Validation of an analytical methodology for the quantitative analysis of petroleum hydrocarbons in marine sediment samples

This work describes a validation of an analytical procedure for the analysis of petroleum hydrocarbons in marine sediment samples. The proposed protocol is able to measure n-alkanes and polycyclic aromatic hydrocarbons (PAH) in samples at concentrations as low as 30 ng/g, with a precision better than 15% for most of analytes. The extraction efficiency of fortified sediments varied from 65.1 to 105.6% and 59.7 to 97.8%, for n-alkanes and PAH in the ranges: C16 - C32 and fluoranthene - benzo(a)pyrene, respectively. The analytical protocol was applied to determine petroleum hydrocarbons in sediments collected from a marine coastal zone.

Austin, dehydroaustin and other metabolites from Penicillium brasilianum

A culture of P. brasilianum, isolated from soil collected at the Serra do Cipó National Park, in Minas Gerais State (Brazil), was grown for 25 days on a dextrose-peptone-salts medium. The corresponding ethylacetate extract was column chromatographed and four compounds were isolated: austin, dehydroaustin, D-mannitol and penicillic acid. This is, in the best of our knowledge, the first time that the meroterpenes austin and dehydroaustin have been isolated from this species. Activity of the extract and isolated compounds was tested against six bacteria and for acetylcholinesterase inhibition. Penicillic acid showed high activity in both tests.

An antimicrobial alkaloid and other metabolites produced by Penicillium sp. An endophytic fungus isolated from Mauritia flexuosa L. f.

The alkaloid glandicoline B (1) and six other compounds: ergosterol (2), brassicasterol (3), ergosterol peroxide (4), cerevisterol (5), mannitol (6) and 1-O-α-D-glucopyranoside (7) were isolated from Penicillium sp. strain PBR.2.2.2, a fungus from Mauritia flexuosa roots. The structures of the isolated metabolites were established by spectral analysis. MeOH extract of the fungal mycelium at 500 µg mL-1 exhibited antimicrobial activity against Staphylococcus aureus and the compound 1 at 100 µg mL-1 was active against S. aureus, Micrococcus luteus and Escherichia coli. The relationship between the bioactive properties of the fungus PBR.2.2.2 and those achieved for glandicoline B, as well the potential of this substance as bactericide is discussed.

Antiparasitic bromotyrosine derivatives from the Caribbean marine sponge Aiolochroia crassa

Six bromotyrosine-derived compounds were isolated from the Caribbean marine sponge Aiolochroia crassa: 3-bromo-5-hydroxy-O-methyltyrosine (1), 3-bromo-N,N,N-trimethyltyrosinium (2), 3-bromo-N,N,N,O-tetramethyltyrosinium (3), 3,5-dibromo-N,N,N-trimethyltyrosinium (4), 3,5-dibromo-N,N,N,O-tetramethyltyrosinium (5), and aeroplysinin-1 (6). Structural determination was performed using NMR, MS and comparison with literature data. All isolated compounds were screened for their in vitro activity against Leishmania panamensis, Plasmodium falciparum and Trypanosoma cruzi. Compound 4 showed selective antiparasitic activity against Leishmania and Plasmodium parasites. This is the first report of compounds 1, 4 and 5 in the sponge A. crassa and the first biological activity reports for compounds 2-4. This work shows that bromotyrosines are potential antiparasitic agents.

Seasonal variation of the major secondary metabolites present in the extract of Eremanthus mattogrossensis Less (Asteraceae: Vernonieae) leaves

The species Eremanthus mattogrossensis, known as "veludo do cerrado" (cerrado velvet), is native to the Brazilian Cerrado. Because the amount of metabolites present in plants may be influenced by biological and environmental factors, here we conducted an HPLC-DAD-MS/MS investigation of the metabolite concentrations found in the MeOH/H2O extract of the leaves of this species. The main compounds were identified and quantified, and the metabolites were grouped by chemical class (caffeoylquinic acids, flavonoids, and sesquiterpene lactone). Statistical analysis indicated a straight correlation between the quantity of metabolites and seasonality, suggesting that environmental properties elicit important metabolic responses.

Phytotoxic effects of metabolites from Alternaria euphorbiicola against its host plant Euphorbia heterophylla

A bioassay-guided fractionation of culture filtrates of the fungus Alternaria euphorbiicola, a pathogen of the weed Euphorbia heterophylla, led to the isolation of anhydromevalonolactone (1), tyrosol (2), (R)-( - )-mevalonolactone (3), and cycloglycylproline (4). When tested on the punctured leaves of the host plant, these compounds produced bleached lesions with dark brown margins at concentrations as low as 80 µM. When tested on the leaves of other relevant weeds, only cycloglycylproline showed selective activity against E. heterophylla. This is the first report on the isolation of phytotoxins from A. euphorbiicola and on the phytotoxicity of anhydromevalonolactone, (R)-( - )-mevalonolactone, and cycloglycylproline.

Analysis of metabolites from plants of the Swartzia genus using chemical indexes: evolutionary tendencies

The chemical indexes, suggested by Gottlieb et al., have not been used before regarding evolutionary tendency of species in the Swartzia genus. However, the importance of this work encouraged for an analysis of the Swartzia genus using the metabolites isolated from nine species. The analysis, based on calculated chemical indexes, provided an evolutionary tendency for these plants, which correlates with the classification based on morphological analysis.

Antimicrobial activity of extracellular metabolites from antagonistic bacteria isolated from potato (Solanum phureja) crops

Microorganisms for biological control are capable of producing active compounds that inhibit the development of phytopathogens, constituting a promising tool toob tain active principles that could replace synthetic pesticides. This study evaluatedtheability of severalpotentialbiocontrol microorganismsto produce active extracellular metabolites. In vitro antagonistic capability of 50 bacterial isolates from rhizospheric soils of "criolla" potato (Solanum phureja) was tested through dual culture in this plant with different plant pathogenic fungi and bacteria. Isolates that showed significantly higher antagonistic activity were fermented in liquid media and crude extracts from the supernatants had their biological activities assessed by optical density techniques. Inhibitory effecton tested pathogens was observed for concentrations between 0.5% and 1% of crude extracts. There was a correlation between the antimicrobial activity of extracts and the use of nutrient-rich media in bacteria fermentation. Using a bioguided method, a peptidic compound, active against Fusarium oxysporum, was obtained from the 7ANT04 strain (Pyrobaculum sp.). Analysis by nuclear magnetic resonance and liquid chromatography coupled to mass detector evidenced an 11-amino acid compound. Bioinformatic software using raw mass data confirmed the presence of a cyclic peptide conformed by 11 mostly non-standard amino acids.

Design of marine generators for alternative diesel-electric power systems

This study compares different electric propulsion systems. Results of the analysis of all the advantages and disadvantages of the different propulsion systems are given. This thesis estimates possibilities to apply different diesel-electric propulsion concepts for different vessel types. Small and medium size vessel’s power ranges are studied. The optimal delivery system is chosen. This choice is made on the base of detailed study of the concepts, electrical equipment market and comparison of mass, volume and efficiency parameters. In this thesis three marine generators are designed. They are: salient pole synchronous generator and two permanent magnet synchronous generators. Their electrical, dimensional, cost and efficiency parameters are compared. To understand all the benefits diagrams with these parameters are prepared. Possible benefits and money savings are estimated. As the result the advantages, disadvantages and boundary conditions for the permanent magnet synchronous generator application in marine electric-power systems are found out.

Effect of light intensity and growth substratum on plant development and production of secondary metabolites in Cordia curassavica (Jacq.) Roem. & Schult

Cordia curassavica (Jacq.) Roem. & Schult. (Boraginaceae), also referred to as Cordia verbenacea DC, has been traditionally used for medicinal purposes. This study was driven to verify the behavior of the species in similar conditions to its natural environment, such as high light intensity and sandbank soil, and in conditions of low light intensity and fertilized substratum (dystroferric red nitosoil plus earthworm humus). The growth of the plant, the income of leaf crude extracts and, in the alcoholic extract, the number of substances found in thin layer cromatography and the toxicity of the substratum was observed. The results indicated that the growth of the root biomass, stem and leaves in discharge or lower light intensity was similar, but smaller in sandbank soil than in fertilized soil. The relative income of extracts in ether of petroleum and alcohol was larger in high light intensity and fertilized substratum. The light intensity and the substratum type didn't affect the number of substances detected in the alcoholic extract or the toxicity of this extract. Stains corresponding to the rosmarinic acid were only evidenced in some samples of the alcoholic extract, not allowing the verification of the effect of the treatments about its production.

Public-private partnership in marine litter waste management

Marine litter is an international environmental problem that causes considerable costs to coastal communities and the fishing industry. Several international and national treaties and regulations have provisions to marine litter and forbid disposal of waste into the sea. However, none of these regulations state a responsibility for public authorities to recover marine litter from the sea, like they do for marine litter that washes up on public beaches. In a financial evaluation of a value chain for marine litter incineration it was found out that the total costs of waste incineration are approximately 100 ─ 200 % higher than waste fees offered by waste contractors of ports. The high costs of incineration are derived from the high calorific value of marine litter and therefore a high incineration cost for the waste, and long distances between ports that are taking part in a project for marine litter recovery from the sea and an Energy-from-Waste (EfW) facility. This study provides a possible solution to diverting marine litter from landfills to more environmentally sustainable EfW use by using a public-private partnership (PPP) framework. PPP would seem to fit as a suitable cooperative approach for answering problems of current marine litter disposal in theory. In the end it is up to the potential partners of this proposed PPP to decide whether the benefits of cooperation justify the required efforts.

The Sedimentology, Ichnology and Hydrogeochemistry of the Late Miocene, Marginal Marine, Upper Pebas and Nauta Formations, Amazonian Foreland Basin, Peru

This thesis includes detailed sedimentological and ichnological studies on two geological units: the Pebas Formation, with a special focus in its informal upper member, and the Nauta Formation. Both formations were deposited during the Miocene in Northeastern Peruvian Amazonia, in the Amazon retroarc foreland basin. The Pebas and Nauta successions mainly consist of non-consolidated, clastic sedimentary deposits arranged into sand- to mud-dominated heterolithic successions, which can be upward-coarsening to upward-fining. Sediments in both the Pebas and Nauta successions range from mud to fine- to medium-grained sand. The main facies observed were 1) mud-dominated horizontal heterolithic couplets; 2) rooted brownish mud; 3) lenticular, mud-draped, cross-stratified sand; 4) mud- to sand-dominated, inclined heterolithic stratification; 5) sand-dominated horizontal heterolithic couplets; and 6) mud-draped, trough cross-stratified sand. Locally, tidal rhythmites were documented. The facies are interpreted as: 1) muddy, shallow, subaqueous flats/shoals; 2) palaeosols; 3) secondary tidal channels or run-off creeks; 4) tidally influenced point bars; 5) shoreface deposits; and 6) subtidal compound dunes. Thalassinoides-dominated Glossifungites ichnofacies, low-diversity expressions of the Skolithos ichnofacies and depauperate suites consisting of elements common to the Cruziana ichnofacies strongly indicate brackish-water conditions. However, continental trace fossil assemblages, with possible elements common to the Scoyenia ichnofacies, have also been identified. In addition to the palaeoenvironmental study, a local hydrogeochemical characterisation of the Pebas and Nauta formations was also conducted. The geochemistry of the groundwaters reflects the characteristics and the soil geochemistry of the geological formations studied. The Pebas formation has low hardness, acid to neutral waters, whereas the upper Pebas has high hardness, acid to neutral waters. In both units, the arsenic content is locally high. The Nauta formation has low hardness acid groundwaters. A regional review of the Pebas and Nauta formations placed the local observations into a continental perspective and suggests that the whole Pebas-Nauta system was a probably shallow (some tens of metres at maximum), brackish- to freshwater, tidally-influenced epicontinental embayment with a probable semi-diurnal to mixed tidal regime and a microtidal range, surrounded by continental environments such as forest floors, lagoons, rivers and their flood plains, and lakes.

Identification and application of bile metabolites to assess the exposure of fish to pharmaceuticals in the environment

Thousands of tons of pharmaceuticals are consumed yearly worldwide. Due to the continuous and increasing consumption and their incomplete elimination in wastewater treatment plants (WWTP), pharmaceuticals and their metabolites can be detected in receiving waters, although at low concentrations (ng to low μg/L). As bioactive molecules the presence of pharmaceuticals in the aquatic environment must be considered potentially hazardous for the aquatic organisms. In this thesis, the biotransformation and excretion of pharmaceuticals in fish was studied. The main biotransformation pathways of three anti‐inflammatory drugs, diclofenac, naproxen and ibuprofen, in rainbow trout were glucuronidation and taurine conjugation of the parent compounds and their phase I metabolites. The same metabolites were present in fish bile in aquatic exposures as in fish dosed with intraperitoneal injection. Higher bioconcentration factor in bile (BCFbile) was found for ibuprofen when compared to diclofenac and naproxen. Laboratory exposure studies were followed by a study of uptake of pharmaceuticals in a wild fish population living in lake contaminated with WWTP effluents. Of the analyzed 17 pharmaceuticals and six phase I metabolites, only diclofenac, naproxen and ibuprofen was present in bream and roach bile. It was shown, that diclofenac, naproxen and ibuprofen excreted by the liver can be found in rainbow trout and in two native fish species living in the receiving waters. In the bream and roach bile, the concentrations of diclofenac, naproxen and ibuprofen were roughly 1000 times higher than those found in the lake water, while in the laboratory exposures, the bioconcentration of the compounds and their metabolites in rainbow trout bile were at the same level as in wild fish or an order of magnitude higher. Thus, the parent compounds and their metabolites in fish bile can be used as a reliable biomarker to monitor the exposure of fish to environmental pharmaceuticals present in water receiving discharges from WWTPs.

Detection and characterization of fibropapilloma associated herpesvirus of marine turtles in Rio Grande do Sul, Brazil

Fibropapillomatosis (FP) is a benign tumoral disease that affects sea turtles, hampering movement, sight and feeding, ultimately leading to death. In Brazil, the disease was described for the first time in 1986. Research suggests the involvement of a herpesvirus in association with environmental and genetic factors as causal agents of FP. The objective of the present study was to detect and characterize this herpesvirus in sea turtles living in the coast of state Rio Grande do Sul (RS), Brazil. From October 2008 to July 2010, 14 turtles were observed between the beaches of Torres and Tavares, of which 11 were green turtles (Chelonia mydas) and 3 were loggerhead turtles (Caretta caretta). All turtles were young and mean curved carapace length was 37.71±7.82cm, and varied from 31 to 55cm. Only one green turtle presented a 1cm, papillary, pigmented fibropapilloma. Skin and fibropapilloma samples were analyzed by conventional and real time PCR assays to detect and quantify herpesvirus. All skin samples were negative, though the fibropapilloma specimen was positive in both tests. Viral load was 9,917.04 copies of viral genome per milligram of tissue. The DNA fragment amplified from the fibropapilloma sample was sequenced and allocated in the Atlantic phylogeographic group. This study reports the first molecular characterization of herpesvirus associated with fibropapilloma in turtles from the coast of RS.

The Role of a firm’s mindset in international success - case study of Finnish marine industry firms

This thesis investigates the influence of a firm’s mindset on international success in the context of the Finnish marine industry. The study draws theoretical wisdom from organisational behaviour and strategic management literatures. The research aim set for this study was to identify and categorise existing strategic types based on mindsets found in the marine industry SMEs, and to link the influence of mindsets with success by examining the role of mindsets in a firm’s performance. Mindsets of firms were conceptualised as aggregate collections of perceptions that influence how the surrounding environment is discerned by the members of the firm. Mindsets are idiosyncratic to firms and therefore important firm-specific resources which influence decision-making and can be observed through the strategic behaviour of firms. Qualitative case study method was applied which was further supported by quantitative data on the financial performance of the ten case firms. Taxonomy based on the dimension of mindsets and prediction was developed to demonstrate four ideal types of firms identified within the marine industry. It was found that all of the case firms emphasised adaptation in their strategy while planning was emphasised to a varying degree. Moreover, two different methods of adapting were found; proactive and reactive. Firms which plan in the long-term and adapt proactively constantly investigate whether their plans are synchronous with the realities of the market; by having an open mindset, a firm’s perception of the reality of the market is enabling the firm to develop value creating strategies which are superiorly informed.This finding was supported by the financial data and led to the proposition that having an open mindset and placing a high level of emphasis on prediction may have a positive influence on international success. Also, it was proposed that concentrating only on exploiting business opportunities in the present time and not exploring any addition opportunities can have a negative influence on the firm’s performance, even if the mindset of the firm is open.