Growth patterns of the golden loach, Sabanejewia balcanica were examined in the Rijeka River in Central Croatia on a total of 77 specimens collected in a single sampling session in June 2006. No significant difference was found in total body length between males and females. Age was determined from otoliths. Both males and females live 4 years, though due to the sampling period, no 0+ age fish were found in this study. The length-weight relationship was calculated as W=2x10-6TL3.3229 for males and W=3x10-6TL3.2811 for females, indicating positively allometric growth. According to the growth factor k, growth of males was three times faster than that of females.
An assessment of DNA integrity in erythrocytes using the alkaline comet assay was carried out to estimate the impacts of water pollution on Balkan loaches (Cobitis elongata Heckel et Kner, 1858) inhabiting the Sava and Kupa Rivers, rivers of varying different water quality. The amount of DNA damage in cells was estimated from three different parameters comet tail length as the extent of genetic material migration, tail intensity (% DNA in comet tail) and tail moment. The results suggest a genotoxicity of the aquatic environment in the Sava River and demonstrated significantly lower levels of DNA damage in fish captured from the Kupa River. This study confirmed that the comet assay, applied to fish erythrocytes, is a useful tool in determining potential genotoxicity of water pollutants. Although a good DNA damage pattern for Balkan loach was obtained, due to its global and regional conservation status, only restricted use of a small number of specimens per sampling site could be permitted.
Nine species of the family Cobitidae live in Croatia. Seven species belong to the genus Cobitis and one each to the genera Sabanejewia and Misgurnus. Species such as the C. jadovaensis Mustafić et Mrakovčić, 2008, C. bilineata Canestrini, 1866, C. dalmatina Karaman, 1928, C. illyrica Freyhof et Stelbrink, 2007 and C. narentana Karaman, 1928 live in the Adriatic River Basin (the northern Mediterranean), while the C. elongatoides Bacescu et Maier, 1969, C. elongata Heckel et Kner, 1858, S. balcanica Karaman, 1922 and M. fossilis Linnaeus, 1758 inhabit watercourses of the Danube River Basin. Although most Adriatic taxa appear to be endemic to this area, some researchers have revealed new phylogenetic relationships, suggesting the possibility of more recent taxa exchange with Danube freshwater fish fauna. All members of these genera inhabit either running and/or standing waters. According to IUCN methodology, S. balcanica, C. elongata and M. fossilis in the Danube watershed and C. dalmatina and C. narentana in the Adriatic watershed have been classified as vulnerable. C. elongatoides is in the lower risk category. The newly determined species C. bilineata, C. illyrica and C. jadovaensis will be in the endangered category. Here we present the majority of a total of 20 areas and water bodies included in the proposal of Natura 2000 and which have been selected for the conservation of the species from the family of the Cobitidae.
Using mitochondrial DNA (mtDNA) variations, the phylogenetic position of Croatian populations of spined loaches within the European genus Cobitis was assessed. Spined loaches from the Matica, Neretva, Cetina, Zrmanja, Jadova, Sava, Drava and Kupa Rivers are included in two previously described groups: a) Cobitis sensu stricto group, and b) Adriatic group. The Danubian populations of C. elongatoides and its hybrids from the Kupa and Drava Rivers are related to the species included in the Cobitis s. str. group, whereas all other analysed populations clustered within the Adriatic group, which is divided into the “Bilineata”, “Elongata” and “Ohridana-zanandreai” clades. The Croatian spined loaches from the Adriatic watershed are included in the first two clades. Four mitochondrial lineages were revealed within the “Bilineata” clade: 1. ”C. bilineata” lineage containing loaches from the Zrmanja River and Italian and Spanish representatives of C. bilineata; 2. “Neretva-Cetina” lineage included loaches from the Cetina River (C. dalmatina) and the Neretva River (C. narentana); 3. “Jadova” lineage included specimens from the Jadova River; and 4. “Matica” lineage comprised spined loaches from the Matica River. Cobitis elongata from the Kupa and Drava Rivers belonged to the “Elongata” clade with two sublineages. Allozyme analyses of the C. taenia complex revealed the presence of both C. elongatoides and its all-female triploid hybrids in the Danube basin of Croatia.
Taxonomic analyses of four Sabanejewia balcanica populations from Croatia included morphological (morphometric, meristic, phenotypical) and mitochondrial DNA analyses. Comparative analyses of 24 morphometric characters revealed some differences between populations. The specimens from the Drava River are significantly larger than specimens from other populations, whereas the individuals from the Voćinska River represent the smallest European S. balcanica specimens. Due to similarities in morphometric ratios, in the Tree Diagram of Euclidean distances, the Drava and Petrinjčica Rivers populations formed a separate cluster, while the populations from the Rijeka and Voćinska Rivers formed a second cluster. These morphometrical differences are also corroborated by t-test results. Differences among fishes from different populations also exist in external morphology characters, such as spots on the caudal peduncle and the position of the suborbital spine. Mitochondrial DNA analyses enabled us to infer the phylogenetic placement of four Croatian populations within the genus Sabanejewia. All Croatian samples clustered within the two sublineages of the Danubian-Balkanian complex. Samples from the Voćinska River, as well as one sample from the Drava River were included in the “S. balcanica” (VI) sublineage. The remaining samples, including the two remaining haplotypes from the Drava River, were comprised within “S.montana-S.bulgarica-S.balcanica” (III) sublineage.
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 fi n rays and the coverage of the ventral keel by scales have the greatest signifi cance in differentiating between species. Signifi cant 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: 131/2-151/2 branched anal fi n rays; 17-26 gill rakers; 40-48 lateral line scales; fi rst anal fi n ray below branched dorsal rays 6-10 or located up to one scale after the last dorsal fi n ray; ventral keel exposed for at least 2/3 of a distance between the pelvic fi n 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%.
A new spined loach species Cobitis jadovaensis is described from the Jadova River in Croatia. This species differs from its congeners in the Croatian Adriatic basin with a unique set of characters: a very short dorsal fin base; short anal fin base; narrow head; a single lamina circularis; a single small prominent dark inclined spot on the upper part of the caudal base; all four Gambetta zones well developed and reaching beyond the dorsal base; zones Z1 and Z3 with many irregular spots; zone Z1 narrower than Z2; zone Z2 as wide as Z3; zone Z2 with spots larger than a pupil size fuses with Z3 on caudal peduncle; zone Z4 wider than Z2 and Z3, with 11 – 14 roundish to oval blotches.
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.