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Polyploidy is exceptional in mammals and rare in reptiles. Unscheduled polyploidy often leads to cell disorders and cancer through genomic rearrangements that can be difficult to trace and is particularly poorly studied in non-model vertebrates. In the present paper, using a combination of banding and Fluorescence In Situ Hybridization (FISH) techniques, we reconstruct and characterize the origin and significance of aneupolyploid karyotypes found in the bone marrow of an individual of the Malagasy gecko Geckolepis typica. Our results demonstrate that these anomalous karyotypes correspond to a proliferative cell disorder characterized by a succession of distinct structural events, from a preliminary duplication of the whole genome to the instability of loci of NORs and subsequent chromosome rearrangements that will produce aneuploid cells. Our comparison between the newly described normal, i.e., diploid, karyotype of the species (2n = 40) and aneutetraploid plates (4n = 77–80) revealed that translocations of NOR-bearing chromosomes are involved in generating aneuploidy. Furthermore, simultaneous and multiple translocations of the NOR-bearing chromosomes occurred with relatively high frequency (27% and 19%, respectively) on distinct chromosome pairs. In addition, in about 9% of 90 scored metaphase plates, structural instability of the NOR regions also produced detachments of rDNA arrays and generated an accessory, fully heterochromatic B-chromosome. Our data suggest that the duplication of the whole genome can be considered a preliminary stage in proliferative cell disorders, inducing chromosomal instability and structural rearrangements. © 2018 Deutsche Gesellschaft für Herpetologie und Terrarienkunde e.V. (DGHT), Mannheim, Germany.

Sexual differentiation (SD) during development results in anatomical, metabolic, and physiological differences that involve not only the gonads, but also a variety of other biological structures, such as the brain, determining differences in morphology, behavior, and response in the breeding season. In many reptiles, whose sex is determined by egg incubation temperature, such as the leopard gecko, Eublepharis macularius, embryos incubated at different temperatures clearly differ in the volume of brain nuclei that modulate behavior. Based on the premise that "the developmental decision of gender does not flow through a single gene", we performed an analysis on E. macularius using three approaches to gain insights into the genes that may be involved in brain SD during the thermosensitive period. Using quantitative RT-PCR, we studied the expression of genes known to be involved in gonadal SD such as WNT4, SOX9, DMRT1, Erα, Erβ, GnRH, P450 aromatase, PRL, and PRL-R. Then, further genes putatively involved in sex dimorphic brain differentiation were sought by differential display (DDRT-PCR) and PCR array. Our findings indicate that embryo exposure to different sex determining temperatures induces differential expression of several genes that are involved not only in gonadal differentiation (PRL-R, Wnt4, Erα, Erβ, p450 aromatase, and DMRT1), but also in neural differentiation (TN-R, Adora2A, and ASCL1) and metabolic pathways (GP1, RPS15, and NADH12). These data suggest that the brains of SDT reptiles might be dimorphic at birth, thus behavioral experiences in postnatal development would act on a structure already committed to male or female.

The faunistic data on the recent land molluscs of Vesuvius National Park (Naples, Southern Italy) and neighbouring areas sampled during field surveys from 2005 to 2006 are reported here. The data were obtained through a careful research on museum collections and literature and were supplemented with observations from field surveys. Up to now, only 23 land snails species were known from the Vesuvius National Park through bibliographical data. The faunistic research, carried out in 25 stations of different environment typologies, allowed us to discriminate 31 species: 15 of these species are reported here for the first time for the study area. The presence of 16 species was confirmed by other field investigations. Other 19 taxa are reported in the literature, but not confirmed in our research - Helix turriculatella O. G. Costa, 1839 (obsolete nomenclature), Trochoidea trochoides Poiret, 1789, Hygromia cinctella (Draparnaud, 1801), Campylaea planospira (Lamarck, 1822), Vitrea sp. (cf. contracta (Westerlund, 1871)), Cecilioides acicula (O. F. Müller, 1774), Cochlicella acuta (O. F. Müller, 1774), Theba pisana O. F. Müller, 1774). Another 12 taxa are highly doubtful, because of possibilities for confusion with species living in other European or Italian regions, as well as previous misidentifications (Deroceras agreste (Linnaeus, 1758), Monacha gregaria (Rossmässler, 1839), Cochlicella barbara (Linnaeus, 1758), Monachoides incarnatus (O. F. Müller, 1774),Cernuella (Cernuella) cisalpina (Rossmässler, 1837), Cernuella aradasii (Pirajno, 1842), Oxychilus cellarius (O. F. Müller, 1774), O. alliarius (J. S. Miller, 1822), Zonitoides nitidus (O.F. Müller, 1774), Hyalina pseudohydatina, Ferussacia lubrica, Aegopinella nitens (Michaud, 1831); it was impossible in these cases to refer to their current specific name. Eobania vermiculata (O. F. Müller, 1774), Rumina decollata (Linnaeus, 1758), Papillifera papillaris (O. F. Müller, 1774) appear to be the most common species. Interesting are the records of Marmorana muralis (O.F. Müller, 1774), Helix cf. ligata (O.F. Müller, 1774) and Charpentiera gibbula gibbula (Rossmässler, 1836). Charpentieria gibbula honii (Tiberi, 1878) needs genetic investigations in order to confirm its taxonomic status. One allochthonous species (Paralaoma servilis (Shuttleworth, 1852)) was surveyed for the first time in the study area. A Red List of Threatened Species is proposed and the species are classified according to the IUCN criteria (Version 2019-2.1). The malacofauna of Vesuvius - Somma Mountain Complex is composed by 38 land mollusc species and represents 36.2% of the total species present in Campania (about 105 species, personal data) and 12.8% of the Italian fauna (about 530 species of land molluscs extrapolated from the list of Bank (2017). It is assumed that the poor biodiversity of the Vesuvian land malacofauna can be attributed essentially to the scarcity of calcareous substrates and the absence of permanent water sources.

We conducted a phylogenetic study through karyological data, by standard staining and Ag-NOR banding, and molecular analysis (by 12S and 16S mitochondrial rRNA genes and nuclear gene C-mos) on 11 species of Malagasy geckos, genus Paroedura, and two relatives (Ebenavia inunguis and Uroplatus phantasticus). Ebenavia inunguis and U. phantasticus had 2n = 36 telocentric elements, NORs on the first chromosome pair in E. inunguis, and on the third chromosome pair in U. phantasticus. All examined Paroedura showed NORs on the smallest chromosome pair; moreover, six of the eleven examined species show a 2n = 36 karyotype, with a pair of metacentrics and 17 telocentric pair. The remaining species exhibited karyotypes with a diploid chromosome number ranging from 2n = 31 to 2n = 38. We assume that these karyotype assemblages derived from the 2n=36 karyotype by cryptic and/or simple rearrangements, such as inversions, fissions and fusions. Furthermore, molecular and/or chromosomal data indicate that Paroedura is a monophyletic genus, in which chromosome rearrangements occurred repeatedly and independently during the specific diversification. Moreover both P. bastardi and P. gracilis in current definitions are paraphyletic assemblages of several related species, since their population proves more closely related to P. ibityensis or P. oviceps than co-specific populations.Key words: chromosome, Gekkonidae, Madagascar, Paroedura, molecular phylogeny.

Sexual differentiation (SD) during development results in anatomical, metabolic, and physiological differences that involve not only the gonads, but also a variety of other biological structures, such as the brain, determining differences in morphology, behavior, and response in the breeding season. In many reptiles, whose sex is determined by egg incubation temperature, such as the leopard gecko, Eublepharis macularius, embryos incubated at different temperatures clearly differ in the volume of brain nuclei that modulate behavior. Based on the premise that "the developmental decision of gender does not flow through a single gene", we performed an analysis on E. macularius using three approaches to gain insights into the genes that may be involved in brain SD during the thermosensitive period. Using quantitative RT-PCR, we studied the expression of genes known to be involved in gonadal SD such as WNT4, SOX9, DMRT1, Erα, Erβ, GnRH, P450 aromatase, PRL, and PRL-R. Then, further genes putatively involved in sex dimorphic brain differentiation were sought by differential display (DDRT-PCR) and PCR array. Our findings indicate that embryo exposure to different sex determining temperatures induces differential expression of several genes that are involved not only in gonadal differentiation (PRL-R, Wnt4, Erα, Erβ, p450 aromatase, and DMRT1), but also in neural differentiation (TN-R, Adora2A, and ASCL1) and metabolic pathways (GP1, RPS15, and NADH12). These data suggest that the brains of SDT reptiles might be dimorphic at birth, thus behavioral experiences in postnatal development would act on a structure already committed to male or female.

A karyological analysis on six Italian populations the slow worm (Anguis veronensis Pollini, 1818) was performed and their genetic differentiation at the mitochondrial 16S rRNA gene fragment from a Spanish sample has been assessed. The Italian populations were karyologically uniform, all showing 2n= 44 elements, of which 20 were macrochromosomes and 24 microchromosomes. Comparison with literature data on Central European populations showed a difference on the morphology of the 10th chromosome pair: submetacentric in Italian populations and telocentric in the Central European ones. Our analysis showed the presence of a fragile site on chromosomes of this pair, suggesting its propensity for structural rearrangements. Analysis of the 16S rRNA gene fragment showed uniformity among Italian populations (uncorrected genetic distance of 0.4%), and their genetic distinctness from the Spanish individual (uncorrected genetic distance of 4.2%). Our results confirm the existence of two different Anguis fragilis Linnaeus, 1758 lineages, each one characterized by a different cytotype.

A cytogenetic study was carried out by means of standard and NOR-FISH staining methods on the chromosomes of four Alopiinae door snails, two species of the Delimini tribe (Siciliaria paestana and Papillifera bidens) and two species of Medorini tribe (Leucostigma candidescens and Medora sp.). The two Delimini species and Medora sp. showed similar karyotypes, concerning the number of chromosomes, 2n = 62, and the first metacentric pair of chromosomes clearly bigger than the other pairs, but differed in the number of the telocentric pairs: none in P. bidens, four in S. paestana and, even, 21 in Medora sp. Leucostigma candidescens had 2n = 48, all biarmed chromosomes. Loci of NORs were on a single medium small chromosome pair in all the four studied species. The obtained results evidence a relevant degree of intergeneric karyotypic diversity in Alopiinae, concerning both chromosome number and/or shape. The data allow us to make a hypothesis on the evolutionary chromosome trends for this door snail subfamily.

Blindsnakes are poorly studied reptiles from the karyological point of view since chromosomal data are only known for eight out of more than 420 species. The present paper shows the results of a karyological study conducted by means of standard and banding staining (Ag-NOR-banding and C-banding plus CMA and DAPI fluorochromes) on three typhlopid species, Madatyphlops arenarius, Xerotyphlops vermicularis and the parthenogenetic blindsnake, Indotyphlops braminus . Madatyphlops arenarius has conserved the primitive snake karyotype of 2n = 36 with 16 biarmed macro- and 20 microchromosomes and NORs on a microchromosome pair. The karyotype of X . vermicularis differed in showing a reduced number of microchromosomes (16) and NORs on the long arms of the third macrochromosome pair. The parthenogenetic I . braminus exhibited a triploid karyotype of 3n = 42 with seven macro- and seven microtriplets and NORs on a microtriplet. Comparison with chromosome data from the available literature suggests that the primitive snake karyotype of 2n = 36 occurred in the common ancestor of Leptotyphlopidae + Typhlopidae and in the common ancestor of Typhlopidae. In the latter family chromosome evolution may well have proceeded towards a decrease in the chromosome number from 2n = 36, 34, 32 until it reached 2n = 28, mainly involving micro- to microchromosome and/or macrochromosome translocations. Furthermore, these chromosome rearrangements occurred independently and repeatedly in the different evolutionary lineages of this family and are partially concordant with phylogenetic relationships. Cytologically identifiable sex chromosomes were not revealed by either standard or C-banding staining.

The stranding of a suckling calf of Common Minke Whale (Balaenoptera acutorostrata) on the coast near Salerno (Campania, Southern Italy) is reported. The molecular analysis of a partial sequence of the mitochondrial DNA control region shows that the animal bore a haplotype identical to haplotype Ba169 considered as typical of individuals from North Atlantic population. Historical data and our results suggest the possibility that the Mediterranean Sea might be a potential calving or nursery ground for this species.

Within the scope of a project on the characterization of satellite DNAs in polar mollusks, the Antarctic chiton Nuttallochitonmirandus (Thiele, 1906) was analyzed. Two novel families of tandemly repeated DNAs, namely NmH and NmP, are described in their structure and chromosomal localization, and, furthermore, their presence was analyzed in related species. Data reported here display a particular variability in the structural organization of DNA satellites within this species. Processes driving satellite evolution, which are likely responsible for the intriguing variability of the identified satellite DNAs, are discussed.

A repeated DNA sequence, named RepHv-390, isolated from DNA samples of male and female specimens of the European Western wipe snake, Hierophis viridiflavus, was used as probe for FISH analysis to detect its distribution on chromosomes of this colubrid species and of other species of the same and of different snake families. We observed positive results in all the studied species, namely: H. viridiflavus, Coronella austriaca, Elaphe quatuorlineata (Colubridae), Phisalixella variabilis (Lamprophiidae) and Vipera aspis (Viperidae). Intense and abundant hybridization signals of RepHv-390 were observed on all chromosomes of H. viridiflavus, C. austriaca, E. quatuorlineata and V. aspis and on nuclei of P. variabilis. Its wide presence on the chromosomes of all the studied species suggests that RepHv-390 is a segment of transposable element (TE) of ancient origin in snakes. Probably, the ancient TE arose in a common Colubroidea ancestor before the diversification into different lineages of this superfamily. Preliminary analysis of RepHv-390 evidenced no significant identity with any other known sequence, suggesting that RepHv-390 is a segment of an unclassified TE element

Karyological studies continue to clarify the evolutionary history of many taxa. We report here on the first chromosome survey of the frog Pseudhymenochirus merlini (Pipidae) in comparison with the related species Hymenochirus boettgeri . In addition to the 2 n = 20 regular (A) chromosomes (eight metacentric and two subtelocentric pairs), H. boettgeri has an accessory B chromosome in about half of the scored karyotypes (2 n = 21). P. merlini showed a karyotype of 2 n = 20 chromosomes, differing from the set of A chromosomes of H . boettgeri by three inversions. Other differences are in the location of the nucleolus organizer regions (NORs), on the 10th chromosome pair in P. merlini instead of the 4th pair in H. boettgeri . Heterochromatin is less abundant in P. merlini , and localized in centromeric and telomeric regions of all chromosomes. The patterns in H. boettgeri resemble those of another pipid species, Silurana tropicalis , as to chromosome number (2 n = 20), morphology, and G-banding patterns. We propose that these characteristics are symplesiomorphic in Pipidae, allowing us to present a scenario for the chromosome evolution in this family. Our results identify a third mechanism of increase in chromosome numbers among pipid frog species, the addition of B chromosomes in Hymenochirus , besides the known processes of polyploidization in Xenopus / Silurana , and centric fission in Pipa .

The aim of this work is to investigate the sequence conservation and the evolution of repeated DNA in related species. Satellite DNA is a component of eukaryotic genomes and is made up of tandemly repeated sequences. These sequences are affected by high rates of mutation that lead to the occurrence of species-specific satellite DNAs, which are different in terms of both quantity and quality. In this work, a novel repetitive DNA family, named PjHhaI sat, is described in Pecten jacobaeus. The quantitative analyses revealed a different abundance of this element in the molluscan species investigated in agreement with the "library hypothesis" even if, in this case, at a high taxonomic level. In addition, the qualitative analysis demonstrated an astonishing sequence conservation not only among scallops but also in six other molluscan species belonging to three classes. These findings suggest that the PjHhaI sat may be considered as the most ancients of DNA described so far, which remained "frozen" during molluscan evolution. The widespread distribution of this sat DNA in molluscs as well as its long evolutionary preservation open up questions on the functional role of this element. A future challenge might be the identification of proteins or molecules which interact with the PjHhaI sat.

Glycoconjugates secreted by the pedal system of the rayed limpet, Patella caerulea, were characterised in situ by histochemical and lectin-histochemical methods in individuals collected around the annual cycle, in November, March, and June. Stainings with periodic acid-Schiff (PAS), Alcian blue pH 2.5 (AB pH 2.5), Alcian blue pH 1.0 (AB pH 1.0), high-iron diamine-Alcian blue pH 2.5 and lectin binding assays with 9 lectins (Con A, WGA, succinylated-WGA, PNA, DBA, SBA, AAA, UEA-I, LTA) were performed. Four secreting cell types were observed in the sole, one in the peripheric region, and two in the sidewall. Glycoconjugate composition varied among cell types and also in one and the same cell type throughout the year. β-Elimination followed by PAS and AB pH 2.5 stainings indicated that most saccharidic chains were O-linked to the protein backbone. Secretion by sole and peripheric region was acidic, carboxylated and/or sulfated, whereas that of the sidewall was neutral. Glucosaminylated and 1,4-fucosylated residuals were predominant in the cell types along the year, 1,2-fucosylated residuals being observed only in the sidewall cells in June. Mannosylated and/or glycosylated residuals were observed in all cells mostly in November. Galactosylated/galactosaminylated residuals were present mostly in the sidewall cells and in the sole subepidermal mucocytes in June. Mannosylated and/or glycosylated residuals in November are probably linked to gonad maturation or to higher locomotion and foraging activity, whereas galactosaminylation in the sole cells and 1,2-fucosylation and glucosaminylation in the sidewall cells in June are linked to a prolonged stationary state, increasing water adsorption to counteract dehydration and/or to modulate microbial interactions.

The stranding of a suckling calf of Common Minke Whale (Balaenoptera acutorostrata) on the coast near Salerno (Campania, Southern Italy) is reported. The molecular analysis of a partial sequence of the mitochondrial DNA control region shows that the animal bore a haplotype identical to haplotype Ba169 considered as typical of individuals from North Atlantic population. Historical data and our results suggest the possibility that the Mediterranean Sea might be a potential calving or nursery ground for this species.

Sexual differentiation (SD) during development results in anatomical, metabolic, and physiological differences that involve not only the gonads, but also a variety of other biological structures, such as the brain, determining differences in morphology, behavior, and response in the breeding season. In many reptiles, whose sex is determined by egg incubation temperature, such as the leopard gecko, Eublepharis macularius, embryos incubated at different temperatures clearly differ in the volume of brain nuclei that modulate behavior. Based on the premise that "the developmental decision of gender does not flow through a single gene", we performed an analysis on E. macularius using three approaches to gain insights into the genes that may be involved in brain SD during the thermosensitive period. Using quantitative RT-PCR, we studied the expression of genes known to be involved in gonadal SD such as WNT4, SOX9, DMRT1, Erα, Erβ, GnRH, P450 aromatase, PRL, and PRL-R. Then, further genes putatively involved in sex dimorphic brain differentiation were sought by differential display (DDRT-PCR) and PCR array. Our findings indicate that embryo exposure to different sex determining temperatures induces differential expression of several genes that are involved not only in gonadal differentiation (PRL-R, Wnt4, Erα, Erβ, p450 aromatase, and DMRT1), but also in neural differentiation (TN-R, Adora2A, and ASCL1) and metabolic pathways (GP1, RPS15, and NADH12). These data suggest that the brains of SDT reptiles might be dimorphic at birth, thus behavioral experiences in postnatal development would act on a structure already committed to male or female.

The two endemic Italian brown frog species, Rana latastei and R. italica, have a very similar morphology: the diagnostic characters are the tympanum, larger in R. latastei and the nostril distance that is almost equal to the interorbital length in R. latastei, while is slightly greater in R. italica. The most important parameter in order to discriminate between the two species is their different areals: R. latastei is mostly widespread in the Po Valley, while R. italica inhabits almost only the Apennine rivers. The S1 satellite DNA from Palaearctic brown frogs displays a different structure in all the previously tested species, and this complete species- specificity was verified also for R. latastei and R. italica. Southern Blot of DNA samples from the two species, digested with five different restriction enzymes, displays different patterns of hybrid bands. Both species present two repetitive units S1a and S1b. The S1a repetitive unit has the same size but different restriction sites, while the S1b unit differs both in size and restriction sites. In conclusion, the S1 satellite DNA is a powerful molecular marker in order to discriminate the Palaearctic brown frog species.

Within the scope of a project on the characterization of satellite DNAs in polar mollusks, the Antarctic chiton Nuttallochitonmirandus (Thiele, 1906) was analyzed. Two novel families of tandemly repeated DNAs, namely NmH and NmP, are described in their structure and chromosomal localization, and, furthermore, their presence was analyzed in related species. Data reported here display a particular variability in the structural organization of DNA satellites within this species. Processes driving satellite evolution, which are likely responsible for the intriguing variability of the identified satellite DNAs, are discussed.

A karyological study conducted by both conventional and banding staining methods (Ag-NOR-, CMA3, and C-banding) evidenced a peculiar heterochromatin pattern on chromosomes of Moldovan specimens of the agile frog, Rana dalmatina. As is normal for all agile frogs, the Moldovan population presented 2n = 26 chromosomes, with NOR loci on the short arms of the third chromosome pair, but differed in showing heavy centromeric, CMA3 positive C-bands on the seventh chromosome pair, and light centromeric, DAPI positive C-bands on five chromosome pairs. In contrast, Moldovan specimens showed no significant difference in two segments of the 16S mitochondrial rRNA gene and in the S1 satellite DNA sequence and organization, in comparison with the GenBank deposited 16S rDNA and S1 DNA satellite sequences of R. dalmatina. Molecular similarity and chromosomal differences between agile frogs from Moldovan and extra-Moldovan populations are discussed.

Nel presente lavoro abbiamo esaminato la distribuzione e la densità per quadrante degli Anfibi della Campania. Gli Anfibi più comuni nella regione sono Pelophylax sp., Bufo bufo, Rana italica e Lissotriton italicus, rinvenuti in più del 30% dei quadrati UTM 10x10 km investigati. L’Anfibio più raro è Lissotriton vulgaris che è confinato all’area nordoccidentale della regione. Degna di nota è la presenza, anche in sintopia, di Salamandrina perspicillata e Salamandrina terdigitata, al momento unico caso finora noto in Italia.

Anguis fragilis (Squamata: Anguidae). Sono stati studiati vari individui di tre popolazioni italiane di Anguis fragilis mediante metodi di colorazione standard dei cromosomi. Tutti gli individui studiati mostrano un cariotipo di 2n=44 elementi (20 macro- e 24 microcromosomi). Tre coppie di macrocromosomi (1, 4 e 5) sono metacentrici, la decima coppia è formata da cromosomi subtelocentrici, mentre tutti i rimanenti macrocromosomi sono telocentrici. I risultati ottenuti indicano che le popolazioni italiane di orbettino sono cariologicamente uniformi, ma differiscono dalle popolazioni centro-europee, che hanno cariotipi in cui gli elementi della decima coppia sono telocentrici.

To date the knowledge of repeated elements in the snake genome is scarce and mostly limited to various satellite and microsatellite sequences, found in species belonging to different taxonomic groups. In a wider study concerning phylogeography of the European whip snake, Hierophis viridiflavus, we identified three new repeated sequences. Then, we performed a study aimed at characterizing these sequences, underlining their principal features with particular regard to nucleotide composition, genomic quantification and chromosome localization. Two of the three repeated sequences differed in length (240 and 360 bp, respectively), but showed a 5’ segment of about 110 bp with an identity higher than 90%. Moreover this shared segment showed a trait of 85 bp with an identity of 77% with an interspersed L1element of Anolis carolinensis. Both sequences of 240 and 360 bp are organized as interspersed elements and almost exclusively localised on the W sex chromosome, representing about 0,6% of the genome. The third sequence, long about 240 bp, did not show any significant identity with the other two isolated sequences and with any other database deposited sequences. This sequence, too, is an interspersed element, representing about 2% of the genome. Differently from the other two W specific sequences, this third repeated sequence is interspersed on both autosomes and Z W sex chromosomes. We identified three new interspersed repeated sequences in H. viridiflavus. Two of them are W-specific elements related to an L1 elemnt of A. carolenesis. The third sequence is a unknown interspersed element, localised on both autosome and ZW sex chromosomes. Comparative genomic and chromosome hybridizations on other related snakes provided useful systematic and phylogentic information.

Glycoconjugates secreted by the pedal system of the rayed limpet, Patella caerulea, were characterised in situ by histochemical and lectin-histochemical methods in individuals collected around the annual cycle, in November, March, and June. Stainings with periodic acid–Schiff (PAS), Alcian blue pH 2.5 (AB pH 2.5), Alcian blue pH 1.0 (AB pH 1.0), high-iron diamine–Alcian blue pH 2.5 and lectin binding assays with 9 lectins (Con A, WGA, succinylated-WGA, PNA, DBA, SBA, AAA, UEA-I, LTA) were performed. Four secreting cell types were observed in the sole, one in the peripheric region, and two in the sidewall. Glycoconjugate composition varied among cell types and also in one and the same cell type throughout the year. -Elimination followed by PAS and AB pH 2.5 stainings indicated that most saccharidic chains were O-linked to the protein backbone. Secretion by sole and peripheric region was acidic, carboxylated and/or sulfated, whereas that of the sidewall was neutral. Glucosaminylated and 1,4-fucosylated residuals were predominant in the cell types along the year, 1,2-fucosylated residuals being observed only in the sidewall cells in June. Mannosylated and/or glycosylated residuals were observed in all cells mostly in November. Galactosylated/galactosaminylated residuals were present mostly in the sidewall cells and in the sole subepidermal mucocytes in June. Mannosylated and/or glycosylated residuals in November are probably linked to gonad maturation or to higher locomotion and foraging activity, whereas galactosaminylation in the sole cells and 1,2-fucosylation and glucosaminylation in the sidewall cells in June are linked to a prolonged stationary state, increasing water adsorption to counteract dehydration and/or to modulate microbial interactions.

Trochoidea (Trochoidea) caroni (Deshayes, 1832), with T. (T.) elegans (Gmelin, 1791) and T. (T.) trochlea (Pfeiffer, 1846), belong to a group of species (“elegans group”), which, although considered valid by most authors, show a not completely defined systematic position. In this work, we report four new records (Terracina and Spigno Saturnia for Latium, Sorrento for Campania and Castagneto Carducci for Tuscany) and confirm two historical records (Ischia Island for Campania and Manfredonia for Apulia) of T. caroni from Italy, and all published and new faunistic data about the presence of this species in the Sicilian Islands, Capri Island and Maltese Islands. Moreover, we report some historical data from the Western Mediterranean Sea: two bibliographical records from the Balearic Islands, not verified, and three records from North Africa (Tunis in Tunisia, Annaba in Algeria and an unmarked locality in Morocco). We also report the up to now historical faunistic data of T. trochlea, a not well known species with an undefined status, and of some populations of T. elegans with raised whorl. It is not yet possible to draw a certain conclusion on the particular biogeography of these taxa, but the new data presented in this work, enhance the current knowledge on the distribution of T. caroni which would be larger than previously known.

Two new high repetitive DNAs were isolated from Discoglossus pictus genomic DNA digested with StuI. Sequence analysis of clones indicated two types of repetitive units 143–148 bp and 170–177 bp long, respectively. Southern blot analyses show ladders of bands indicating that both repetitive units are organized in long tandem arrays and belong to two different satellite DNAs that were named Dp-sat2 and Dp-sat3. A similar ladder of bands was also produced when genomic DNA was digested with ClaI in the case of Dp-sat2, or with ScaI in the case of Dp-sat3. Quantitative dot-blot analyses indicate that Dp-sat2 and Dp-sat3 account for about 2.6% and 2.2%, respectively, of the D. pictus genome. Considering that D. pictus has a genomic DNA content of 10.5 pg/N, our data indicate that Dp-sat2 and Dp-sat3 repetitive units are present in about 2.3 millions and 1.6 millions copies per diploid genome, respectively. Dp-sat2 and Dp-sat3 probes produced the same patterns of hybrid bands in Southern blots and the same genomic content in dot-blots in all the other Discoglossus species, except D. montalentii which showed a different band pattern and genomic content. The chromosomal distribution of the two StuI satellites shows some similarities: both Dp-sat2 and Dp-sat3 probes mainly labelled the pericentromeric regions of the large chromosome pairs 1–5 and 7. Moreover, probes from both satellite DNAs also produced faint fluorescent mini-spots along almost all chromosome pairs. These data suggest that both satellite DNAs have two types of organization, one in long arrays localized in pericentromeric positions, and the other in short arrays dispersed in the genome.

The present paper shows the results of chromosome banding and rDNA-FISH study performed on several specimens of different populations of Patella caerulea, Patella rustica and Patella ulyssiponensis. The taxonomic attribution of specimens was ascertained by the molecular phylogenetic analysis of the mitochondrial 16S rRNA gene. P. caerulea and P. rustica had 2n = 18 chromosomes with first seven of biarmed pairs and the remaining two uniarmed pairs. P. ulyssiponensis had 2n = 16 with all biarmed chromosomes. Ag-NOR loci were on the short arms of the first metacentric pair in the three studied limpets, whereas they showed a different pattern of heterochromatin distribution and composition. A chromosome mosaicism was observed in several P. caerulea specimens, which exhibited an unpaired metacentric element and loss of a telocentric pair. The obtained results suggest that in the genus Patella specific diversification was accompanied by variations in heterochromatin distribution and composition and reduction of chromosome number by Robertsonian centric fusion.

A karyological study conducted by both conventional and banding staining methods (Ag-NOR-, CMA3, and C-banding) evidenced a peculiar heterochromatin pattern on chromosomes of Moldovan specimens of the agile frog, Rana dalmatina. As is normal for all agile frogs, the Moldovan population presented 2n = 26 chromosomes, with NOR loci on the short arms of the third chromosome pair, but differed in showing heavy centromeric, CMA3 positive C-bands on the seventh chromosome pair, and light centromeric, DAPI positive C-bands on five chromosome pairs. In contrast, Moldovan specimens showed no significant difference in two segments of the 16S mitochondrial rRNA gene and in the S1 satellite DNA sequence and organization, in comparison with the GenBank deposited 16S rDNA and S1 DNA satellite sequences of R. dalmatina. Molecular similarity and chromosomal differences between agile frogs from Moldovan and extra-Moldovan populations are discussed.

Within the scope of a project on the characterization of satellite DNAs in polar mollusks, the Antarctic chiton Nuttallochitonmirandus (Thiele, 1906) was analyzed. Two novel families of tandemly repeated DNAs, namely NmH and NmP, are described in their structure and chromosomal localization, and, furthermore, their presence was analyzed in related species. Data reported here display a particular variability in the structural organization of DNA satellites within this species. Processes driving satellite evolution, which are likely responsible for the intriguing variability of the identified satellite DNAs, are discussed.

By applying advanced techniques that are available now days, it is relatively feasible to determine and monitor the degree of pollution of marine systems. However, it is still difficult to evaluate the effects that pollution may have on marine organisms and, in general, on the environment. A way to overcome this limit is by biomonitoring, i.e. by monitoring the biological responses of selected organisms (namely biomarkers) to pollutants. Many species of molluscs have been labelled as efficient biomarkers. In my research project the aim was to verify the appropriateness of limpets (Patella caerulea) as biomarkers by analyzing specimens of Patella collected in sites with different degrees of pollution along the coasts of the Campania region (Italy). To reach my goal, I measured the potential damage of pollutants at two levels: somatic (by monitoring gill histopathologies) and germinal (by monitoring gonad chromosomic aberrations). My results suggest that pollutants caused a significant damage in natural populations of P. caerulea. The gills of specimens collected from polluted sites presented structural changes, cell and tissue degeneration (cells with vacuoles, dissociation of haemocele septa, nuclear alterations), or the development of compensative mechanisms related to environmental toxicity (epithelium swelling, hyperplasia and fusion of septa). Furthermore, although preliminary, the results suggest that the level of gill injury appeared to be related to the degree of pollution of the study site. Finally, gill aberrations were always higher in each study site compared to the reference site. 2 The gonads of individuals of polluted sites also showed comparatively higher rates of chromosome aberrations, which are supposed to be a by product of an exposition to heavy metal and organic pollutants. In conclusion, the results obtained in my research project suggest that pollutants may act on the cells both of the somatic and germinal lineage in Patella and are thus potentially dangerous for the whole natural population of limpets of the Campania region Dottorato di Ricerca in Biologia Animale,XX Ciclo, a.a.2006-2007 Università della Calabria