Chronic toxicity of ibuprofen to Daphnia magna: Effects on life history traits and population dynamics

The non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) is a widely used pharmaceutical that can be found in several freshwater ecosystems. Acute toxicity studies with Daphnia magna suggest that the 48 h EC50 (immobilisation) is 10-100 mg IB l(-1). However, there are currently no chronic IB toxicity dataon arthropod populations, and the aquatic life impacts of such analgesic drugs are still undefined. We performed a 14-day exposure of D. magna to IB as a model compound (concentration range: 0, 20, 40 and 80 mg IB l(-1)) measuring chronic effects on life history traits and population performance. Population growth rate was significantly reduced at all IB concentrations, although survival was only affected at 80 mg IB l(-1). Reproduction, however, was affected at lower concentrations of IB (14-day EC50 of 13.4 mg IB l(-1)), and was completely inhibited at the highest test concentration. The results from this study indicate that the long-term crustacean population consequences of a chronic IB exposure at environmentally realistic concentrations (ng l(-1) to mu g l(-1)) would most likely be of minor importance. We discuss our results in relation to recent genomic studies, which suggest that the potential mechanism of toxicity in Daphnia is similar to the mode of action in mammals, where IB inhibits eicosanoid biosynthesis. (C) 2007 Elsevier Ireland Ltd. All rights reserved.

Temporal and spatial variability of mobile fauna on a submarine cliff and boulder scree complex: a community in flux

The mobile component of a community inhabiting a submarine boulder scree/cliff was investigated at Lough Hyne, Ireland at dawn, midday, dusk and night over a 1-week period. Line transects (50 m) were placed in the infralittoral (6 m) and circumlittoral (18 m) zones and also the interface between these two zones (12 m). The dominant mobile fauna of this cliff consisted of echinoderms (6 species), crustaceans (10 species) and fish (23 species). A different component community was identified at each time/depth interval using Multi-Dimensional Scaling (MDS) even though both species diversity (Shannon-Wiener indices) and richness (number of species) remained constant. These changes in community composition provided indirect evidence for migration by these mobile organisms. However, little evidence was found for migration between different zones with the exception of the several wrasse species. These species were observed to spend the daytime foraging in the deeper zone, but returned to the upper zone at night presumably for protection from predators. For the majority of species, migration was considered to occur to cryptic habitats such as holes and crevices. The number of organisms declined during the night, although crustacean numbers peaked, while fish and echinoderms were most abundant during day, possibly due to predator-prey interactions. This submarine community is in a state of flux, whereby, community characteristics, including trophic and energetic relationships, varied over small temporal (daily) and spatial (m) scales.

ATP pulse and calcium homeostasis in cells from hepatopancreas of Dilocarcinus pagei, a freshwater crab

Calcium (Ca) is critical for crustaceans due to their molting cycle and its presence in the carapace as calcium carbonate, apart from the usual functions of Ca, such as cell signalling. Ca transport in Dilocarcinus pagei, a freshwater crab, was studied in isolated cells from hepatopancreas to further characterize Ca transport mechanisms in these crabs. Cells were isolated and loaded with Fluo-3, a calcium fluorescent dye. Three different cell treatments were performed: Group 1 cells were Ca free during cell dissociation, and calcium was present (at 1mM) for fluorescence cell loading and transport experiments (FC); Group 2 cells were calcium free during cell dissociation and for transport experiments, but not during cell loading (LC); and Group 3 cells were Ca free during cell dissociation, cell loading and transport experiments (WC). Intracellular Ca was recorded through time after ATP was added to the cells and ATP caused an increase in Ca efflux within 30s in all cells. WC cells showed the smallest Ca efflux compared to the other cells, probably because it was intracellularly Ca ""depleted"". Vanadate and amiloride decreased the Ca efflux when ATP was added to the cells, while verapamil did not cause any effect in Ca efflux, confirming the presence of a Ca(2+)-ATPase sensitive to vanadate in hepatopancreas of D. pagei. In a different set of experiments, cells were also exposed to a Ca pulse of 1 and 10mM during 180s. 10mM Ca increased intracellular Ca compared to 1mM, and the increase was not recovered during the experimental time. Additionally, Ca influx was reduced by verapamil and amiloride, but not completely. The results suggest that Ca influx probably occurs through an undefined exchanger, apart from Ca channels (verapamil sensitive) and electrogenic 1Na(+)(1H(+))/1 Ca(2+) exchanger (amiloride-sensitive). Similarities between freshwater and seawater crabs, lobsters and crayfish in relation to plasma membrane Ca transporters, although the environment where they live is quite diverse, suggest that universal mechanisms for Ca homeostasis are widespread among crustaceans. (C) 2010 Elsevier Inc. All rights reserved.

Differential effects of alpha-helical and beta-hairpin antimicrobial peptides against Acanthamoeba castellanii

In this work we evaluated the ability of different types of antimicrobial peptides to promote permeabilization and growth inhibition of Acanthamoeba castellanii trophozoites, which cause eye keratitis. We used cationic alpha-helical peptides P5 and a beta-hairpin amphipathic molecule (gomesin), of the spider Acanthoscurria gomesiana haemocytes. A. castellanii permeabilization was obtained after 1 h incubation with micromolar concentrations of both types of peptides. While permeabilization induced by gomesin increased with longer incubations, P5 permeabilization did not increase with time and occurred at doses that are more toxic for SIRC cells, P5, however, at doses below the critical dose used to kill rabbit corneal cells was quite effective in promoting growth inhibition. Similarly, P5 was more effective when serine protease inhibitor was added simultaneously to the permeabilization assay. High performance chromatography followed by mass spectrometry analysis confirmed that, in contrast to gomesin, P5 is hydrolysed by A. castellanii culture supernatants. We conclude that the use of antimicrobial peptides to treat A. castellanii infections requires the search of more specific peptides that are resistant to proteolysis.

Single Synapse Information Coding in Intraburst Spike Patterns of Central Pattern Generator Motor Neurons

Burst firing is ubiquitous in nervous systems and has been intensively studied in central pattern generators (CPGs). Previous works have described subtle intraburst spike patterns (IBSPs) that, despite being traditionally neglected for their lack of relation to CPG motor function, were shown to be cell-type specific and sensitive to CPG connectivity. Here we address this matter by investigating how a bursting motor neuron expresses information about other neurons in the network. We performed experiments on the crustacean stomatogastric pyloric CPG, both in control conditions and interacting in real-time with computer model neurons. The sensitivity of postsynaptic to presynaptic IBSPs was inferred by computing their average mutual information along each neuron burst. We found that details of input patterns are nonlinearly and inhomogeneously coded through a single synapse into the fine IBSPs structure of the postsynaptic neuron following burst. In this way, motor neurons are able to use different time scales to convey two types of information simultaneously: muscle contraction (related to bursting rhythm) and the behavior of other CPG neurons (at a much shorter timescale by using IBSPs as information carriers). Moreover, the analysis revealed that the coding mechanism described takes part in a previously unsuspected information pathway from a CPG motor neuron to a nerve that projects to sensory brain areas, thus providing evidence of the general physiological role of information coding through IBSPs in the regulation of neuronal firing patterns in remote circuits by the CNS.

Diphtheria toxoid conformation in the context of its nanoencapsulation within liposomal particles sandwiched by chitosan

Chitosan (alpha alpha-(1-4)-amino-2-deoxy-beta beta-D-glucan) is a deacetylated form of chitin, a polysaccharide from crustacean shells. Its unique characteristics, such as positive charge, biodegradability, biocompatibility, nontoxicity, and rigid structure, make this macromolecule ideal for an oral vaccine delivery system. We prepared reverse-phase evaporation vesicles (REVs) sandwiched by chitosan (Chi) and polyvinylic alcohol (PVA). However, in this method, there are still some problems to be circumvented related to protein stabilization. During the inverted micelle phase of protein nanoencapsulation, hydrophobic interfaces are expanded, leading to interfacial adsorption, followed by protein unfolding and aggregation. Here, spectroscopic and immunological techniques were used to ascertain the effects of the Hoffmeister series ions on diphtheria toxoid (Dtxd) stability during the inverted micelle phase. A correlation was established between the salts used in aqueous solutions and the changes in Dtxd solubility and conformation. Dtxd alpha alpha-helical content was quite stable, which led us to conclude that encapsulation occurred without protein aggregation or without exposition of hydrophobic residues. Dtxd aggregation was 98% avoided by the kosmotropic, PO

Projeto raia de fogo: aspectos populacionais da raia de fogo (Dasyatis marianae Gomes, Rosa & Gadig, 2000) e pesca de elasmobrânquios no complexo recifal do Parracho de Maracajaú

The species Dasyatis marianae inhabits coastal areas associated with coral reefs, considered endemic to the northeast of Brazil, occurring from the State of Maranhão to the south of Bahia. Specimens of this species are commonly sighted by divers and fishermen in the area of Maracajaú reefs, a complex reef that is part of the Environmental Protection Area of Coral Reefs (EPACR), which was developed in this study about the ecology and biology of the D. marianae, in order to characterize aspects of population structure in the area of the reef complex of Parracho de Maracajaú. We analyzed 120 specimens caught by artisanal fishing site of the size, weight, sex, stage of maturity and stomach contents. Most subjects were adult males (1.7:1) and was more abundant for rays with lengths between 25 and 29cm of LD, where females reach larger sizes, a feature common to other rays. The largest specimens were captured in the area of seagrass, which is preferred for the species. The distribution of species in the area showed an ontogenetic and sexual segregation, where the youthful occur near the beach, which is a likely area for nursery and growth of the adult females prevail in the seagrass, which apparently has a high prey availability, and Adult males are more distant, a higher proportion occurring in outlying areas, suggesting a habit more exploratory than the females. The evaluation of the reproductive system indicated 3 reproductive cycles per year, one young per pregnancy, and showed that the mature males were smaller than females. The cubs of D. marianae size at birth 12 to 15cm LD. As for diet, the species was characterized as carnivorous crustacean specialist. The performance of visual censuses in different localities allowed to evaluate the density of D. marianae in different environments of the complex. The species occurs in greater numbers in seagrass, environment very important for the conservation of the species. 100 individuals of D. marianae marked in reef complex area enrolled in a recapture rate of 3%. Some behavioral aspects were evaluated, as diurnal pattern of activity, interaction with cleaning and fish Pomacanthus paru followers as Lutjanus analis and Carangoides bartholomaei. Overall, much of the information obtained should be used for management of the species

Atividade β-D-N-Acetilglucosaminidásica de enzimas imobilizadas extraídas da Artemia franciscana e possíveis aplicações biotecnológicas

A β-D-N-acetilglucosaminidase extracted and partially isolated from crustacean Artemia franciscana by ammonium sulfate precipitation and filtration gel chromatography Bio Gel A 1.5m. the enzyme was immobilized on ferromagnetic Dacron yielding a insoluble active derivative with 5.0 units/mg protein and 10.35% of the soluble enzyme activity. β-D-N-acetilglucosaminidase-ferromagnetic Dacron was easily removed from the reaction mixture by a magnetic field, it was reused for ten times without loss in its activity. The ferromagnetic Dacron was better activated at pH 5.0. The particles visualized at scanning electron microscope (SEM) had presented different sizes, varying between 721nm and 100µm. Infra red confirmed immobilization on support, as showed by primary amino peaks at 1640 and 1560 cm-1 . The immobilize enzyme presented Km of 2.32 ± 0.48 mM and optimum temperature of 50°C. Bought presented the same thermal stable of the soluble enzyme and larger enzymatic activity at pH 5.5. β-D-N-acetilglucosaminidase-Dacron ferromagnético showed sensible for some íons as the silver (AgNO3), with loss of activity. The β-D-N acetilglucosaminidase activity for mercury chloride (HgCl2), whom is one of the most toxic substance joined in nature, it was presented activity already diminished at 0,01mM and lost total activity at 4mM, indicating sensitivity for this type of metal. β-D-N-acetilglucosaminidase-ferromagnetic Dacron showed degradative capacity on heparan sulfate, the enzyme still demonstrated degradative capacity on heparan sulphate, suggesting a possible application to produce fractions of this glycosaminoglycan

Polissacarídeos sulfatados de interesse farmacológico no camarão litopenaeus schimitti

Sulfated Polysaccharides with unique chemical structures and important biological activities has been found in a diversity of sea invertebrates. For that, to exist a huger interest on the biotechnology field in the research theses sulfated compounds isolated from sea organisms. Despite the privileged brazilian position for these compounds attainment, there are still a few scientific informations about the isolated substances and their biological activities. A head the displayed, the present work has for objectives, to evaluate the pharmacological properties of the glycosaminoglycans isolated from the sea shrimp Litopenaeus schimitti on homeostasis, blood coagulation, leukocytes migration and platelet/leukocyte adhesion. For this, yhe glycosaminoglycans were extracted from crustacean tissues by proteolysis, fractionation with acetone and later submitted to pharmacological assays. The crustacean tissues showed compounds heparin-like, with anticoagulant activity of 45 IU/mg and 90 IU/mg, respectively. These molecules showed low residual hemorrhagic effects in the tested concentration (100 µg/mL), when compared to unfractionated commercial heparin (UFH). Another dermatan sulfate-like compound, predominately constituted for disulfated disaccharides, was isolated from crustacean abdomen. This compound showed an efficient effect on leukocytes migration inhibition, in the concentration of 15 µg/mL, reducing the cellular infiltration in 65% when compared to the controlled animals. In this same concentration, the DS reduced in 60% the protein concentration of the peritoneal exudates. In the concentration, this compound of 0.5 mg/mL, it was capable to reduce in 40% platelet/leukocytes adhesion. Our data demonstrate that these sulfated polysaccharides isolated from the shrimp L. schimitti will can be used as bioactive compounds, appearing as active principles for pharmacological development, anticoagulants and inflammatory response regulators

Ecologia parasitária de isópodos e biologia reprodutiva em tibiro, Oligoplites spp (OSTEICHTHYES: CARANGIDAE) das águas costeiras de Natal, Rio Grande do norte

Oligoplites palometa (Cuvier) and Oligoplites saurus (Bloch & Schneider) (Osteichthyes: Carangidae) are coastal pelagic fish species. The objective of this study was to investigate the effect of parasitism by isopods and the reproductive biology of the leather jack, Oligoplites spp. A total of 113 individuals of Oligoplites spp (35 of O. saurus and 78 of O. palometa) were captured during the period between January, 2005 and July, 2007 from the coastal waters of Natal, RN. The morphometric and meristic characters were registered; macroscopic analyses were done to collect the isopod parasites and the gonads were observed to verify the reproductive aspects. The crustacean isopod parasite, Cymothoa spinipalpa (Isopoda: Cymothidae) was encountered on the tongue in the oral cavity in both fish hosts. The parasitic indices of C. spinipalpa showed high values of prevalance of 51,4% for O. saurus and 64,1% for O. palometa. The host presented significant correlations between body mass and body length of O. palometa and the number of parasites. Body lengths of female C. spinipalpa were small due to the limited space available in the oral cavity of the host. There was a significant difference in the sex ratio of O. palometa, males outnumbered the females. The isopod parasites showed specificity to the oral cavity of hosts. Macroscopic analyses of gonads showed four stages of gonadal development for both sex of O. palometa: immature, maturing, mature and spent

Diversity of decapod crustaceans at the interface of unconsolidated seabed areas and rocky shores in tropical/subtropical Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Growth of the mangrove crab Ucides cordatus (Brachyura, Ocypodidae)

During monthly samplings between September 1998 and August 2000. 3,660 specimens of Ucides cordatus (Linnaeus, 1763) (2054 males and 1606 females) were obtained and examined for size (CW carapace width) to determine growth-age equations for each sex. This species showed a slower growth, with a marked seasonal oscillation, in females as compared to males, suggesting application of the seasonal and nonseasonal von Bertalanffy growth model, respectively. CW∝ and k constant were closely similar for the two sexes (CW∝ (male) = 90.3 mm: CW∝ (female) = 88.6 mm; k(male) = 0.28; k(female) = 0.26). The age at sexual maturity was estimated to be around 3 years, while the age at legal size (CW = 60 mm) was 3.8 and 4.7 years for males and females, respectively. In the laboratory, juvenile stages did not show differences in growth rates under the same temperature and photoperiod conditions.

SEDIMENT BIOTURBATION POTENTIAL of UCA RAPAX and UCA URUGUAYENSIS AS A RESULT of THEIR FEEDING ACTIVITY

Bioturbation of mangrove sediments by Uca uruguayensis (Nobili, 1901) and U. rapax (Smith, 1870) was compared based on the grain-size composition and organic content in surface sediment around the burrow and feeding pellets in two mangrove zones of the Sao Vicente Estuary, state of São Paulo, Brazil. For each species, 25 burrows with active crabs were selected. All pellets within a 15-cm radius of each burrow were carefully collected; samples of substrate were taken; and the crab occupant was excavated, sexed, and measured for carapace width (CW). The number of spoon-tipped setae on the second maxilliped of each species was estimated; U. uruguayensis showed more of these setae than U. rapax. For both species, the sediment post-processed by feeding activity (feeding pellets) showed a similar increase of coarser fractions and a smaller organic content. However. U. uruguayensis was more efficient in removing organic matter (88.1%) from the sediment than U. rapax (37.5%). These results suggest that different numbers of spoon-tipped setae on the second maxillipeds of the fiddler crabs do not affect the potential for grain-size selection, but result in differing abilities to remove organic matter from the sediment.

Effect of density on population development in the Amazon River prawn Macrobrachium amazonicum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fauna parasitária de peixes oriundos de pesque-pagues do município de Franca, São Paulo, Brasil. II. Metazoários

Metazoan fauna from cultivated fishes (N = 433) in two feefishing farm of Franca, State of São Paulo, Brazil, was studied during a period of April 1997 through March 1999. Specimens of piauçú Leporinus macrocephalus Garavello & Britski, 1988 (Anostomidae), pacu Piaractus mesopotamicus Holmberg, 1887 (Characidae), carp Cyprinus carpio Linnaeus, 1758 (Cyprinidae), Tillapia rendalli Boulenger, 1896 (Cichlidae), nile-tilapia Oreochromis niloticus Linnaeus, 1758 (Cichlidae), matrinxã Brycon cephalus Günther, 1869 (Characidae) and tambacu hybrid (male of P. mesopotamicus x female of Colossoma macropomum Cuvier, 1818) were collected. A total of 29.3% was parasitized by monogenean, 9.7% with crustacean (copepodits and adults of Lernaea cyprinacea Linnaeus, 1758 and Argulus sp.) and 1.6% with myxosporean (Henneguya piaractus Martins & Souza, 1997 and Myxobolus colossomatis Moinar & Békési, 1993).The most susceptible species to myxosporean infestation was P. mesopotamicus. L. macrocephalus and P. mesopotamicus that showed a higher susceptibility to Argulus sp infestation. It was not observed significant difference (P > 0.05) in the seasonal variation of L. cyprinacea infection of all species. L. macrocephalus, P. mesopotamicus and tambacu showed a great number of P. pillulare with no significant difference (P > 0.05) between them. Monogenean infections were observed in all examined species, without significant difference (P > 0.05) in the seasonal variation. Nevertheless, occurrence of monogenean was higher in P. mesopotamicus and lower in Nile-tilapia. By the other hand, parasites number collected from P. mesopotamicus was lower in the winter. Copepodits of L. cyprinacea were found in the gills of P. mesopotamicus, tambacu and L. macrocephalus. However, adults of L. cyprinacea were observed only in the P. mesopotamicus and tambacu.