Marija Ivković

Assemblage composition and ecological features of aquatic dance flies (Diptera, Empididae) in the Cetina River system, Croatia

Twenty species of aquatic dance flies (Empididae: Clinocerinae and Hemerodromiinae) were recorded from the Cetina River system, Croatia. No previous such research has been done on any Mediterranean river con-cerning aquatic dance flies. Aduit dance flies were sampled at 7 locations with an aspirator and sweep nets between May and September 2005 and in March and April 2006. The most abundant species was Wiedemannia (Wiedeman-nia) bistigma which was present in ali months but only at locations in the lower reach of the Cetina River. Larvae were sampled at 6 sites from different microhabitat types. Altogether 241 quantitative samples were taken at 20 sampling points between August 2004 and August 2005, but dance flies vvere found only in a small number of the samples. The relationships between dance fly assemblage structure and selected environmental variables (water temperature, dissolved oxygen, alkalinity, conductivity and pH) vvere investigated and showed that the most im-portant environmental variable was water temperature. Some species clearly preferred sites where the mean annual water temperature was constant and lower year round, like springs (e.g., Chelifera spp., Wiedemannia (Philolutra) kacanskae, Wiedemannia (Chamaedipsia) ariadne), while other species preferred sites where the mean annual water temperature was higher (e.g., Hemerodromia spp., Wiedemannia (Wiedemannia) tricuspidata, Wiedeman-nia {; ; ; Pseudowiedemannia) lamelatta). Some species did not show any strong preferences for any of the selected parameters (e.g., Wiedemannia (Eucelidia) zetterstedti, Wiedemannia (Wiedemannia) bistigma, Dolichocephala guttata). A significant positive correlation emerged between the amount of prey present in different microhabitat types and the number of dance fly larvae. A significant difference was also found between microhabitat types and total number of dance fly larvae.

Assessment of genotoxic potency of sulfate-rich surface waters on medicinal leech and human leukocytes using different versions of the Comet assay.

The aim of the present study was to investigate how exposure to sulfate-rich surface waters affects the level of primary DNA damage in hemocytes of leech Hirudo medicinalis. Samples of surface water were collected at two sites near a gypsum factory (Knin, Croatia) and two reference sites. In the laboratory, samples were subjected to detailed chemical analysis and used in toxicity testing. For that purpose, previously acclimatized individuals of H. medicinalis were sub-chronically exposed (for 28 days) to tested water samples. Levels of primary DNA damage were evaluated using the alkaline Comet assay in hemocytes collected on days 7, 14, 21 and 28 of exposure and compared with their baseline values. Genotoxic potency of the water sample with the highest sulfate concentration was further evaluated using the alkaline, neutral and hOGG1-modified Comet assay on human peripheral blood leukocytes exposed ex vivo for 30 min. The purpose was to explore which mechanisms are responsible for DNA damage. Chemical analysis revealed that sulfate concentrations in two water samples collected in Mali Kukar Lake (1630 mg/L SO₄) and Kosovčica River (823.3 mg/L SO₄) exceeded the WHO and US EPA defined limits for sulfate in drinking water. Increased levels of metals were found only in the water sample collected in Mali Kukar Lake. However, of the 65 elements analyzed, only nickel and titanium exceed the value legally accepted in Croatia for drinking water. The levels of DNA damage, estimated by the alkaline Comet assay in hemocytes of medicinal leech, increased with the duration of exposure to two sulfate-rich water samples. Since hemocytes responded sensitively to treatment, they could be used for biomonitoring purposes. As observed on treated human peripheral blood leukocytes, all versions of the Comet assay were effective in detecting DNA damage, which was measured in samples with sulfate concentrations equal to or higher than the legally accepted levels for drinking water. Based on the obtained results, it can be assumed that genotoxicity was a consequence both of direct (single- and double-strand DNA breaks) and indirect effects (oxidative damage) caused by the combined effects of all contaminants present in the tested water samples. Our results indicate the need for in situ monitoring and purification of gypsum mine water prior to its release in the natural environment.

Aquatic insects in the Dinarides: identifying hotspots of endemism and species richness shaped by geological and hydrological history using Empididae (Diptera)

We identify local hotspots of elevated species richness and endemicity of aquatic insects in the Dinarides (Balkans) using aquatic Empididae (Diptera) as models and compared observed patterns with historical geological and hydrological changes in the region. We analysed species richness and reciprocal weighted endemicity. Parsimony analysis of endemism was used to generate an historical hypothesis of the relationships between the aquatic Empididae biotas of different regions of local endemism within the Dinarides and the results reconciled with the geological and hydrological history of the region. Hotspots of high endemicity were coincident with areas of elevated species richness and were located primarily in the northwest Dinarides, central Bosnia & Herzegovina, and southeast Montenegro. Parsimony analysis of endemism retrieved three weakly supported area clades comprising areas of southeast Dinarides together with disjunct areas at the western Dinarides, and central Bosnia & Herzegovina together with the disjunct northwest Dinarides. The aquatic Empididae fauna of the northwest Dinarides has been strongly influenced by dispersal from the European Alps, whereas that at the southern end of the range may have been shaped by immigration from ranges further south. In the central Dinarides, progressive fragmentation of the hydrological network through orogenesis, karstification, redistribution of flysch deposits, and saline ingressions may underlie current diversity patterns with populations becoming increasingly isolated and focused within refugia. Hotspots of endemism and diversity are not necessarily coincident with protected areas and further work will be needed if they are to be reconciled with future planning of conservation priorities.

Emergence patterns and ecological associations of some haematophagous blackfly species along an oligotrophic hydrosystem

Emergence patterns of blackflies (Diptera: Simuliidae) were investigated for 2 years at nine sites in the Plitvice Lakes area of Croatia, using pyramid‐type traps. A total of 38 012 specimens representing 10 haematophagous species were collected. The dominant species was Simulium angustipes Edwards, 1915, a vector transmitting avian trypanosomes. Water temperature, alkalinity, conductivity and habitat type represented the factors with the greatest degree of influence on blackfly species composition, structure and distribution along a longitudinal gradient of this oligotrophic karstic hydrosystem. Simulium angustipes was highly associated with lake outlets. The blackfly communities at all sites were dominated by species typical of the rhithral zone, but there was a shift in species composition along a longitudinal gradient from the hypocrenal–epirhithral to the epirhithral–metarhithral zone. Multiple generations per year were detected in S. angustipes, whereas in Simulium costatum Friederichs, 1920, the number of generations differed between sites with constant and those with variable water temperature.

New records of moth flies (Diptera: Psychodidae) from Croatia, with the description of Berdeniella keroveci sp.nov.

Berdeniella keroveci sp. nov. is described based on males collected in Croatia. Current knowledge of the psychodid fauna of Croatia is reviewed and new records of 16 species are presented. Short taxonomic notes on Pericoma pseudocalcilega Krek, 1972, Pneumia balkanica (Krek, 1990), Pneumia mutua (Eaton, 1893), Clogmia albipunctata (Williston, 1893) and Lepiseodina tristis (Meigen, 1810) are given. In total, 33 species of Psychodidae are known to occur in Croatia.

Blind Flight? A New Troglobiotic Orthoclad (Diptera, Chironomidae) from the Lukina Jama - Trojama Cave in Croatia.

The genus Troglocladius Andersen, Baranov et Hagenlund, gen. n. is erected based on T. hajdi Andersen, Baranov et Hagenlund, sp. n. collected at 980 m depth in the Lukina jama—Trojama cave system in Croatia. Morphological features such as pale color, strongly reduced eyes and very long legs make it a typical cave animal. Surprisingly, it has also retained large wings and appears to be capable of flight which would make T. hajdi the first flying troglobiont worldwide, disproving previous beliefs that bats are the only animals capable of flying in complete darkness. Morphologically the new species does not readily fit within any described genus, but shares characteristics with genera both in the tribes “Metriocnemini” and “Orthocladiini”. Bayesian molecular phylogenetic analysis using the markers COI, 18S rDNAs, 28S rDNA, CADI, and CADIV groups it with the genera Tvetenia, Cardiocladius and Eukiefferiella in the tribe “Metriocnemini”. Troglocladius hajdi may be parthenogenetic, as only females were collected. The discovery confirms the position of the Dinaric arch as a highly important hotspot of subterranean biodiversity.

The impact of gypsum mine water: A case study on morphology and DNA integrity in the freshwater invertebrate, Gammarus balcanicus

The aim of our study was to investigate how exposure to heavy metal-rich waters from gypsum mining affects the morphology and levels of primary DNA damage in Gammarus balcanicus. Chemical analysis revealed increased concentrations of metals in water and sediment collected at a site impacted by gypsum mine wastewaters. The specimens also showed elevated total tissue metal levels when compared with the organisms collected at the reference site. The most prominent increase was observed for strontium, followed by iron, nickel, vanadium, aluminium, and manganese. The major pathway of entry for these toxic substances was through the degraded exoskeleton as a consequence of excessive strontium input (unbalanced calcium/strontium ratio) and altered permeability. Disturbed exoskeleton integrity was observed only in individuals collected downstream of the gypsum mine, which was confirmed by electron microscopy. Levels of primary DNA damage were evaluated using the alkaline comet assay in the haemolymph of the specimens.

The hunter-flies of Croatia (Diptera: Muscidae: genus Limnophora Robineau-Desvoidy)

An identification key is given to the eight species of hunter-flies of the genus Limnophora Robineau-Desvoidy (Muscidae) known from Croatia. Four species are new records for Croatia, and one new species, Limnophora croatica sp. nov., is described. To illustrate the relationship of the new species, a key is given to the Mediterranean members of the Limnophora riparia group of species. http://www.zoobank.org/urn:lsid:zoobank.org:pub:6CD4995D-1C16-4910-81F1-AC59A0E1FA34

New records of Muscidae (Diptera) from Mediterranean countries

Abstract New records are provided for Muscidae from four different Mediterranean countries, with new distribution records for species in ten different genera. Seven species are newly recorded for Croatia, four species for Montenegro and one species for Bosnia & Herzegovina. In this paper we give the first confirmation of an aquatic larval stage for Lispocephala brachialis (Rondani, 1877), Lispocephala spuria (Zetterstedt, 1838) and Lispocephala mikii (Strobl, 1893). A first record of the species Coenosia lyneborgi Pont, 1972 since its original description is also provided.

New records of Limoniidae and Pediciidae (Diptera) from Croatia

Abstract New records are provided for Limoniidae and Pediciidae from Croatia, with new distribution records for species in 12 different genera. Four genera and 18 species are newly recorded for Croatia. Until now, including this data, 87 Limoniidae and eight Pediciidae have been recorded from Croatia. In this paper we confirm presence of Ormosia (Oreophila) bergrothi (Strobl, 1895) and we give the first records of Dicranota (Paradicranota) pavida (Haliday, 1833) and Molophilus (Molophilus) repentinus Starý, 1971 from Balkan Peninsula.