Examples of high performance liquid chromatography (HPLC) application in marine ecology studies in the northern Adriatic

Photosynthetic pigments have proved to be useful biomarkers of the abundance, composition and physiological status of the phytoplankton biomass in the marine environment. Using HPLC pigment analysis, we determined phytoplankton community structure in three different marine environments: in the area of a fish farm, in the area of sewage outlets, and in the mucilaginous aggregates. At the reference site we observed seasonal changes with prevalence of fucoxanthin-containing phytoplankton (i.e. diatoms) during winter/spring and autumn. In the fish farm area the concentration of chlorophyll a degradation products was higher, whereas in the locally enriched environment of sewage outlets we observed only small changes in taxonomic composition and phytoplankton biomass. The impact of season is more expressed than the impact of sewage discharge. With the use of HPLC pigment analysis we determined the development of phytoplankton community in different stages of mucilage aggregates. Phytoplankton biomass was composed primarily of diatoms, and as the aggregates aged, diatoms increased in the relative biomass. Our examples have proven the usefulness and suitability of HPLC pigment analysis in marine ecology studies

Project BALMAS newsletter, No. 1, December, 2014

Dear reader, welcome to the BALMAS project newsletter! Transnational cooperation in Adriatic region under the BALMAS project: Ballast Water Management System for Adriatic Sea Protection became fully operational with the start of the BALMAS project in November 2013. The BALMAS project integrates all necessary activities to enable a long-term, environmentally efficient, and financially and maritime trans-port sustainable implementation of ballast water management measures in the Adriatic Sea. In November we marked the first birthday of the BALMAS project. We have passed successfully several milestones, some of which are highlighted in this issue. BALMAS project Partners have been fully involved in a number of project activities, such as sampling in selected ports, attendance at numerous workshops and three regular meetings, and appearances in the media where ongoing and future work has been presented. We established also BALMAS Knowledge centre for the purpose of presenting our interim outputs and results to all interested readers. With all performed activities in the first year, the BALMAS project infrastructure is now completely in place, and we are well positioned to achieve planned objectives within the project time framework. Leon Gosar* *BALMAS project Coordinator for the Lead Partner, the Institute for Water of the Re-public of Sloveni

Spektralni odraz pigmenata i procesi u južnom Jadranu - projekt MEDUZA

During two cruises of the MEDUZA experiment conducted in July 2003 and May 2005 various data were obtained over the deepest part of the southern Adriatic Pit. Temperature, salinity, light transmission and fluorescence were measured in July 2003. During May 2005, except CTD measurements the samples were taken for laboratory determination of phytoplankton pigments with HPLC technique. During both cruises irradiance and radiance were measured at 14 wavelengths in the range 340-715 nm with optical probe to about 100 m depth. Bad weather conditions during both cruises caused interruption of measurements that continued after 24 to 48 hours. Stormy conditions have resulted in a slightly different structure of the water column. In the periods of measurements recorded were diurnal rhythms in a series of parameters, caused by solar radiation cycles in the surface layers as well as fluctuations in the deeper layers under the influence of the internal tide. Meteorological conditions and changes in sea level are also described. The analysis included the principal component and correlation analysis between the measured and derived parameters in order to establish a link between the in-situ pigments and multispectral data. During both cruises diurnal vertical oscillations were observed in biotic and abiotic parameters. Chlorophyll pigments prevailed over other pigments preventing other pigments to be distinguished spectrally. Regression models for chlorophyll a were established from radiance and reflectance ratios.Tijekom nekoliko krstarenja u okviru Meduza eksperimenta obavljenih u srpnju 2003 i svibnju 2005 (http://jadran.izor.hr/meduza/meduza_g.htm), pribavljeni su raznovrsni podaci iznad najdubljeg dijela južnog Jadrana. Temperatura, salinitet, propusnost za svjetlost i fluorescencija mjereni su u srpnju 2003. Tijekom svibnja 2005., osim CTD mjerenja uzeti su uzorci za laboratorijsko određivanje pigmenata fitoplanktona. Tijekom oba krstarenja mjereno je zračenje na 14 valnih duljina u rasponu 340-715nm optičkom sondom do oko 100m dubine. Loši vremenski uvjeti tijekom oba krstarenja uzrokovali su prekid mjerenja koja se nastavljaju nakon 24 odnosno 48 sati. Olujni uvjeti rezultirali su nešto drugačijom strukturom vodenog stupca. U razdobljima mjerenja zabilježeni su dnevni ritmovi u nizu parametara, uzrokovani ciklusima sunčeva zračenja u površinskim slojevima kao i oscilacije u dubljim slojevima zbog unutarnje plime. Meteorološki uvjeti i promjene razine mora su također diskutirani. Analiza je obuhvatila i korelacije između izvedenih i mjerenih parametara s ciljem da uspostavi veza između koncentracija mjerenog klorofila i multispektralnih podataka. Tijekom oba krstarenja opažene su dnevne vertikalne oscilacije biotskih i abiotskih pametera. Klorofilni su pigmenti prevladavali onemogućujući spektralno prepoznavanje ostalih pigmenata. Utvrđeni su regresivni modeli za klorofil a na osnovi mjerenja radijanca i reflektancija

Dinoflagellate resting cysts from surface sediments of the Adriatic Ports: distribution and potential spreading patterns

The ability of microalgae to preserve viable in coastal sediments as resting forms provides a reservoir of biodiversity and a useful tool to determine species spreadings. This study represents the first port baseline survey on dinoflagellate cysts, investigated in nine Adriatic ports during a cross border project. 40 dinoflagellate taxa were detected. The assemblages resulted in all ports dominated by Lingulodinium polyedra and Alexandrium minutum/affine/tamutum group. General separation to the western and eastern side of the Adriatic regarding cysts assemblage composition, partially abundance, was observed. Seven taxa were detected as non-indigenous species for the Adriatic. Two taxa are included in the list of harmful aquatic organisms, indicating the potential threat of ballast waters in the Adriatic. Potential spreading of taxa by general circulation and ballast waters, intra- and extra-Adriatic was investigated. The entering in to force of the ballast waters management regulations should enhance prospects to minimize future harmful impacts

Mixtures of Chemical Pollutants at European Legislation Safety Concentrations: How Safe Are They?

The risk posed by complex chemical mixtures in the environment to wildlife and humans is increasingly debated, but has been rarely tested under environmentally relevant scenarios. To address this issue, two mixtures of 14 or 19 substances of concern (pesticides, pharmaceuticals, heavy metals, polyaromatic hydrocarbons, a surfactant, and a plasticizer), each present at its safety limit concentration imposed by the European legislation, were prepared and tested for their toxic effects. The effects of the mixtures were assessed in 35 bioassays, based on 11 organisms representing different trophic levels. A consortium of 16 laboratories was involved in performing the bioassays. The mixtures elicited quantifiable toxic effects on some of the test systems employed, including i) changes in marine microbial composition, ii) microalgae toxicity, iii) immobilization in the crustacean Daphnia magna, iv) fish embryo toxicity, v) impaired frog embryo development, and vi) increased expression on oxidative stress-linked reporter genes. Estrogenic activity close to regulatory safety limit concentrations was uncovered by receptor-binding assays. The results highlight the need of precautionary actions on the assessment of chemical mixtures even in cases where individual toxicants are present at seemingly harmless concentration

Mixtures of chemical pollutants at European legislation safety concentrations: how safe are they?

The risk posed by complex chemical mixtures in the environment to wildlife and humans is increasingly debated, but has been rarely tested under environmentally relevant scenarios. To address this issue, two mixtures of 14 or 19 substances of concern (pesticides, pharmaceuticals, heavy metals, polyaromatic hydrocarbons, a surfactant, and a plasticizer), each present at its safety limit concentration imposed by the European legislation, were prepared and tested for their toxic effects. The effects of the mixtures were assessed in 35 bioassays, based on 11 organisms representing different trophic levels. A consortium of 16 laboratories was involved in performing the bioassays. The mixtures elicited quantifiable toxic effects on some of the test systems employed, including i) changes in marine microbial composition, ii) microalgae toxicity, iii) immobilization in the crustacean Daphnia magna, iv) fish embryo toxicity, v) impaired frog embryo development, and vi) increased expression on oxidative stress-linked reporter genes. Estrogenic activity close to regulatory safety limit concentrations was uncovered by receptor-binding assays. The results highlight the need of precautionary actions on the assessment of chemical mixtures even in cases where individual toxicants are present at seemingly harmless concentrations

Examples of high performance liquid chromatography (HPLC) application in marine ecology studies in the northern Adriatic

Fotosintezna barvila so se izkazala kot dobri kazalci abundance, sestave in fiziološkega stanja fitoplanktonske biomase v morskem okolju. Z uporabo HPLC-analize barvil smo določili strukturo fitoplanktonske združbe v treh različnih morskih okoljih: na območju ribogojnic, na območju komunalnih podvodnih izpustov in v sluzastih makroagregatih. Na referenčni postaji smo izmerili sezonske spremembe fitoplanktonske združbe s prevlado fukoksantin-vsebujočega fitoplanktona (t.j. diatomej) v zimsko-spomladanskem in jesenskem obdobju. Na območju ribogojnic smo izmerili višje koncentracije razgradnih produktov klorofila a, na lokalno obogatenem območju komunalnih izpustov pa smo opazili le manjše spremembe v taksonomski sestavi in biomasi fitoplanktona. Vpliv sezone je bolj izražen kakor vpliv podvodnih komunalnih izpustov. Z uporabo HPLC-analize barvil smo določili razvoj fitoplanktonske združbe v posameznih razvojnih stadijih sluzastih makroagregatov. V fitoplanktonski združbi sluzastih makroagregatov so prevladovale diatomeje in njihov delež k relativni biomasi je s starostjo agregatov še naraščal. Navedeni primeri kažejo na uporabnost in primernost HPLC-analize barvil v študijah morske ekologije.Photosynthetic pigments have proved to be useful biomarkers of the abundance, composition and physiological status of the phytoplankton biomass in the marine environment. Using HPLC pigment analysis, we determined phytoplankton community structure in three different marine environments: in the area of a fish farm, in the area of sewage outlets, and in the mucilaginous aggregates. At the reference site we observed seasonal changes with prevalence of fucoxanthin-containing phytoplankton (i.e. diatoms) during winter/spring and autumn. In the fish farm area the concentration of chlorophyll a degradation products was higher, whereas in the locally enriched environment of sewage outlets we observed only small changes in taxonomic composition and phytoplankton biomass. The impact of season is more expressed than the impact of sewage discharge. With the use of HPLC pigment analysis we determined the development of phytoplankton community in different stages of mucilage aggregates. Phytoplankton biomass was composed primarily of diatoms, and as the aggregates aged, diatoms increased in the relative biomass. Our examples have proven the usefulness and suitability of HPLC pigment analysis in marine ecology studies