ALBERT

Description: Abstract ETHNOPHARMACOLOGICAL RELEVANCE: The oleoresin of Brazilian Copaifera reticulata is a traditional remedy used for the treatment of skin and urinary tract infections, respiratory diseases, rheumatism, ulcer and tumours; thus, playing an important role in the primary health care of the indigenous population. AIM: As most previous pharmacological tests used the crude oleoresin and only a few studies so far dealt with enriched fractions or pure chemically defined compounds, the aim of this study was to systematically evaluate the antimicrobial and cytotoxic properties of the Copaifera reticulata oleoresin and to assign traditional uses to specific secondary metabolites. MATERIALS AND METHODS: The oleoresin, as well as its neutral and acidic fractions were tested for their activity against six cancer cell lines, two clinically relevant bacterial strains, and two dermatophytes. Both fractions were analysed by GC-MS and UHPLC-ELSD, respectively. The antibacterial acidic phase was further fractionated by preparative chromatography to purify and characterize the compounds responsible for the observed pharmacological effect. RESULTS: Whereas no cytotoxic activity was detected, the crude oleoresin and its acidic fraction showed antibacterial activity against gram-positive bacteria Enterococcus faecium (IC50 values 4.2 and 4.8 µg/mL, respectively) and methicillin-resistant Staphylococcus aureus (MRSA, IC50 values 5.3 and 7.2 µg/mL, respectively). Purification of the acidic fraction of the C. reticulata oleoresin yielded two dicarboxylic diterpene acids and the four main diterpene acids, comprising three different diterpene scaffolds. Interestingly, the activity was not restricted to a particular diterpene-type but rather depended on the compounds' lipophilicity, with the most active constituent showing IC50 values of 1.6 (E. faecium) and 2.5 µg/mL (MRSA), respectively. Furthermore, ent-polyalthic acid, the major diterpenoid, was significantly active against dermatophytes with IC50 values of 6.8 µg/mL (Trichophyton rubrum) and 4.3 µg/mL against (T. mentagrophytes). CONCLUSION: The present study proved the antimicrobial effects of the C. reticulata oleoresin and its diterpenoid constituents, confirming its wide use in folk medicine for the treatment of skin and urinary tract infections. The inhibitory activity of copaiba diterpenoids against dermatophytic fungi as well as the gram-positive bacteria E. faecium and MRSA is being reported for the first time, providing potential lead structures for the treatment of these clinically relevant bacterial strains.

Description: The present study investigated the fine-scale population genetic structure of sympatric asterinid sea stars with contrasting modes of larval development (benthic versus pelagic). Parvulastra exigua lacks a dispersive life phase yet is one of the worlds most widely distributed and abundant sea stars, whereas Meridiastra calcar, a sea star with a dispersive larva, has a more limited regional scale distribution. Populations of P. exigua sampled from tide pools on three adjacent headlands showed significant genetic substructure (mitochodrial DNA control region) at fine spatial scales (tide pools 〈 300 m apart: FST = 0.249, P 〈 0.01; headlands 5–15 km apart: FST = 0.125, P = 0.04). As expected, M. calcar populations sampled from the same headlands did not exhibit significant genetic structuring (FST = 0.029, P = 0.14). The life-history traits of P. exigua, a mixed mating system (selfing + outcrossing), pseudocopulation among closely-related conspecifics, and an entirely benthic life cycle with a philopatric larva, undoubtedly influence its strong genetic structure across fine spatial scales. Localized genetic structure, especially at the very fine-scale of tide pools, would not be detected in the more typical regional scale approaches adopted by most studies of marine invertebrate populations.

Description: The effects of different phases of semidiurnal and spring-neap tidal cycles on phytoplankton and environmental drivers were evaluated in a tidal, freshwater location of a mesotidal estuary (Guadiana estuary, SW Iberia). An Eulerian approach was used and sampling covered different seasons during 2008. Samples were collected during spring and neap tides, at high tide, mid-ebb, low tide and mid-flood. Several physical-chemical variables were measured, as well as phytoplankton abundance and biomass. Salinity was higher at high-tide and suspended particulate matter was higher during spring tides and flood, due to higher vertical mixing and resuspension of bottom sediments. Chlorophyll a concentration during winter and summer neap tides was higher than during spring tides, whilst the abundance of pennate diatoms was higher during winter and Spring spring tides than during neap tides, probably reflecting differences in river discharge. Overall, tidally-induced differences detected in the freshwater tidal reaches of the Guadiana estuary were not as considerable as those observed in the lower estuary. However, the occurrence of tidally-induced variability in some seasons reflects that thorough sampling programs to study estuarine tidal dynamics should be conducted throughout the year. Occasional sampling will not reflect the typical variability of these highly dynamic systems. (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Description: Identification of the limiting nutrient(s) is a requirement for the rational management of eutrophication. Here, we present the first experimental analysis of nutrient limitation of phytoplankton growth and its seasonal variation in the Guadiana estuary (SE Portugal-SW Spain). Ten microcosm experiments were performed during 2005 and 2008, using water samples collected in the freshwater tidal zone of the Guadiana estuary. Nitrate, phosphate and silicate were added in a single pulse, alone and in combinations. Experimental treatments were incubated for 4 days under controlled laboratory conditions. Phytoplankton response to nutrient enrichment was evaluated through changes in biomass (Chla), and abundance of specific phytoplankton groups. Overall, phytoplankton growth seemed to be nitrogen-limited throughout the productive period, especially green algae in 2005 and diatoms in 2008. In the summer 2008, cyanobacteria and the harmful dinoflagellate Kryptoperidinium foliaceum responded to N enrichment in the absence of Si. Indeed, the presence of K. foliaceum was observed for the first time in the freshwater tidal reaches of the Guadiana estuary, where dinoflagellates were usually absent or rare. The significant increase on dinoflagellates and cyanobacteria growth in response to N enrichment in the absence of Si is alarming, because anthropogenic nutrient enrichments usually increase N and P, but not Si. Furthermore, relatively high N concentrations, up to 22 μM, were found to be limiting to phytoplankton growth. These results should therefore be used as a management tool when establishing nutrient criteria and nutrient loading budgets to estuarine waters.

Description: Light is usually the main driver of phytoplankton growth in turbid estuaries, but it has received far less attention than nutrients as a bottom-up factor. This study presents the first experimental analysis of light limitation of phytoplankton growth and production and its seasonal variability in the freshwater tidal reaches of the turbid Guadiana estuary, SE Portugal/SW Spain. Natural phytoplankton communities were exposed to different photosynthetically active radiation (PAR) intensities. Short-term incubations with addition of 14HCO3− were used to estimate photosynthetic parameters and long-term incubations allowed the evaluation of the effects of light on phytoplankton composition and growth. Light limitation of phytoplankton growth occurred throughout the year in the freshwater tidal reaches of the estuary and no photoinhibition was observed at least up to 615 μmol photons m−2 s−1. In the summer, co-limitation by nutrients prevented a positive response of phytoplankton to light enrichment. Diatoms were the most light-limited group, whilst cyanobacteria were the only group acclimated to low-light conditions. Green algae and dinoflagellates responded positively to higher PAR exposures. High saturating irradiances, high light-saturated rates of primary production and low photosynthetic efficiencies suggest that phytoplankton community was not acclimated to the low-light conditions that prevail in the Guadiana estuary.

Description: Nitrate and ammonium are the most important nitrogen sources for phytoplankton growth. Differential utilization of inorganic nitrogenous compounds by phytoplankton has been observed and may have significant impacts on primary productivity at local scales. We used enrichment experiments with natural phytoplankton populations from the freshwater tidal zone of the Guadiana estuary, a coastal ecosystem increasingly subjected to anthropogenic influences, to study the effects of nitrate and ammonium on N-consumption and phytoplankton growth. In addition, we used combined additions of nitrate and ammonium to understand the inhibitory effect of ammonium over nitrate uptake. Ammonium concentrations in the freshwater tidal reaches of the Guadiana estuary throughout the sampling period were too low to exert an inhibitory effect on nitrate uptake or a toxic effect on phytoplankton growth. Nitrate was clearly the main nitrogen source for phytoplankton at the study site. Overall, nitrate seemed to become limiting at concentrations lower than 20 mu M and N-limitation was particularly significant during summer. A trend of decreasing nitrate uptake with increasing ammonium concentrations and uptake suggested an overall preference for ammonium. However, preference for ammonium was group-specific, and it was observed mainly in green algae and cyanobacteria. In fact, cyanobacteria relied only on ammonium as their N-source. On the contrary, diatoms preferred nitrate, and did not respond to ammonium additions. The increasing eutrophication in the Guadiana estuary and particularly increased inputs of nitrogen as ammonium due to urban waste effluents may result in a shift in phytoplankton community composition, towards a dominance of cyanobacteria and green algae.

Description: To achieve effective visual camouflage, prey organisms must combine cryptic coloration with the appropriate posture and behaviour to render them difficult to be detected or recognized. Body patterning has been studied in various taxa, yet body postures and their implementation on different backgrounds have seldom been studied experimentally. Here, we provide the first experimental evidence that cuttlefish (Sepia officinalis), masters of rapid adaptive camouflage, use visual cues from adjacent visual stimuli to control arm postures. Cuttlefish were presented with a square wave stimulus (period = 0.47 cm; black and white stripes) that was angled 0°, 45° or 90° relative to the animals' horizontal body axis. Cuttlefish positioned their arms parallel, obliquely or transversely to their body axis according to the orientation of the stripes. These experimental results corroborate our field observations of cuttlefish camouflage behaviour in which flexible, precise arm posture is often tailored to match nearby objects. By relating the cuttlefishes' visual perception of backgrounds to their versatile postural behaviour, our results highlight yet another of the many flexible and adaptive anti-predator tactics adopted by cephalopods.

Description: The potential for aquaculture of the cephalopod species Octopus vulgaris is evaluated, taking into consideration biological and physiological characteristics, as well as some economic and marketing aspects, which may be relevant for the future development of octopus farming. O. vulgaris, a widespread, strictly marine species meets many of the requirements to be considered as a candidate for industrial culture: easy adaptation to captivity conditions, high growth rate, acceptance of low-value natural foods, high reproductive rate and high market price. The life cycle from eclosion of eggs to settlement or beginning of the benthonic adult phase is not commercially viable, but the published results from laboratory and pilot scales are promising. Comments are also made on general research lines needed to improve the use of octopus as farmed species in the future.

Description: Water quality and quantity problems in the Guadiana estuary due to a recently built dam have been predicted, including an enhancement of cyanobacteria blooms. The main goal of this work was thus to describe the present phytoplankton dynamics in relation to its environmental drivers and to evaluate the effects of damming on phytoplankton in the Guadiana estuary. Sampling campaigns were conducted during 2007–2009 in 4 locations of the Guadiana estuary, covering the salinity gradient. Phytoplankton-related and physical–chemical variables were analyzed. Throughout our study, light availability was mainly controlled by suspended sediments and it was much lower than saturating intensities described for phytoplankton growth. Therefore, light was probably limiting to phytoplankton growth throughout the year, especially in the middle and upper estuarine zones. Nitrogen limitation of phytoplankton growth occurred occasionally throughout the study period, especially during spring and summer. Overall, light and nutrient availability were mainly controlled by river flow; anthropogenic sources of nutrients to the estuary were negligible. Phytoplankton showed a unimodal cycle with biomass maximum in late spring/early summer, and the typical seasonal succession of freshwater phytoplankton (diatoms, green algae, cyanobacteria) was observed. Diatoms were the main component of the phytoplankton community and their variability closely followed nitrate and river flow variability. The relative abundance of the main phytoplankton groups changed in relation to the period before dam construction, with a decrease on cyanobacteria contribution to total abundance. The environmental perturbation induced by dam construction has now stabilized and resulted in an overall decrease in nutrient concentrations, an increase in light availability and a decrease in cyanobacteria abundance. Highlights: ► Dam construction is expected to significantly alter estuarine dynamics. ► The effects of dam construction on phytoplankton were analyzed. ► Light and nutrient availability were mainly controlled by river flow. ► No changes in phytoplankton composition were observed. ► Cyanobacteria abundance decreased after dam construction.