Chloramphenicol influence on antioxidant enzymes with preliminary approach on microsomal CYP1A immunopositive-protein in Chamelea gallina

Chloramphenicol (CA) is a largely used antibiotic and it is an inhibitor of protein synthesis that also induces ROS production. In this work there were investigated activities and expressions in the Adriatic bivalve Chamelea gallina of some antioxidant and detoxification proteins like superoxide dismutase (Mn-SOD, Cu/Zn-SOD), catalase (CAT) and Cytochrome P450 (CYP1A). Clams exposed to 5mgl(-1) of chloramphenicol were sampled 2, 4 and 8 days after treatment (CA2, CA4 and CA8). SODs, CAT, and CYP1A activity and/or expression were detected in pooled digestive glands by Western blotting and by spectrophotometrical analysis. Enzymes activities increase during the entire antibiotic exposure. With respect to the control Cu/Zn-SOD expression increases, while Mn-SOD expression decreases significantly after 4 days. Two CYP1A immunopositive-proteins (57.7 and 59.8kDa) were detected. The lower band significantly decreases in CA8, the upper one also in CA4 condition. High levels of Mn-SOD, CAT activity and Cu/Zn-SOD expression, indicate intense ROS production while Mn-SOD expression inhibition might be ascribable to mitochondrial alterations due to CA and indirectly to ROS. CYP1A1 action determines H(2)O(2) production that would contribute to a CYP1A1 gene promoter down regulation, a response to oxidative stress with the antioxidant enzymes activation as a final result. This study highlights the close association, in C. gallina, in presence of chloramphenicol, between SOD/CAT and CYP system, and it appear particularly interesting to the lack of similar researches on mollusc species.     

Mechanical Property Characterization of Plasticized Sugar Beet Pulp and Poly(Lactic Acid) Green Composites Using Acoustic Emission and Confocal Microscopy

Sorbitol and glycerol were used to plasticize sugar beet pulp-poly(lactic acid) green composites. The plasticizer was incorporated into sugar beet pulp (SBP) at 0%, 10%, 20%, 30% and 40% w/w at low temperature and shear and then compounded with poly(lactic acid) (PLA) using twin-screw extrusion and injection molding. The SBP:PLA ratio was maintained at 30:70. As expected, tensile strength decreased by 25% and the elongation increased. Acoustic emission (AE) showed correlated debonding and fracture mechanisms for up to 20% w/w plasticizer and uncorrelated debonding and fracture for 30–40% sorbitol and 30% glycerol content in SBP–PLA composites. All samples had a well dispersed SBP phase with some aggregation in the PLA matrix. However, at 40% glycerol plasticized SBP–PLA composites exhibited unique AE behavior and confocal microscopy revealed the plasticized SBP and PLA formed a co-continuous two phase system.

Identifying the largest individual sample value from a two-way composite sampling design

Composite sampling techniques for identifying the largest individual sample value seem to be cost effective when the composite samples are internally homogeneous. However, since it is not always possible to form homogeneous composite samples, these methods can lead to higher costs than expected. In this paper we propose a two-way composite sampling design as a way to improve on the cost effectiveness of the methods available to identify the largest individual sample value.     

Unprecedented bleaching-induced mortality in Porites spp. at Rangiroa Atoll, French Polynesia

In April-May 1998, mass coral bleaching was observed in the lagoon of Rangiroa Atoll, Tuamotu Archipelago, French Polynesia. Six months later, the extent of bleaching-induced coral mortality was assessed at three sites. Corals in the fast-growing genus Pocillopora had experienced >99% mortality. Many large colonies of the slow-growing genus Porites (mean horizontal cross-sectional area 5.8 m²) had also died - a phenomenon not previously observed in French Polynesia and virtually unprecedented world-wide. At one site, 25% of colonies, or 44% of the pre-bleaching cover of living Porites, experienced whole-colony mortality. At the two other sites, recently dead Porites accounted for 41% and 82% of the pre-bleaching live cover. Mortality in Porites was negatively correlated with depth between 1.5 and 5 m. Using a 50-year dataset of mean monthly sea surface temperature (SST), derived from ship- and satellite-borne instruments, we show that bleaching occurred during a period of exceptionally high summer SST. 1998 was the first year in which mean monthly SSTs exceeded the 1961-1990 upper 95% confidence limit (29.4°C) for a period of three consecutive months. We suggest that the sustained 3-month anomaly in local summer SST was a major cause of coral mortality, but do not discount the synergistic effect of solar radiation. Recovery of the size-frequency distribution of Porites colonies to pre-bleaching levels may take at least 100 years.     

Elevated levels of nitrogen and phosphorus reduce fertilisation success of gametes from scleractinian reef corals

Spawned gametes were collected from colonies of Acropora longicyathus at One Tree Island and Goniastrea aspera at Magnetic Island, Great Barrier Reef, Australia, for use in fertilisation trials. Mean fertilisation rates were significantly reduced compared with controls (P<0.003), when gametes from the branching coral A. longicyathus were exposed to elevated ammonium concentrations at 1 µM and above in one cross (60-64% reduction), and at 100 µM in another cross (16% reduction). Mean fertilisation success of A. longicyathus gametes was also significantly reduced compared with controls in both crosses (P=0.000) at concentrations of 1 µM phosphate and above (35-75% reduction), and at 1 µM ammonium plus 1 µM phosphate and all higher concentrations (68-74% reduction). Similarly, the mean percentage of regular embryos that were developing normally was significantly reduced in most nutrient treatments compared with controls (P=0.000). Fertilisation trials using gametes from the brain coral G. aspera resulted in a significantly lower percentage of regular embryos (P=0.001) and a significantly higher percentage of deformed embryos (P=0.001) developing after exposure to elevated nutrient treatments compared with controls. Mean fertilisation rates for this species were only significantly reduced (P=0.034) in the 50 µM ammonium plus phosphate treatment in one cross (8% reduction), compared with the control. Therefore, ammonium and phosphate enrichment significantly impairs fertilisation success and embryo development in scleractinian reef corals.     

The Bias of Complementarity Hotspots toward Marginal Populations

Abstract: It has been suggested that using complementarity to identify networks of important areas for conserving biodiversity may preferentially select areas within the margins of species ranges. We tested this idea by examining the location of complementarity hotspots in relation to two measures of range core-periphery. The first measures patterns of aggregation among records within each species' range to identify areas within the core (i.e., areas with aggregated distributions) and periphery (i.e., areas with scattered distributions) of the range. The second measures spatial turnover among species to identify areas with a high density of range edges. For three selected groups of terrestrial vertebrates in Europe—mammals, birds, and herptiles—areas chosen based on complementarity were located within the margins of species' ranges more often than expected by chance. This pattern was consistent for the two measures of core-periphery we used. The bias of complementarity hotspots toward marginal populations is especially important for species with restricted range sizes. If extinctions are determined mainly by demographic factors, then selecting areas at the peripheries of species' ranges might be a poor option. But if extinctions are determined mainly by extrinsic factors, then peripheral populations might be important to ensure the long-term persistence of species.     

Chromosomal and nuclear characteristics of deep-sea hydrothermal-vent organisms: correlates of increased growth rate

A range of tissue and cell types from adult and juvenile stages of vent- and non-vent-dwelling deep-sea and shallow-water organisms were compared for signs of cell division, in preparation for a cytogenetic study of the different groups. Virtually all the Mid-Atlantic Ridge (MAR) vent species (bresiliid shrimp, bathymodiolid mussel, branchipolynoid polychaete, and a range of small gastropods) showed an abundance of metaphase chromosome spreads, indicating a generally high intrinsic rate of cell division, irrespective of their phylogenetic group. In comparison, similar tissues and life-history stages (i.e. gonad, developing eggs and gill) from two deep-sea bathypelagic crustaceans, Hymenodora and Gnathophausia, failed to yield any convincing evidence of cell division. This indicated that rates of growth and reproduction in the rest of the deep sea are extremely low. While much has been written about the contrasting growth rates of vent versus non-vent organisms, this is the first direct evidence of differences in cell-division rate between vent and non-vent species, which undoubtedly reflects contrasting conditions relating to food availability and temporal stability in these two deep-sea environments. The cells of shallow-water marine invertebrates (Mytilus, Nucella, Littorina, Pomatoceros and Crangon), based on the results of previous cytogenetic investigations, occupy an intermediate position with respect to their cell-division kinetics. When chromosome numbers, DNA content and nuclear diameters were compared between vent and non-vent species, this showed that no change had occurred in these characters since the time of vent colonisation. Representative chromosome spreads from hydrothermal-vent organisms are shown here for the first time.     

Hepatopancreatic endosymbionts in coastal isopods (Crustacea: Isopoda), and their contribution to digestion

Three isopod species (Crustacea: Isopoda), commonly found in the intertidal and supratidal zones of the North American Pacific coast, were studied with respect to symbiotic microbiota in their midgut glands (hepatopancreas). Ligia pallasii (Oniscidea: Ligiidae) contained high numbers of microbial symbionts in its hepatopancreatic caeca. Numbers of endosymbionts were strongly reduced by ingestion of antibiotics. By contrast, the hepatopancreas of Idotea wosnesenskii (Valvifera: Idoteidae) and Gnorimosphaeroma oregonense (Sphaeromatidea: Sphaeromatidae) did not contain any microbiota. Results of feeding experiments suggest that microbial endosymbionts contribute to digestive processes in L. pallasii, the most terrestrial of the three isopods that we studied. The acquisition of digestion-enhancing endosymbionts may have been an important evolutionary step allowing isopods to colonize terrestrial habitats where relatively indigestible leaf litter is the primary food source. By contrast, the ability to digest phenolic compounds was most developed in one of the more marine species, suggesting that this trait may have evolved independently in isopod species that consume a phenolic-rich diet, whether in marine habitats or on land. Received: 28 August 2000 / Accepted: 8 December 2000     

Molecular response of the sponge Suberites domuncula to bacterial infection

The aim of this study was the documentation of the molecular immune response of Suberites domuncula upon bacterial infection. Additionally, the bacteria that are naturally present in the sponge after prolonged aquarium maintenance were characterized. After 6 months of maintenance of S. domuncula in seawater aquaria, only one bacterial 16S rDNA sequence could be recovered, which belongs to the genus Pseudomonas. Concomitantly, morphologically uniform bacteria were found encapsulated in bacteriocytes. These findings indicate that certain bacteria, possibly of the genus Pseudomonas, are able to persist for long periods in host bacteriocytes. Subsequent to performing a previously established infection assay with S. domuncula, a potentially pathogenic Vibrio sp. was isolated from the tissues. Furthermore, the host tissue disintegrated and asexual propagation bodies (gemmules) were formed. In order to gain insights into the molecular events occurring after bacterial infection, the stress-response kinases, p38 protein kinase and JNK protein kinase, were analyzed. It is demonstrated that these two kinases are activated (phosphorylated) upon incubation of the tissue with the bacterial endotoxin lipopolysaccharide (LPS). Moreover, LPS strongly inhibits protein synthesis. It is concluded that there are many functionally different interactions between S. domuncula and bacteria and that the animal possesses mechanisms to differentiate between bacteria and to respond accordingly.