Abstract. Chromosome number, karyotype formula, C-banding pattern, genome size and DNA base composition were studied in three species of Hyalidae and seven species of Talitridae. A karyotype of 25 chromosome pairs, with median centromeres (FN = 100), was found in all the species of Talitridae analysed and Apohyale prevostii. Genome size (C-value) varies among Talitrida from 0.94 pg in Apohyale crassipes to 2.81 pg in Orchestia gammarellus, and the percentage of AT-DNA in the whole genome ranges from 56.12% in A. crassipes to 68.17% in Sardorchestia pelecaniformis. In comparison with Hyalidae, Talitridae show more uniformity in chromosome number and karyotype formula, and have larger genomes. There is a direct correlation between total DNA content and the amount of C-heterochromatic DNA. The cytogenetical data on Talitrida were compared from a phylogenetic and an evolutional point of view. The increase in genome size during the evolution of the Talitrida possibly had a role in their adaptation to supralittoral life and extreme subaerial conditions.

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A cytogenetical study was carried out on 34 specimens of the European bitterling Rhodeus amarus (Teleostei: Cyprinidae, Acheilognathinae) from four rivers of the Venice district (NE Italy). This allochthonous fish species was accidentally introduced in the North-East of Italy about 20 years ago and is now rapidly spreading all over the rivers of the Northern part of the country. All the studied specimens are characterised by the same karyotype (2n = 48: 8M + 20SM + 20ST), i.e., the typical one of the native populations of the species. However, a polymorphism in the number of NOR bearing chromosomes has been found. In fact, in addition to the main species-specific NORs, on the short arms of chromosome pair 7, two to five additional 18S rDNA sites have been revealed by FISH in different specimens. Sequential staining with silver nitrate, chromomycin A 3 and DAPI revealed that most of the additional sites are inactive and CMA 3 -positive.Data herein reported confirm that in spite of an overall morphological karyological conservativeness, significant differences for the finer cytogenetic features can be found within the Acheilognathinae with the 2n = 48 and NF = 76 karyotype.

Genomic In Situ Hybridization (GISH) is a powerful tool to identify and to quantify genomic constituents in allopolyploids, and is mainly based on hybridization of highly and moderate repetitive sequences. In animals, as opposed to plants, GISH has not been widely used in part because there are technical problems in obtaining informative results. Using the allopolyploid Squalius alburnoides Steindachner, 1866 fish complex as a model system, we succeeded in overcoming methodological constraints when dealing with parental species with a small genome size. This hybridogenetic complex has biotypes with different genome compositions and ploidy levels, but parental chromosomes are small, morphologically very similar and therefore cannot be distinguished by conventional cytogenetic approaches. Specimens have a small genome (C-value1.2 pg) with a low level of highly and moderate repetitive sequences, mainly located at pericentromeric chromosome regions. Since it is well known that probe annealing depends on probe concentration and hybridization time to obtain uniform hybridization signals along the chromosome arms, we progressively increased the amount of labeled probes from 100ng up to 1µg and the incubation time from overnight up to 5 days. We also made other smaller improvements. Results showed a clear enhancement of signals with respect to previous data, allowing an accurate and reproducible assignment of the parental genomes in both diploid and triploid fish.It was thus evidenced that high probes’ concentrations and long incubation time are the key to obtain, without extra image editing, uniform and reliable hybridization signals in metaphase chromosomes of animal hybrids from species with small genome size.

In this study, the description of the karyotypes of the endangered chubs Squalius aradensis (Coelho, Bogutskaya, Rodrigues and Collares-Pereira, 1998) and Squalius torgalensis (Coelho, Bogutskaya, Rodrigues and Collares-Pereira, 1998) is presented by means of conventional (Giemsa-staining, Chromomycin A3 (CMA3)-fluorescence, Silver-impregnation (Ag-NORs)) and molecular (fluorescence in situ hybridization (FISH) with 18S rDNA probe) protocols. These endemic sister-species have an allopatric but adjacent distribution in the most southwestern part of the Iberian Peninsula. Diploid chromosome number was invariably 2n = 50 and karyotypes of both species were grossly similar, composed of metacentric and submetacentric elements with a reduced number of acrocentric pairs. Sequential staining using FISH with an 18S rDNA probe, CMA3 and Ag-NORs treatments revealed consistent positive signals located at the end of the short arms of a submetacentric chromosome pair, likely homologous in both species. While providing useful cytogenetic comparative data against other members of the genus Squalius Bonaparte, 1837, the work aimed to draw attention towards the conservation of two narrow-range and highly confined fish species.