Ivana Buj

Combining Morphology and Genetics in Resolving Taxonomy - A Systematic Revision of Spined Loaches (Genus Cobitis ; Cypriniformes, Actinopterygii) in the Adriatic Watershed

Taxonomic investigation of spined loaches from Dalmatia and Herzegovina was conducted on specimens from 14 localities. The results of the detailed morphological investigations were combined with genetic data (based on one mitochondrial and two nuclear genes) in order to resolve the taxonomic status of each Cobitis population. Among the investigated features of external morphology, the appearance of spots on the caudal fin base turned out to have the greatest diagnostic value. Furthermore, the number of branched fin rays enabled the discrimination of several species. No morphometric character alone could ensure determination of any Cobitis species. Nevertheless, groups of populations that are more similar in their body shapes correspond to mitochondrial phylogenetic lineages. Based on molecular genetic markers, Dalmatian and Herzegovinian spined loaches form independent lineages inside the Adriatic phylogenetic group. Mitochondrial DNA phylogenetic reconstruction revealed six monophyletic lineages, corresponding to six species distributed in the investigated area. The population distributed in Mostarsko blato karstic field in Bosnia and Herzegovina is described as a new species based on a unique combination of morphological characters: a single triangular Canestrini scale; usually 51/2 branched anal fin rays, 61/2 branched dorsal fin rays, 14 branched caudal fin rays; no spots in the surface pigmentation layer on the caudal fin base; scales on the body very small.

Morphological comparison of bleaks (Alburnus, Cyprinidae) from the Adriatic Basin with the description of a new species

Abstract. The morphometric, meristic and phenotypical characters of the members of the genus Alburnus from the Adriatic Basin were analyzed on specimens from 11 localities, representing eight watersheds. The number of gill rakers, the number of lateral line scales, the number of branched anal fin rays and the coverage of the ventral keel by scales have the greatest significance in differentiating between species. Significant morphological differences exist between the Alburnus population from Lake Lugano (type locality for Alborella maxima Fatio, 1882) and all the remaining investigated populations. Alburnus from the Neretva River drainage is described as a new species and it is distinguished from other species of the genus Alburnus by the following combination of characters: 13½-15½ branched anal fin rays; 17–26 gill rakers; 40–48 lateral line scales; first anal fin ray below branched dorsal rays 6–10 or located up to one scale after the last dorsal fin ray; ventral keel exposed for at least ⅔ of a distance between the pelvic fin base and the anal aperture. The status of the Neretva River basin populations as a separate species was corroborated by genetic analyses, with p-distance between newly described species and A. arborella from the Zrmanja River 1.8–2.2%.

Different Histories, Different Destinies‒Impact of Evolutionary History and Population Genetic Structure on Extinction Risk of the Adriatic Spined Loaches (Genus Cobitis; Cypriniformes, Actinopterygii).

The region of Balkans is often considered as an ichthyologic “hot spot”, with a great number of species and high portion of endemics living in fresh waters in a relatively small area. The Adriatic watershed in Croatia and Herzegovina is inhabited by six spined loach species (genus Cobitis) whose extinction risk estimations were based solely on their extent of occurrence (and/or area of occupancy) and its fragmentation, and conservation proposals do not consider diversity below species level. In this investigation we employed molecular genetic methods to describe present genetic structure of the Adriatic spined loaches and reveal their demographic history. The divergence of the Adriatic lineages inside the genus Cobitis started in Miocene and lasted until Pleistocene epoch. Geological events responsible for shaping recent diversity of spined loaches in the Adriatic basin are: the Dinarid Mountains upwelling, the evolution of Dinaric Lake system, local tectonic activity, river connections during glaciations and differences in sea level. Even though all the investigated species inhabit karstic rivers located in the same geographic area and that were subject of similar geological events, the results obtained reveal great differences in their genetic diversity and structure and point out the necessity of different conservation measures to ensure their future viability. High level of genetic polymorphism is characteristic for species located more to the south. Two species comprised of more than one population have completely different intraspecific structure; populations of C. illyrica are genetically distinct and represent separate evolutionary significant units, whereas intraspecific structure of C. narentana corresponds to metapopulational pattern. Without population genetic data, evolutionary significant units could be easily misidentified. Furthermore, the obtained results affirm that population genetic measurements are able to detect differences among closely located and related species and estimate extinction risk even more accurately than currently applied IUCN criteria.

A new endemic cyprinid species from the Danube drainage.

Populations of endemic Croatian dace were found to belong to two different species, one of which is first described in this study. Telestes karsticus sp. nov. differed morphologically from Telestes polylepis in the total count of lateral line scales, number of gill rakers and the shape of the posterior margin of the anal fin. Morphological differences were corroborated with mtDNA analyses (with p-distance between T. polylepis and T. karsticus sp. nov. ranging between 3·2 and 4·1%; and the number of substitutions between 37 and 47). The newly described species is geographically very localized. It has been recorded from only four localities around Velika Kapela and Mala Kapela mountains in Croatia.

Sexual dimorphism of five Cobitis species (Cypriniformes, Actinopterygii) in the Adriatic watershed

Abstract. Morphological differences between female and male spined loaches belonging to five species from the Adriatic basin were examined. Besides the presence of the Canestrini scale and the length of the pectoral fins, sexual dimorphism is also visible in the position of the pectoral fins, as well as in the length of the pelvic fins. Both pectoral and pelvic fins are significantly longer in males than in females, whereas pectoral fins of females are located more anteriorly then in males. Differences in the length of pectoral and pelvic fins among species are also revealed. The role of the secondary sexual characters in males is probably connected with their special mating behaviour. Furthermore, a hypothesis that habitat conditions have greater impact on body dimensions of females than males is proposed.

FIRST RECORD OF THREE-SPINED STICKLEBACK (Gasterosteus aculeatus Linnaeus, 1758) IN THE DANUBE BASIN OF CROATIA

The three-spined stickleback (Gasterosteus aculeatus Linaeus, 1758) is a small teleost fish considered to be one of the most widely distributed freshwater fishes in the world (Morrow, 2008). In the northern hemisphere, sticklebacks are absent only along the Arctic coasts of Siberia and North America (Mrakovcic et al., 2006). In Europe they often form highly abundant populations, and in some ecosystems are dominant (Clavero et al., 2009). Three-spined sticklebacks are listed as a species of Least Concern (LC) in the European IUCN Red List (Freyhof et al., 2008). However, freshwater populations in the Mediterranean region have experienced a sharp decline or even local extinction events (Clavero et al., 2009). Despite their wide global distribution, stickleback populations in Croatia are present in a fragmented distribution in isolated rivers of the Adriatic Basin: the Krka River (Visovac Lake), Neretva River catchment and Mirna River catchment on the Istrian peninsula (Mrakovcic et al., 2006; Zanella, 2009; Zanella et FIRST RECORD OF THREE-SPINED STICKLEBACK (Gasterosteus aculeatus Linnaeus, 1758) IN THE DANUBE BASIN OF CROATIA

Threatened fishes of the world: Aulopyge huegelii Heckel, 1842 (Cyprinidae)

Common names: Oštrulja (Cro); Dalmatian Barbelgudgeon (Eng). Conservation status: IUCN Red List: endangered (Crivelli 2005); Croatia: endangered (Mrakovčić et al. 2006). Identification: D III/(6)7-8 (9), V II/5–8, A II/4–6(7), P I/(13)14–15(16), C 19; pharyngeal teeth 4-4. Size to 25 cm, usually to 20 cm. Head extended, pointed and narrow. Two pairs of barbels on the lips. Coloration silvery to yellow-grey, with brown-black specks on dorsal and lateral areas. Generally without scales, some specimens display individual scales. Lateral line wavy. Final dorsal fin ray serrated. The urogenital papilla in females elongated and connected to the anal fin (Heckel and Kner 1858; Guzina 2000). Distribution: Endemic to Dinaric karst rivers and lakes of Croatia and Bosnia and Herzegovina (Vuković and Ivanović 1971; Mrakovčić and Mišetić 1990). Abundance: Previously quite abundant, though the population has been reduced in recent years (Mrakovčić et al. 2006). Habitat and ecology: Prefers lake habitats. Lives in schools that migrate to subterranean waters in the autumn or during droughts (Vuković and Ivanović 1971). Omnivorous (algae, zooplankton and benthic invertebrates). Lives up to 8 years (Kaćanski et al. 1978; Guzina 2000). Reproduction: Spawning begins in April in shallow waters at temperatures of 13–17° C. Fecundity ranges from 1,700 to 17,000 eggs, depending on size and age of female (Aganović 1970; Guzina 2000). Females lay eggs with ovipositor in crevices between rocks and plant roots. Larvae are benthic and solitary and hatch at 6.5 mm (Freyhof 1997; Bless and Riehl 2002). Threats: Aulopyge huegelii is threatened due to its restricted and isolated distribution, dam construction, habitat destruction, water pollution and introduction of alien species (Crivelli 2005; Mrakovčić et al. 2006). Conservation action: Strictly protected in Croatia. Not protected in Bosnia and Herzegovina. Conservation recommendations: Urgent habitat protection with bans on further regulation of the watercourse and water pollution. Population monitoring recommended. Environ Biol Fish (2009) 85:21–22 DOI 10.1007/s10641-009-9445-z

Status and distribution of spined loaches (Cobitidae) and stone loaches (Nemacheilidae) in Bosnia and Herzegovina

Abstract. This paper presents a critical assessment of the current status and geographical distribution of loaches (Cobitidae, Nemacheilidae) in Bosnia and Herzegovina. This closes a gap in research knowledge with only general distribution data currently available. In the Danube catchment four cobitid (Cobitis elongata, C. elongatoides, Misgurnus fossilis, Sabanejewia balcanica) and one nemacheilid species (Barbatula barbatula) were recorded, while the Adriatic Sea catchment hosts three spined loaches: C. narentana, C. illyrica, and C. herzegoviniensis and nemachilid B. barbatula. Cobitis elongatoides was found to be the most common and distributed throughout the northern Bosnia and Herzegovina; i.e. in the rivers Sava, Drina, Bosna, Vrbas, Una (Danube catchment). Sabanejewia balcanica was recorded in the same area but less frequently. Recent bibliographic evidence forces us to exclude C. taenia from the current checklist. On the other hand, cobitid species from the Adriatic Sea catchment are endemic with a very small distribution. Also, the extinction risk status of each taxon is mentioned following IUCN categories. This study provides geographically constrained information on the recent status of loach populations in the freshwaters of Bosnia and Herzegovina. Further data on the status and geographical distribution would improve the conservation and management of these fishes.

Threatened fishes of the world : Orsinogobius croaticus (Mrakovčić, Kerovec, Mišetić and Schneider, 1996) (Teleostei: Gobiidae)

Abstract Orsinogobius croaticus is a freshwater fish with a small and limited distribution range in the Adriatic basin in Croatia and Bosnia-Herzegovina, where it inhabits cold karstic rivers and springs. The main threats to its populations include habitat fragmentation, declining habitat quality and the introduction of alien species. Conservation recommendations are proposed to improve effective habitat preservation and to expand the knowledge about this species.

THREATENED FISHES OF THE WORLD: Delminichthys krbavensis (Zupani and Bogutskaya, 2002) (Cyprinidae)

Krbava minnow, Delminichthys krbavensis, is freshwater fish species of endemic to the Adriatic watershed in Croatia. It lives only in karstic springs, streams and subterranean waters in Krbavsko polje. This species is protected by Croatian law and it is listed on IUCN Red List as Critically Endangered (CR). D. krbavensis has very limited distribution what makes it vulnerable to different kinds of threats such as water extraction, exploitation of karstic springs and streams, agricultural pollution, road construction and introduction of nonindigenous fish species. Suggested conservation actions for this species are detailed study of biology, ecology and distribution as well as bans on watercourse regulation and road construction, reduction of pollution and restriction of introductions of nonindigenous fish species. Also, conservation of this species should be ensured through implementation of management and action plans.