latin-square design was used to test larval preferences among sediments collected from five sites (TLC—Tung Lung Chau; VHE—Victoria Harbor east; VH—Victoria Harbor; VHW—Victoria Harbor west; PC—Peng Chau) in the vicinity of Victoria Harbor, Hong Kong. Although mean grain size was not different among sites, water content and the total organic matter were higher in VH Mud but were similar among other sites. Carbohydrate, lipid, and biopolymeric fractions followed the same trend. According to protein content, sites were ranked as: VH > VHW > TLC = VHE > PC. PC Mud had the highest carbohydrate/protein ratio, due to low concentration of protein. According to larval preference, sites were ranked as: TLC = VH = VHW = VHE > PC. Our study shows a precise relationship between sediment biochemical properties and larval habitat selection. Larvae tend to choose sediments with low carbohydrate/protein ratio. Results suggest that types of sediment organic matter may play more important role in the larval habitat selection process of Capitella sp. I than concentration of organic matter alone.     

Size-specific effects of increased sediment loads on gastropod communities in Lake Tanganyika, Africa

The remarkable biodiversity of the littoral zone of Lake Tanganyika appears to be at risk through increasing sediment input caused by anthropogenic pressures. An in-situ field experiment was done to investigate the effects of increased sediment loads on the size-structure of gastropod communities on a rocky shore site in the lake. Gastropods were removed prior to the addition of sediment, and subsequent further removals were done after seven days and six months. Relative to controls, mean size of individuals of both Lavigeria grandis (Smith) and Lavigeria sp. Q decreased following the addition of sediment, while that of Lavigeria sp. P increased. A subsequent laboratory experiment found that survivorship of larger individuals of Lavigeria grandis was less than smaller individuals in the sediment-impacted treatment. It is hypothesised that decreased algal availability on sediment-impacted rock surface increased starvation of larger snails. Alterations to the size-structure of Lavigeria sp. P populations appear related to intraspecific differences in competitive ability. These results highlight the complex and unpredictable effects of increases in sediment loads on aquatic ecosystems and have implications for the conservation of littoral communities in sediment-impacted areas.     

Temporal and spatial patterns of annelid populations in intertidal sediments of the Quequén Grande estuary (Argentina)

Temporal and spatial patterns of polychaete and oligochaete populations inhabiting intertidal sediments of the Quequén Grande estuary (Buenos Aires Province, Argentina) were analysed monthly during a yearly period. Local sediments are poorly selected, very fine sands, with a high percentage of mud. Two sites, located at both banks of the estuary were studied: (1) Site N, with a comparatively steep slope and percentage of mud increasing with sediment depth; (2) Site Q, with a gentle slope and percentage of mud increasing towards the sediment surface. The assemblage of annelids was characterised by a very low specific richness, being composed just by 4 species: the polychaetes Laeonereis acuta, Boccardiella ligerica and Capitella sp., and the tubificid oligochaete Ilyodrilus cf. frantzi. Density of the nereid L. acuta reached a peak in late summer and a minimum in spring. The population of Capitella sp. also showed maximum densities in summer, but collapsed during early fall, disappearing completely from the study area. Densities of I. cf. frantzi were highly variable in time and space. Three of the 4 species showed maximum densities at site Q, where mud content was highest at the sediment surface. At this sampling site, most of the L. acuta population occurred within the upper 8 cm of the substrate. On the contrary, at site N, where mud and organic matter content increased with depth, this species was more abundant at subsurface layers (8–16 cm) than at the sediment surface.     

Cytogenetic and genotoxic effects of the insecticides, imidacloprid and methamidophos

We examined the cytogenetic and genotoxic effects of the neonicotinoid insecticide imidacloprid and the organophosphate insecticide methamidophos, when administered alone or in combination. These insecticides were tested with the bone marrow chromosome aberration assay and micronucleus test in rats and by the bacterial mutation assay (Salmonella/microsome mutagenicity assay). Wistar albino rats were orally fed daily with laboratory chow treated with various concentrations of insecticides, 50 and 100 mg/kg imidacloprid, 2.5 and 5 mg/kg methamidophos, and 2.5 and 5 mg/kg imidacloprid plus methamidophos, respectively, for 90 days. Numerical and structural chromosomal aberrations were evaluated. Significant differences were detected between all the insecticide-administered groups versus the control group and between the two concentrations of the pesticide-treated groups. Both concentrations of the insecticides induced a dose-related increase in the micronucleus frequency (P < 0.05). Dose-related increases in the number of revertants were observed with the two Salmonella strains (TA98 and TA100). All tested doses of the insecticides demonstrated mutagenic activity in the presence of S9 mix. These results lead us to the conclusion that the synergistic effect of methamidophos and imidacloprid causes an increase in potential damage to non-target organisms.     

Benthic fauna bio-irrigation effects on nutrient regeneration in fish farm sediments

Impacts of organic enrichment and a modified benthic fauna community (caused by fish farming) on benthic mineralization rates and nutrient cycling were studied in sediments at one Danish and one Cypriote fish farm. Sediment organic matter concentration and macrofauna community composition were manipulated in microcosms and changes in total benthic metabolism (oxygen consumption, TCO₂ production), anaerobic metabolism (sulfate reduction rates), nutrient fluxes and sediment parameters were followed for a period of 3 weeks. Mineralization rates were found to be highly correlated with irrigation velocities and largest fauna effects were found in the Danish sediments with the large and active irrigating climax species (Nereis diversicolor and Macoma balthica). Eastern Mediterranean climax species (Glycera rouxii and Naineris laevigata) also stimulated mineralization rates but to a smaller extent due to lower irrigation, whereas the opportunistic species (Capitella in Danish sediment and Hermodice carunculata in Cypriote sediment) showed less effect on mineralization. Ammonium and phosphate release increased with increasing irrigation velocities, but much less in Cyprus indicating higher nutrient retention at the ultra-oligotrophic location compared to eutrophic Danish site. Irrigation velocities, and thus mineralization rates, increased by organic matter loading, indicating larger fauna-induced oxidation in enriched environments. The result implies that a change in fauna structure in fish farm sediment towards smaller opportunistic polychaete species with lower irrigation will result in slower mineralization rates and potentially increase accumulation of organic waste products.     

Short-term response of deposit-feeders to an increase of the nutritive value of the sediment through seasons in an intertidal mudflat (Western Mediterranean, Italy)

In coastal intertidal flats, assemblages of deposit-feeders often vary seasonally and according to the availability of food in sediments. In the mudflat where this study has been done, the deposit-feeder assemblage changed after a sharp increase of fresh detritus, which occurred in summer 1997. The number of the animals increased and abundances of some worms (Streblospio shrubsolii, Desdemona ornata and Capitella cf. capitata) were correlated with the nutritive value of the sediment. In addition, there was evidence that increasing food in summer would increase recruitment of juveniles and adults of S. shrubsolii.In this paper, the concentration of food was manipulated in situ in sediments of an intertidal mudflat in different seasons. Food of high nutritive value was added to small plots of sediments to test hypotheses about effects on the distribution of deposit-feeders. It was predicted that addition of food would change composition and abundance of the assemblage and densities of the deposit-feeders that were correlated to the availability of food. Furthermore, it was tested whether juveniles and adults of S. shrubsolii were affected by the availability of food consistently through seasons.Densities of deposit-feeders increased in plots where the sediment was organically enriched. The composition of the assemblage, however, was not affected. S. shrubsolii, D. ornata and C. cf. capitata responded to the addition of food. However, the nutritive value of the sediment affected the dynamics of these animals variably through seasons. Abundances increased in the experimental treatment E in winter and spring for D. ornata and in winter and autumn for C. cf. capitata. Furthermore, in summer and autumn juveniles and adults of S. shrubsolii were more abundant in plots where food was added.     

The complex effects of the invasive polychaetes Marenzelleria spp. on benthic nutrient dynamics

The effects of the polychaetes Marenzelleria sp. (Polychaeta, Spionidae), nonindigenous, rapidly increasing species in the Baltic Sea, on benthic nutrient fluxes, denitrification and sediment pore water nutrient concentration were studied in laboratory experiments using a flow-through setup with muddy sediment from coastal regions of the Gulf of Finland. In addition, different forms of sediment phosphorus (P), separated by chemical fractionation, were studied in three sediment layers. At a population density corresponding to about half the highest measured in the northern Baltic Sea, Marenzelleria sp. increased the fluxes of P and ammonium to the water column. No effect could be recorded for denitrification. Since the previously dominant species of the area, Monoporeia affinis, can enhance denitrification and lower the amount of dissolved P in the pore water, the replacement of M. affinis with Marenzelleria spp. may lead to increased P flux to the water column and decreased denitrification, further increasing the ammonium flux to the water column. However, sediment reworking by Marenzelleria spp. also oxidizes the surface sediment in the long run, improving its ability to retain P and support nitrification. Therefore, the impact of Marenzelleria spp. on sediment nutrient release may not be as drastic as the initial reactions seen in our experiments suggest.     

Positive correlation of methamidophos resistance between Lipaphis erysimi and Diaeretilla rapae and effects of methamidophos ingested by host insect on the parasitoid

Temporal and spatial correlated variations on the methamidophos resistance and biomolecular rate constant of acetylcholinesterase to insecticides were found between the turnip aphid, Lipaphis erysimi and its endoparasitoid, Diaeretilla rapae , collected from field colonies and an insecticide-free field insectarium in Fujian, China. Compared to the related susceptible insectarium population, L. erysimi and D. rapae displayed 7.4–29.2- and 2.6–9.2-fold resistance ratios, respectively. In addition, two populations of L. erysimi with different methamidophos resistance levels, that is, a field (with 5.8-fold resistance ratio) and an insectarium population, were used to study the effects of methamidophos ingested by the host insect on D. rapae development. The percentage of D. rapae cocoon formation decreased significantly when the parasitized L. erysimi were fed on cauliflower leaves treated with methamidophos at lethal concentration dosages 10 (LC₁₀) or LC₅₀. At LC₅₀ dosages the percentage of D. rapae cocoon formation and adult emergence decreased significantly. When the parasitized L. erysimi were fed on methamidophos at LC₉₀ dosage, no D. rapae cocoons were found. When the field or insectarium L. erysimi were treated with methamidophos at LC₁₀, the susceptibility to methamidophos in the adult D. rapae emerged from the treated host insect was similar to the control. However, the susceptibility to methamidophos in the adult D. rapae became lower than the control when the host insects were treated at LC₅₀ dosages. The data thus suggested that the methamidophos ingested by the host insect L. erysimi could be an important factor in the endoparasitoids' insecticide resistance development. The natural selectivity would favor the parasitoids that had developed an insensitivity to the insecticide(s).     

Insecticide odour interference with food-searching behaviour of Microplitis croceipes (Hymenoptera: Braconidae) in a laboratory arena

Response to odours plays an important role in resource location by natural enemies, particularly by parasitoid wasps. While a considerable research effort has been dedicated to studying the effects of insecticide intoxication on natural enemy search behaviour, it is yet unknown if the odours themselves interfere with distant chemoreception. We investigated this issue using the food-searching behaviour of Microplitis croceipes (Cresson)(Hymenoptera: Braconidae) in laboratory arenas as a model system. Odours of imidacloprid (Genesis®), spinosad (Entrust®), esfenvalerate (Asana®), methamidophos (Monitor®), and vanilla were tested for their ability to interfere with wasp response to the odour of honey. The wasps did not contact the chemicals. Honey odour was generally effective in triggering food-searching behaviour in both honey-fed (and thus conditioned to associate honey odour with food) and unfed, naïve wasps. Mixing honey with imidacloprid and spinosad did not affect wasp responses. The remaining compounds (esfenvalerate, methamidophos, and vanilla) significantly reduced the proportion of positively responding fed wasps, but only methamidophos had such an effect on the unfed wasps. Negative methamidophos effects became completely reversed when wasps were forced to feed on honey in the presence of methamidophos odour. Our results suggest that odours and provision of food may potentially be used to keep beneficial natural enemies away from insecticide-treated areas.     

New marine mammal faunas (Cetacea and Sirenia) and sea level change in the Samlat Formation,Upper Eocene,near Ad-Dakhla in southwestern Morocco

The Samlat Formation is well exposed in coastal sections bordering the Atlantic Ocean south of Ad-Dakhla in southwestern Morocco. Here some 22 m of rhythmically-bedded, chert-rich, marine siltstones and marls are overlain by 1–1.5 m of vertebrate-bearing microconglomeratic sandstone, another 4–8 m of rhythmically-bedded siltstone and marl, and finally a second 3–6 m unit of vertebrate-bearing muddy sandstone. The microconglomeratic and muddy sandstones represent low sea stands in what is otherwise a deeper water sequence. Cetacean skeletons are rare but cetacean vertebrae are common in the lower sandstone (bed B1), where many show the effects of reworking. The cetaceans in bed B1represent a minimum of five species, from smallest to largest: cf. Saghacetus sp., cf. Stromerius sp., Dorudon atrox, cf. Dorudon sp., and Basilosaurus isis. Bed B1 yields rib fragments that may represent sirenians, but sirenians, if present, are rare. The only identifiable cetacean found in the upper sandstone (bed B2) is Basilosaurus sp. Dugongid sirenians identified as cf. Eosiren sp. are the most common mammal in bed B2. We interpret co-occurrence of the typically Early Priabonian species Dorudon atrox and Basilosaurus isis with smaller species more like Middle Priabonian genera Saghacetus osiris and Stromerius nidensis to indicate that bed B1 was deposited during low sea stand Pr-2 between the Early and Middle Priabonian (between the early and middle Late Eocene). Bed B2 is separated from B1 by an interval of deeper water sediment accumulation. Bed B2 could represent a later phase of Pr-2 or a subsequent Priabonian low sea stand (possibly Pr-3).     

Pretrichodermamide C and N-methylpretrichodermamide B,two new cytotoxic epidithiodiketopiperazines from hyper saline lake derived Penicillium sp.

Two new epidithiodiketopiperazines, pretrichodermamide C (1) and N-methylpretrichodermamide B (2) were isolated from the fungus Penicillium sp., derived from the sediment of a hyper saline lake located at Wadi El-Natrun in Egypt. The structures of 1 and 2 were unambiguously determined on the basis of one- and two-dimensional NMR spectroscopy and by high-resolution mass spectrometry, as well as by comparison with the literature. Compound 2 showed pronounced cytotoxicity against the murine lymphoma L5178Y cell line with an IC₅₀ value of 2 μM.     

Characterization of <Emphasis Type="Italic">twist</Emphasis> and <Emphasis Type="Italic">snail</Emphasis> gene expression during mesoderm and nervous system development in the polychaete annelid <Emphasis Type="Italic">Capitella</Emphasis> sp. I

To investigate the evolutionary history of mesoderm in the bilaterian lineage, we are studying mesoderm development in the polychaete annelid, Capitella sp. I, a representative lophotrochozoan. In this study, we focus on the Twist and Snail families as candidate mesodermal patterning genes and report the isolation and in situ expression patterns of two twist homologs (CapI-twt1 and CapI-twt2) and two snail homologs (CapI-sna1 and CapI-sna2) in Capitella sp. I. CapI-twt1 is expressed in a subset of mesoderm derivatives during larval development, while CapI-twt2 shows more general mesoderm expression at the same stages. Neither twist gene is detected before the completion of gastrulation. The two snail genes have very distinct expression patterns. At cleavage and early gastrula stages, CapI-sna1 is broadly expressed in precursors of all three germ layers and becomes restricted to cells around the closing blastopore during late gastrulation; CapI-sna2 expression is not detected at these stages. After gastrulation, both snail genes are expressed in the developing central nervous system (CNS) at stages when neural precursor cells are internalized, and CapI-sna1 is also expressed laterally within the segmental mesoderm. Based on the expression patterns in this study, we suggest a putative function for Capitella sp. I twist genes in mesoderm differentiation and for snail genes in regulating CNS development and general cell migration during gastrulation. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Spatial and Temporal Potato Intensification Drives Insecticide Resistance in the Specialist Herbivore,Leptinotarsa decemlineata

Landscape-scale intensification of individual crops and pesticide use that is associated with this intensification is an emerging, environmental problem that is expected to have unequal effects on pests with different lifecycles, host ranges, and dispersal abilities. We investigate if intensification of a single crop in an agroecosystem has a direct effect on insecticide resistance in a specialist insect herbivore. Using a major potato pest, Leptinotarsa decemlineata, we measured imidacloprid (neonicotinoid) resistance in populations across a spatiotemporal crop production gradient where potato production has increased in Michigan and Wisconsin, USA. We found that concurrent estimates of area and temporal frequency of potato production better described patterns of imidacloprid resistance among L. decemlineata populations than general measures of agricultural production (% cropland, landscape diversity). This study defines the effects individual crop rotation patterns can have on specialist herbivore insecticide resistance in an agroecosystem context, and how impacts of intensive production can be estimated with general estimates of insecticide use. Our results provide empirical evidence that variation in the intensity of neonicotinoid-treated potato in an agricultural landscape can have unequal impacts on L. decemlineata insecticide insensitivity, a process that can lead to resistance and locally intensive insecticide use. Our study provides a novel approach applicable in other agricultural systems to estimate impacts of crop rotation, increased pesticide dependence, insecticide resistance, and external costs of pest management practices on ecosystem health.