Elektromagnetsko polje na frekvenciji mobilnih telefona (900 MHz) izaziva stres i modifikacije DNA u gujavici Eisenia fetida

Eisenia fetida earthworms were exposed to electromagnetic field (EMF) at a mobile phone frequency (900 MHz) and at field levels ranging from 10 to 120 V m-1 for a period of two hours (corresponding to specific absorption rates ranging from 0.13 to 9.33 mW kg-1). Potential effects of longer exposure (four hours), field modulation, and a recovery period of 24 h after two hours of exposure were addressed at the field level of 23 V m-1. All exposure treatments induced significant DNA modifications as assessed by a quantitative random amplified polymorphic DNA-PCR. Even after 24 h of recovery following a two hour-exposure, the number of probe hybridisation sites displayed a significant two-fold decrease as compared to untreated control earthworms, implying a loss of hybridisation sites and a persistent genotoxic effect of EMF. Expression of genes involved in the response to general stress (HSP70 encoding the 70 kDa heat shock protein, and MEKK1 involved in signal transduction), oxidative stress (CAT, encoding catalase), and chemical and immune defence (LYS, encoding lysenin, and MYD, encoding a myeloid differentiation factor) were up-regulated after exposure to 10 and modulated 23 V m-1 field levels. Western blots showing an increased quantity of HSP70 and MTCO1 proteins confirmed this stress response. HSP70 and LYS genes were up-regulated after 24 h of recovery following a two hour-exposure, meaning that the effect of EMF exposure lasted for hours.U ovom istraživanju gujavice vrste Eisenia fetida bile su izložene elektromagnetskom polju (EMP) na frekvenciji mobilnih telefona (900 MHz) te poljima jačine 10 do 120 V m-1 u dvosatnom razdoblju (što odgovara specifičnim ratama apsorpcije od 0,13 do 9,33 mW kg-1). Utjecaj dužeg izlaganja (4 sata), modulacije polja te vrijeme oporavka od 24 sata nakon dva sata izlaganja proučavan je pri jačini polja od 23 V m-1. Metoda kvantitativne nasumično umnožene polimorfne DNA (engl. quantitative random amplified polymorphic DNA – qRAPD) otkrila je značajne modifikacije DNA na svim proučavanim tretmanima. Čak i nakon 24-satnog oporavka broj hibridizacijskih mjesta bio je dvostruko manji u odnosu na broj zabilježen u kontrolnim gujavicama, što upozorava na gubitak hibridizacijskih mjesta i na dugoročan utjecaj EMP-a. Ekspresija gena uključenih u odgovor na stres (HSP70: kodira za 70kDa heat shock protein i MEKK1: uključen u provođenje signala), oksidacijski stres (CAT: kodira za katalazu) te kemijsku i imunosnu obranu (LYS: kodira za lysenin i MYD: kodira za faktor mijeloidne diferencijacije) bila je povišena nakon izlaganja polju jačine 10 V m-1 te moduliranome polju jačine 23 V m-1. Western blot analiza potvrdila je odgovor na stres detekcijom povišene količine HSP70 i MTCO1 proteina. HSP70 i LYS geni imali su povišenu ekspresiju i nakon razdoblja oporavka, što upućuje na dugotrajan utjecaj EMP-a

Invasion biology in non-free-living species: interactions between abiotic (climatic) and biotic (host availability) factors in geographical space in crayfish commensals (Ostracoda, Entocytheridae)

In invasion processes, both abiotic and biotic factors are considered essential, but the latter are usually disregarded when modeling the potential spread of exotic species. In the framework of set theory, interactions between biotic (B), abiotic (A), and movement-related (M) factors in the geographical space can be hypothesized with BAM diagrams and tested using ecological niche models (ENMs) to estimate A and B areas. The main aim of our survey was to evaluate the interactions between abiotic (climatic) and biotic (host availability) factors in geographical space for exotic symbionts (i.e., non-free-living species), using ENM techniques combined with a BAM framework and using exotic Entocytheridae (Ostracoda) found in Europe as model organisms. We carried out an extensive survey to evaluate the distribution of entocytherids hosted by crayfish in Europe by checking 94 European localities and 12 crayfish species. Both exotic entocytherid species found, Ankylocythere sinuosa and Uncinocythere occidentalis, were widely distributed in W Europe living on the exotic crayfish species Procambarus clarkii and Pacifastacus leniusculus, respectively. No entocytherids were observed in the remaining crayfish species. The suitable area for A. sinuosa was mainly restricted by its own limitations to minimum temperatures in W and N Europe and precipitation seasonality in circum-Mediterranean areas. Uncinocythere occidentalis was mostly restricted by host availability in circum-Mediterranean regions due to limitations of P. leniusculus to higher precipitation seasonality and maximum temperatures. The combination of ENMs with set theory allows studying the invasive biology of symbionts and provides clues about biogeographic barriers due to abiotic or biotic factors limiting the expansion of the symbiont in different regions of the invasive range. The relative importance of abiotic and biotic factors on geographical space can then be assessed and applied in conservation plans. This approach can also be implemented in other systems where the target species is closely interacting with other taxa

Kavezno izlaganje lubina (Dicentrarchus labrax) u procjeni genotoksičnog utjecaja onečišćenja

Genotoxic effects are often the earliest signs of pollution-related environmental disturbance. In this study, we used the comet assay and micronucleus test to assess DNA damage in the erythrocytes of the European sea bass (Dicentrarchus labrax) exposed to environmental pollution in situ. Fish were collected from a fi sh farm in the Trogir Bay and their cages placed at an unpolluted reference site Šolta (Nečujam Bay) and a polluted site Vranjic (Kaštela Bay) for four weeks. A group of fi sh which remained at the fi sh farm Trogir Bay were used as the second control group. Fish exposed at the Vranjic site showed a signifi cantly higher erythrocyte DNA damage, measured by the comet assay, than either control group. Micronucleus induction showed a similar gradient of DNA damage, but did not reach statistical signifi cance. Our results show that cage exposure of a marine fi sh D. labrax can be useful in environmental biomonitoring and confi rm the comet assay as a suitable tool for detecting pollution-related genotoxicity.Genotoksični učinak često je jedan od najranijih pokazatelja štetnog djelovanja onečišćenja okoliša. U ovom radu procijenjeno je oštećenje DNA u eritrocitima lubina (Dicentrarchus labrax) izloženima okolišnom onečišćenju s pomoću komet-testa i mikronukleus-testa. Lubini su prikupljeni na ribogojilištu i kavezno izloženi u periodu od četiri tjedna na dvije postaje različitog stupnja onečišćenja na jadranskoj obali: na kontrolnoj postaji Šolta (zaljev Nečujam) i na onečišćenoj postaji Vranjic (Kaštelanski zaljev). Zasebna skupina lubina skupljena na ribogojilištu poslužila je kao druga kontrola. Rezultati komet-testa pokazali su statistički značajan porast oštećenja DNA na postaji Vranjic u usporedbi s obje kontrolne postaje. Rezultati mikronukleus-testa pokazali su sličan gradijent onečišćenja, iako nisu dosegli statističku značajnost. Ovi rezultati upućuju na primjenjivost kaveznog izlaganja lubina D. labrax u biomonitoringu vodenog okoliša te potvrđuju korisnost komet-testa kao prikladne metode za detekciju genotoksičnog utjecaja onečišćenja

COVID-19 Lockdowns—Effect on Concentration of Pharmaceuticals and Illicit Drugs in Two Major Croatian Rivers

In order to prevent the spread of COVID-19, contingency measures in the form of lockdowns were implemented all over the world, including in Croatia. The aim of this study was to detect if those severe, imposed restrictions of social interactions reflected on the water quality of rivers receiving wastewaters from urban areas. A total of 18 different pharmaceuticals (PhACs) and illicit drugs (IDrgs), as well as their metabolites, were measured for 16 months (January 2020–April 2021) in 12 different locations at in the Sava and Drava Rivers, Croatia, using UHPLC coupled to LCMS. This period encompassed two major Covid lockdowns (March–May 2020 and October 2020–March 2021). Several PhACs more than halved in river water mass flow during the lockdowns. The results of this study confirm that Covid lockdowns caused lower cumulative concentrations and mass flow of measured PhACs/IDrgs in the Sava and Drava Rivers. This was not influenced by the increased use of drugs for the treatment of the COVID-19, like antibiotics and steroidal anti-inflammatory drugs. The decreases in measured PhACs/IDrgs concentrations and mass flows were more pronounced during the first lockdown, which was stricter than the second

Using a Battery of Bioassays to Assess the Toxicity of Wastewater Treatment Plant Effluents in Industrial Parks

Bioassays, as an addition to physico-chemical water quality evaluation, can provide information on the toxic effects of pollutants present in the water. In this study, a broad evaluation of environmental health risks from industrial wastewater along the Yangtze River, China, was conducted using a battery of bioassays. Toxicity tests showed that the wastewater treatment processes were effective at lowering acetylcholinesterase (AChE) inhibition, HepG2 cells’ cytotoxicity, the estrogenic effect in T47D-Kbluc cells, DNA damage of Euglena gracilis and the mutagenicity of Salmonella typhimurium in the analyzed wastewater samples. Polycyclic aromatic hydrocarbons (PAHs) were identified as potential major toxic chemicals of concern in the wastewater samples of W, J and T wastewater treatment plants; thus, the potential harm of PAHs to aquatic organisms has been investigated. Based on the health risk assessment model, the risk index of wastewater from the industrial parks along the Yangtze River was below one, indicating that the PAHs were less harmful to human health through skin contact or respiratory exposure. Overall, the biological toxicity tests used in this study provide a good basis for the health risk assessment of industrial wastewater and a scientific reference for the optimization and operation of the treatment process

Recent changes in distribution pattern of freshwater crayfish in Croatia − threats and perspectives

Analysis of Croatian freshwater crayfish populations, aiming to gather new distributional data and complement previous surveys (2005–2011), was performed during 2014–2016, within the frame of Natura 2000 Integration Project. The research included different waterbodies across the whole of Croatia, harbouring both indigenous and non-indigenous crayfish species (ICS and NICS, respectively). Field work was conducted in 117 grid squares with dimensions 10 × 10 km, and up to four waterbodies were surveyed per grid square, making a total of 450 studied sites. Out of those, crayfish were not recorded in 368 sites. In the sites with crayfish presence (i.e., 82), the most frequently observed ICS was Astacus astacus (registered in 33 sites) followed by Austropotamobius pallipes (16 sites), Austropotamobius torrentium (12 sites), and Astacus leptodactylus (5 sites). Concerning NICS, the majority of records were for Orconectes limosus (13 sites), followed by Pacifastacus leniusculus (2 sites), whereas Procambarus fallax f. virginalis was registered in only one locality. Comparisons of obtained data with those from previous surveys showed that NICS are progressively spreading and displacing ICS, as recorded for A. leptodactylus that was almost completely displaced by O. limosus in waterbodies of the east Croatia. Existing ICS populations are under growing anthropogenic preassure, frequently fragmented and isolated. Moreover, this survey showed that in the last decade 55% of A. astacus and 67% of A. pallipes populations disappeared mainly as a consequence of anthropogenic influence onto their habitats. Further monitoring and conservation actions for ICS should be urgently applied to mitigate negative impacts of both NICS and anthropogenic influence

Recent changes in distribution pattern of freshwater crayfish in Croatia − threats and perspectives

Analysis of Croatian freshwater crayfish populations, aiming to gather new distributional data and complement previous surveys (2005–2011), was performed during 2014–2016, within the frame of Natura 2000 Integration Project. The research included different waterbodies across the whole of Croatia, harbouring both indigenous and non-indigenous crayfish species (ICS and NICS, respectively). Field work was conducted in 117 grid squares with dimensions 10 × 10 km, and up to four waterbodies were surveyed per grid square, making a total of 450 studied sites. Out of those, crayfish were not recorded in 368 sites. In the sites with crayfish presence (i.e., 82), the most frequently observed ICS was Astacus astacus (registered in 33 sites) followed by Austropotamobius pallipes (16 sites), Austropotamobius torrentium (12 sites), and Astacus leptodactylus (5 sites). Concerning NICS, the majority of records were for Orconectes limosus (13 sites), followed by Pacifastacus leniusculus (2 sites), whereas Procambarus fallax f. virginalis was registered in only one locality. Comparisons of obtained data with those from previous surveys showed that NICS are progressively spreading and displacing ICS, as recorded for A. leptodactylus that was almost completely displaced by O. limosus in waterbodies of the east Croatia. Existing ICS populations are under growing anthropogenic preassure, frequently fragmented and isolated. Moreover, this survey showed that in the last decade 55% of A. astacus and 67% of A. pallipes populations disappeared mainly as a consequence of anthropogenic influence onto their habitats. Further monitoring and conservation actions for ICS should be urgently applied to mitigate negative impacts of both NICS and anthropogenic influence

New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Abstract Background Austropotamobius torrentium is a freshwater crayfish species native to central and south-eastern Europe, with an intricate evolutionary history and the highest genetic diversity recorded in the northern-central Dinarides (NCD). Its populations are facing declines, both in number and size across its entire range. By extanding current knowledge on the genetic diversity of this species, we aim to assist conservation programmes. Multigene phylogenetic analyses were performed using different divergence time estimates based on mitochondrial and, for the first time, nuclear DNA markers on the largest data set analysed so far. In order to reassess taxonomic relationships within this species we applied several species delimitation methods and studied the meristic characters with the intention of finding features that would clearly separate stone crayfish belonging to different phylogroups. Results Our results confirmed the existence of high genetic diversity within A. torrentium, maintained in divergent phylogroups which have their own evolutionary dynamics. A new phylogroup in the Kordun region belonging to NCD has also been discovered. Due to the incongruence between implemented species delimitation approaches and the lack of any morphological characters conserved within lineages, we are of the opinion that phylogroups recovered on mitochondrial and nuclear DNA are cryptic subspecies and distinct evolutionary significant units. Conclusions Geographically and genetically isolated phylogroups represent the evolutionary legacy of A. torrentium and are highly relevant for conservation due to their evolutionary distinctiveness and restricted distribution