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Gli anfibi costituiscono un ottimo modello per esplorare gli aspetti storici della distribuzione delle specie a causa della loro bassa capacità di dispersione e della bassa vagilità individuale. I bufonidi sono una delle famiglie di Anuri più ricche di specie, tra cui il rospo comune, Bufo bufo, diffuso nelle regioni eurasiatiche. Abbiamo eseguito uno studio cariologico con tecniche standard di bandeggio C e sequenziali con fluorocromi (Cromomicina A3 (CMA) e Diamidinofenilindolo (DAPI) su diversi girini di diverse popolazioni del Centro e Sud Italia. Tutti i girini esaminati presentavano il cariotipo Bufo standard, di 2n = 22 cromosomi a due braccia, con le prime sei coppie più grandi delle altre cinque (7 - 11), e l'eterocromatina NOR associata distale sui bracci lunghi della sesta coppia di cromosomi, che era anche l'unica regione cromosomica CMA-positiva. Il bandeggio C ha evidenziato eterocromatina centromerica, DAPI positiva, su tutti i cromosomi in tutte le popolazioni studiate dell'Italia centrale. Le popolazioni dell'Italia meridionale differivano per bande C paracentromeriche aggiuntive sui bracci corti dei cromosomi 1, 3 e 5. Questi risultati supportano la suddivisione delle popolazioni centrali di B. bufo da quelle meridionali, come evidenziato anche da studi filogenetici molecolari., BORNH Bulletin of Regional Natural History, V. 2 N. 3 (2022): BORNH 03/2022

Here we report the results of a longitudinal field study (2002 -2018) aimed at monitoring the presence and the breeding activity of the Italian Green Toad, Bufotes balearicus, within the area of the administrative boundaries of Naples. We identified breeding sites in five sub-areas (Bagnoli – ex sito industriale, Mostra d'Oltremare, Parco del Poggio, Scalo Ferroviario-zona orientale, Parco Massimo Troisi), one of which (Parco M. Troisi) since 2018. Only in two subareas (Mostra d’Oltremare and Parco del Poggio) the reproductive activity took place regularly, almost every year, although the development of tadpoles was often compromised by a variety of anthropogenic disturbances. However, the major potential threat to the survival of this amphibian for the entire study area is represented by the isolation of its population mainly due to the scarcity of natural and semi-natural green areas in a highly urbanized territory. We hope that our research will be helpful to plan management activity for this species in the urban area of Naples., BORNH Bulletin of Regional Natural History, V. 1 N. 3 (2021): BORNH 03/2021

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.

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.

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

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.

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.

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.

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.

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.

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