Lizards as model organisms of sex chromosome evolution: What we really know from a systematic distribution of available data?

Lizards represent unique model organisms in the study of sex determination and sex chromosome evolution. Among tetrapods, they are characterized by an unparalleled diversity of sex determination systems, including temperature-dependent sex determination (TSD) and genetic sex determination (GSD) under either male or female heterogamety. Sex chromosome systems are also extremely variable in lizards. They include simple (XY and ZW) and multiple (X1X2Y and Z1Z2W) sex chromosome systems and encompass all the different hypothesized stages of diversification of heterogametic chromosomes, from homomorphic to heteromorphic and completely heterochromatic sex chromosomes. The co-occurrence of TSD, GSD and different sex chromosome systems also characterizes different lizard taxa, which represent ideal models to study the emergence and the evolutionary drivers of sex reversal and sex chromosome turnover. In this review, we present a synthesis of general genome and karyotype features of non-snakes squamates and discuss the main theories and evidences on the evolution and diversification of their different sex determination and sex chromosome systems. We here provide a systematic assessment of the available data on lizard sex chromosome systems and an overview of the main cytogenetic and molecular methods used for their identification, using a qualitative and quantitative approach

New data on distribution of amphibians and reptiles in the Aral Sea Basin and surrounding areas of Kazakhstan. Part I. The Green Toads of Bufo viridis complex (Amphibia: Anura)

First the data on distribution of the toads of Bufo viridis complex are presented for Aral Sea Basin and surrounding areas of Southern and Western Kazakhstan. Karyological analysis revealed a presence of diploid toad populations in the valley of the middle and low flows of Syr-Darya River; in the northern coast of Aral Sea and the desert areas between the last and Irgyz-Turgay Basin; in the northern part of Karatau Range. The only tetraploid population was discovered in the central part of Karatau Range

Comparative cytogenetics of Hemorrhois hippocrepis and Malpolon monspessulanus highlights divergent karyotypes in Colubridae and Psammophiidae (Squamata: Serpentes)

Despite the growing interest in the evolutionary cytogenetics of squamates, chromosomal data are lacking for most taxa. We performed a preliminary molecular taxonomic analysis and a comparative cytogenetic study on Hemorrhois hippocrepis and Malpolon monspessulanus. We used a combination of standard karyotyping, chromomycin A3/Methyl green staining, C-banding, Ag-NOR staining and NOR-FISH to provide the first karyotype description of H. hippocrepis and a re-description of the karyotype of M. monspessulanus, including chromosome markers, heterochromatin patterns and sex chromosome systems. Our results show that H. hippocrepis has 2 n = 36 chromosomes, with 16 macro- and 20 microchromosomes and NORs on the 6th pair. The 4th pair represents homomorphic (metacentric) ZW sex chromosomes, but the W chromosome is completely heterochromatic. Malpolon monspessulanus has 2 n = 44 chromosomes, with 20 macro- and 24 microchromosomes, NORs on the 6th telocentric pair. The 4th pair represents the sex chromosomes (ZZ/ZW), with a W chromosome smaller than the Z and completely heterochromatic. Comparing our cytogenetic data to those available from the literature, we note the occurrence and distribution of primitive and derived chromosomal characteristics and discuss the chromosome diversification in two snake clades belonging to Colubridae and Psammophiidae, respectively. We highlight that these two families followed different chromosome diversification pathways, characterised by a highly conserved karyotype structure in Colubridae and a higher chromosome variability in Psammophiidae, mostly driven by a progressive reduction of the chromosome number by means of chromosome fusions. We also provide cytotaxonomic insights supporting the distinction between M. monspessulanus and M. insignitus

Comparative cytogenetics on Zamenis lineatus and Elaphe quatuorlineata (Serpentes: Colubridae)

Because of their peculiar genomic and chromosomal characteristics, reptiles are extraordinary model organ-isms to study karyotype and sex chromosome evolution, but despite the growing interest in their evolutionary cytoge-netics, only a small fraction of species have a known karyotype. We performed a comparative cytogenetic analysis on Elaphe quatuorlineata and Zamenis lineatus, using classic and molecular techniques. We provide the karyotype of these two species and an assessment of their chromosomal features. Chromosome analysis was performed with standard kar-yotyping, C-banding, sequential C-banding + CMA3 + DAPI and Ag-NOR staining. On E. quatuorlineata, we also per-formed CMA3-methyl green staining and Fluorescence in situ Hybridization mapping NOR loci (NOR-FISH). Elaphe quatuorlineata and Z. lineatus show a very similar karyotype of 2n = 36, with 8 macro- and 10 microchromosome pairs, but differ in the morphology of the pair 8, which resulted submetacentric in the former and metacentric in the latter species. By comparing our data to those available from the literature on congeneric species, we analysed the occurrence of primitive and derivate chromosomal characters and provide cytotaxonomic insights, which further support the spe-cies status of Z. lineatus. In both species, the 4th pair was identified as the sex chromosome pair (ZZ/ZW) and NORs were localized on a microchromosome pair. We finally highlight in both genera Elaphe and Zamenis different stages of heterochromatinization of the W chromosome, in agreement with the progressive diversification model of sex chromo-some as already shown in different reptile taxa

Comparative cytogenetics on eight Malagasy Mantellinae (Anura, Mantellidae) and a synthesis of the karyological data on the subfamily

We performed a molecular and cytogenetic analysis on different Mantellinae species and revised the available chromosomal data on this group to provide an updated assessment of its karyological diver sity and evolution. Using a fragment of the mitochondrial 16S rRNA, we performed a molecular taxo nomic identification of the samples that were used for cytogenetic analyses. A comparative cytogenetic analysis, with Giemsa’s staining, Ag-NOR staining and sequential C-banding + Giemsa + CMA + DAPI was performed on eight species: Gephyromantis sp. Ca19, G. striatus (Vences, Glaw, Andreone, Jesu et Schimmenti, 2002), Mantidactylus (Chonomantis) sp. Ca11, M. (Brygoomantis) alutus (Peracca, 1893), M. (Hylobatrachus) cowanii (Boulenger, 1882), Spinomantis prope aglavei “North” (Methuen et Hewitt, 1913), S. phantasticus (Glaw et Vences, 1997) and S. sp. Ca3. Gephyromantis striatus, M. (Brygoomantis) alutus and Spinomantis prope aglavei “North” have a karyotype of 2n = 24 chromosomes while the other species show 2n = 26 chromosomes. Among the analysed species we detected differences in the number and position of telocentric elements, location of NOR loci (alternatively on the 6th, 7th or 10th pair) and in the distribution of heterochromatin, which shows species-specific patterns. Merging our data with those previously available, we propose a karyotype of 2n = 26 with all biarmed elements and loci of NORs on the 6th chromosome pair as the ancestral state in the whole family Mantellidae. From this putative ancestral condition, a reduction of chromosome number through similar tandem fusions (from 2n = 26 to 2n = 24) occurred independently in Mantidactylus Boulenger, 1895 (subgenus Brygoomantis Dubois, 1992), Spinomantis Dubois, 1992 and Gephyromantis Methuen, 1920. Similarly, a relocation of NORs, from the putative primitive configuration on the 6th chromosome, occurred independently in Gephyroman tis, Blommersia Dubois, 1992, Guibemantis Dubois, 1992, Mantella Boulenger, 1882 and Spinomantis. Chromosome inversions of primitive biarmed elements likely generated a variable number of telocentric elements in Mantella nigricans Guibé, 1978 and a different number of taxa of Gephyromantis (subgenera Duboimantis Glaw et Vences, 2006 and Laurentomantis Dubois, 1980) and Mantidactylus (subgenera Brygoomantis, Chonomantis Glaw et Vences, 1994, Hylobatrachus Laurent, 1943 and Ochthomantis Glaw et Vences, 1994)

Karyological diversification in the genus lyciasalamandra (Urodela: Salamandridae)

We performed the first cytogenetic analysis on five out of the seven species of the genus Lyciasalamandra, including seven subspecies, and representatives of its sister genus Salamandra. All the studied species have a similar karyotype of 2n = 24, mostly composed of biarmed elements. C-bands were observed on all chromosomes, at centromeric, telomeric and interstitial position. We found a peculiar taxon-specific NOR configuration, including either heteromorphic and homomorphic NORs on distinct regions of different chromosomes. Lyciasalamandra a. antalyana and L. helverseni showed two homomorphic NORs (pairs 8 and 2, respectively), while heteromorphic NORs were found in L. billae (pairs 6, 12), L. flavimembris (pairs 2, 12), L. l. luschani (pairs 2, 12), L. l. basoglui (pairs 6, 12), L. l. finikensis (pairs 2, 6) and S. lanzai (pairs 8, 10). Homomorphic NORs with an additional supernumerary site were shown by S. s. salamandra (pairs 2, 8) and S. s. gigliolii (pairs 2, 10). This unexpected highly variable NOR configuration is probably derived from multiple independent NOR translocations and paracentric inversions and correlated to lineage divergence in Lyciasalamandra. These results support the taxonomic validity of the studied taxa and are consistent with a hypothesized scenario of synchronous evolution in the genus

Chromosome diversity and evolution in helicoide a (Gastropoda: Stylommatophora): A synthesis from original and literature data

We performed a molecular and a comparative cytogenetic analysis on different Helicoidea species and a review of all the available chromosome data on the superfamily to provide an updated assessment of its karyological diversity. Standard karyotyping, banding techniques, and Fluorescence in situ hybridization of Nucleolus Organizer Region loci (NOR-FISH) were performed on fifteen species of three families: two Geomitridae, four Hygromiidae and nine Helicidae. The karyotypes of the studied species varied from 2n = 44 to 2n = 60, highlighting a high karyological diversity. NORs were on a single chromosome pair in Cernuella virgata and on multiple pairs in four Helicidae, representing ancestral and derived conditions, respectively. Heterochromatic C-bands were found on pericentromeric regions of few chromosomes, being Q-and 4′,6-diamidino-2-phenylindole (DAPI) negative. NOR-associated heterochromatin was C-banding and chromomycin A3 (CMA3) positive. Considering the available karyological evidence on Helicoidea and superimposing the chromosome data gathered from different sources on available phylogenetic inferences, we describe a karyotype of 2n = 60 with all biarmed elements as the ancestral state in the superfamily. From this condition, an accumulation of chromosome translocations led to karyotypes with a lower chromosome number (2n = 50–44). This process occurred independently in different lineages, while an augment of the chromosome number was detectable in Polygyridae. Chromosome inversions were also relevant chromosome rearrangements in Helicoidea, leading to the formation of telocentric elements in karyotypes with a relatively low chromosome count

First karyological analysis of the endemic malagasy phantom gecko matoatoa brevipes (Squamata: Gekkonidae)

The genus Matoatoa includes two Malagasy endemic species, M. brevipes and M. spannringi. Due to their cryptic behaviour, the two species are known only from a handful of specimens and have been included in few molecular studies. Here we carried out a molecular barcoding analysis using a fragment of the mitochondrial NADH dehy-drogenase subunit 2 (ND2) and the first chromosomal analysis of M. brevipes. The molecular analysis confirmed the identity of the studied samples as M. brevipes. However, the level of genetic divergence (4% uncorrected p-distance) between our samples and other sequences of M. brevipes, suggests previously unrecognised diversity within the spe-cies. The karyotype of M. brevipes is composed of 2n = 34 chromosomes: the first pair is metacentric, while all the other pairs are telocentric and gradually decreasing in length (Arm Number, AN = 36). C-banding revealed little evidence of centromeric heterochromatin, while NOR-associated heterochromatin was found on the telomeres of a medi-um sized telocentric pair. No heteromorphic chromosome pairs were found in the karyotype of the species, suggesting that putative sex chromosomes are at an early stage of differentiation. Karyological comparisons with closely related species were performed with Christinus marmoratus, and representatives of the genera Phelsuma, Ebenavia, Paroedura and Uroplatus. Comparisons across genera suggest that chromosome diversification in this group of geckos probably occurred by means of chromosome fusions and inversions, leading to a reduction of the chromosome number and the formation of biarmed elements in different species

Extended-spectrum beta-lactamase-producing and carbapenemase-producing Enterobacter cloacae ventriculitis successfully treated with intraventricular colistin

We present a case of post-neurosurgical ventriculitis caused by carbapenemase-producing Enterobacter cloacae successfully treated with intraventricular colistin. Enterobacter spp are intrinsically resistant to aminopenicillins, cefazolin, and cefoxitin due to the production of constitutive chromosomal AmpC beta-lactamases. Moreover, extended-spectrum beta-lactamase-producing Enterobacter spp have been identified in the USA and Europe, and carbapenems are considered the drug of choice in these cases. Our isolate was sensitive only to fosfomycin, tigecycline, and colistin, and 6 days of intravenous colistin had failed to eradicate the infection. This case provides clinical evidence to support the administration of intraventricular colistin in such patient

Skeletochronology, age at maturity and cause of mortality of loggerhead sea turtles Caretta caretta stranded along the beaches of Campania (south-western Italy, western Mediterranean Sea)

Information on demographic and life-history traits of endangered vertebrate species, such as sea turtles, is crucial for planning management and conservation actions. We applied skeletochronology of phalanges to estimate the age of loggerhead turtles, Caretta caretta, found dead stranded along the beaches of Campania (western Mediterranean) from 2013 to 2017. To obtain maturity data, we examined gonads from a subsample of 7 males and 11 females. Overall, curved carapace length (CCL) ranged from 5.6 to 90.8 cm, but for most turtles (89%) it was 50-79.9 cm. Predominance of stranded females (62%) was recorded. Ten out of the eighteen histologically examined gonads allowed estimating maturity. Based on the lines of arrested growth counting, the estimated age of the examined specimens ranged from 0 (hatchling) to 26 years. The modal age was 14 years for males and 17 years for females. The smallest male with spermatogenetic activity had a CCL of 65 cm and was estimated to be 16 years old. The smallest female with follicular development stage, characterising the transition towards adulthood, had a CCL of 69.5 cm and was estimated to be 20 years old. Anthropogenic factors were responsible for 36% of the mortality of individuals, followed by parasitic/infective pathologies (20%). Copyright Fabio Maria Guarino et al