Sexual isolation between two newt species, Triturus vulgaris and T. montandoni (Amphibia, Urodela, Salamandridae)

In the present study we investigated sexual isolation between Triturus vulgaris and 77 montandoni in mating experiments run under semi-natural conditions. The two newt species offer a suitable model for studying evolution of reproductive isolation and mating preferences because they arc genetically the most similar species within the genus and readily hybridize in nature. Separate experiments were conducted in which groups of virgin females were placed together (in artificial pools) with groups of homospecific, heterospecific or both types of males. The estimates of reproductive isolation and mating propensity were based on the numbers of females producing hybrids and/or non-hybrid progeny. The levels of reproductive isolation, isolation asymmetry (IA) and propensity asymmetry (PA) were significant only for experiments in which females were given a choice between conspecific and heterospecific males. This implies that mating experiments with no interspecific choice may reduce discrimination and affect patterns of IA and PA. Asymmetry in reproductive isolation was also significant when the analysis was confined to just inseminated females. Differences in habitat preferences and condition of females possibly contributed to the relatively high values of PA.     

Heterochromatin characterization of sex chromosomes in Triturus marmoratus (Urodela, Salamandridae).

The sex chromosomes of the Iberian marbled newt, Triturus marmoratus, were studied using various banding techniques, including restriction enzyme/nick translation (RE/NT) procedures. Four types of heterochromatin on the sex chromosomes could be distinguished: (1) distamycin A/DAPI and chromomycin A3/distamycin A positive, EcoRI/NT negative, and HaeIII/NT and HinfI/NT positive; (2) distamycin A/DAPI and chromomycin A3/distamycin A positive, but RE/NT negative; (3) AT rich, but RE/NT negative; and (4) distamycin A/DAPI and chromomycin A3/distamycin A positive, EcoRI/NT and HinfI/NT negative, but HaeIII/NT positive. These data suggest a common origin for the terminal heterochromatic domains of both the X and Y chromosomes in this species.     

Experimental hybridization within the genus Triturus (Urodela: Salamandridae)

The spermatogenesis of 9 F₁ hybrids of Triturus cristatus carnifex × T. vulgaris meridionalis was studied in squash preparations of testicular fragments, treated by the C-staining method. The chromosome number of these hybrids was examined in spermatogonial metaphases and found to be diploid. The two parental sets were always recognized, which means that a regular, although heterospecific, amphimixis occurred (2n=n+n). Meiotic prophase I is greatly altered owing to a failure of typical chromosome pairing and chiasma formation. At metaphase I and/or meta-anaphase I, the effects of the hybrid combination of the 2 specific parental sets are clearly visible. Most primary spermatocytes contain only univalents. A few show chromosome associations (bivalents, trivalents and, more rarely, quadrivalent chains) besides univalents. Such associations are of 2 types: (a) intragenomal associations = associations of 2 chromosomes by a terminal (a¹) or subterminal chiasma (a²); (b) intergenomal associations = associations of 2 chromosomes by a terminal (b¹) or subterminal chiasma (b²). Univalents segregate at random while the associations often lag on the equatorial plane or migrate entire to a spindle pole. Primary spermatocytes with chromosome multivalents can encounter greater difficulties in accomplishing the first cytokinesis. Secondary spermatocytes are numerically and qualitatively unbalanced; however, some of them undergo spermiogenesis and can give rise to a small number of sperms, generally abnormal and never united in bundles. — Problems related to the occurrence of anomalous chiasmata and of intra- and inter-genomal homologies are discussed.     

Experimental hybridization within the genus Triturus (Urodela: Salamandridae)

Spermatogenesis in the F₁ hybrid (2n=24=12 +12 ) between the closely related newt species T. cristatus carnifex and T. marmoratus was apparently normal up to pachytene. Many unpaired chromosomes were present at diplotene and a typical diakinesis was lacking. Primary spermatocytes at meta-and meta-anaphase contained up to 12 regular intergenomal bivalents and a corresponding number of univalents when less then 12 II. Most chiasmata were terminal or subterminal, some intercalary. Chiasmata between corresponding heterospecific chromosomes can be reported as true: real crossing over has taken place, proving the presence of primary chromosomal homologies between the 2 sets of the parental species. Evidence for recombination is based on the segregation of particular markers (i.e., subterminal C-bands and NORs) observed in certain chromosomes at metaphase II. One chromatid of single chromosomes can show the T. cristatus phenotype and the other the T. marmoratus phenotype. A few primary spermatocytes contain a certain number of irregular associations (intragenomal or intrahaploid bivalents, irregular intergenomal bivalents, chromosome multivalents) joined by chiasmata which can be defined as anomalous. Other abnormalities concern the occurrence of interlocked bivalents which occasionally show an anomalous exchange between heterologous chromatids. — Cytogenetic criteria useful to evaluate the taxonomic relationships between different species have been discussed as well as some possible trends in chromosome evolution and speciation within the genus Triturus.     

On the biological significance of certain morphological characters in males of the Smooth newt Triturus vulgaris and of the Palmate newt Triturus helveticus (Urodela: Salamandridae)

The males of Triturus vulgaris and Triturus helveticus can be differentiated by a number of morphological characters whose biological function is unknown. It is suggested that these characters enhance the effectiveness of male displays performed during sexual behaviour. This hypothesis is discussed in relation to the taxonomic and genetic relationships between the two species.     

Endocrine-disrupting effects of nonylphenol in the newt, Triturus carnifex (Amphibia, Urodela)

The aim of our study was to verify whether environmental concentrations of nonylphenol influenced the adrenal gland of Triturus carnifex. Newts were exposed to 19 μg/L nominal concentration of nonylphenol throughout the periods of December-January and March-April, corresponding to different stages of the chromaffin cell functional cycle. The morphological features of the steroidogenic and chromaffin tissues, and the serum levels of ACTH, aldosterone, corticosterone, norepinephrine and epinephrine were evaluated. Nonylphenol did not influence ACTH serum levels. During the two periods examined, the steroidogenic tissue had the same reaction: the quantity of cytoplasmic lipids, and the corticosteroid serum levels, decreased, suggesting the inhibition of synthesis and release of corticosteroids. During the two periods examined, the chromaffin tissue reacted differently to nonylphenol. During December-January, the numeric ratio of norepinephrine granules to epinephrine granules, and the epinephrine serum levels, increased, suggesting the stimulation of epinephrine release. During March-April, the numeric ratio of norepinephrine granules to epinephrine granules did not change, and the norepinephrine serum levels decreased, suggesting the inhibition of norepinephrine release. Our results show that nonylphenol influences the activity of the newt adrenal gland; considering the physiological role of this gland, our results suggest that nonylphenol may contribute to amphibian decline.     

Asymmetric female preferences for courtship pheromones in two closely-related newt species, the smooth newt (Lissotriton vulgaris) and the Carpathian newt (L. montandoni) (Salamandridae)

The smooth (Lissotriton vulgaris) and Carpathian (L. montandoni) newts are sister species. These are separated by a moderate genetic distance, but exhibit striking morphological differences, especially in male epigamic traits. In the areas where they co-occur, they readily mate with each other and produce viable hybrids. However, a high level of pre-zygotic isolation with an unknown behavioral basis has been reported. The complex courtship of newts consists of at least three types of modality: chemical, visual, and tactile. The relative significance of these in mate choice is unclear, but it is commonly accepted that pheromones are an important communication channel. The goal of this study was to determine whether the females of L. vulgaris and L. montandoni exhibit preferences for conspecific extracts from the pheromone-producing abdominal (dorsal) glands. Females of both species spent more time in proximity to the source of the abdominal gland extracts of their own species when a liver extract was presented as an alternative. In a second trial, females were simultaneously confronted with conspecific and heterospecific abdominal gland extracts. Asymmetric preferences were found. Lissotriton vulgaris females were not selective, whereas L. montandoni females preferred the conspecific abdominal gland extract. This finding is consistent with the results of earlier experiments on mate choice in these species. The results strongly indicate that pheromones play a crucial role in courtship and species recognition in this pair of closely related, hybridizing species.     

Sperm transport after insemination in the Alpine newt (Triturus alpestris, Caudata, Salamandridae)

The goal of this study was to test if sperm transport to the spermathecae in the Alpine newt (Triturus alpestris) requires active co-operation of the female. Artificial insemination of anaesthetised female newts was conducted using spermatophores collected from courting males and with sperm duct contents collected from sacrificed males. Sperm was present in the spermathecae of 9 out of 10 females inseminated with the spermatophores but in only 1 out of 8 females inseminated with sperm duct contents. The females of both groups laid some eggs after insemination, and a portion of these eggs in group of females inseminated with spermatophores were fertilized. However, the number of eggs produced by the females was much lower than typical egg-production in newts. The presence of sperm in the spermathecae of females inseminated with spermatophores and lack of sperm in the spermathecae of females inseminated with sperm duct contents suggests that sperm transport is either induced by the substances present in spermatophores and/or that sperm from the sperm duct is not fully mobile in comparison with sperm from the spermatophores.     

Differential staining and chromatin packing of the mitotic chromosomes of the newt Triturus cristatus

Mitotic metaphase chromosomes of cold-treated Triturus cristatus show a characteristic pattern of constrictions, most of which lie close, though not immediately adjacent, to the centromeres. The chromatin in these cold-induced constrictions stains intensely with Giemsa. Cold-treated spermatogonia show spiral structure throughout the metaphase chromatids; the packing of chromatin fibrils is much tighter in the constricted regions than elsewhere, and the gyres in the constricted regions are narrower and of shorter pitch.     

A parallel geographical mosaic of morphological and behavioural aposematic traits of the newt, Cynops pyrrhogaster (Urodela: Salamandridae)

Aposematic animals advertise their unprofitability to potential predators with conspicuous coloration, occasionally in combination with other life-history traits. Theory posits that selection on functionally interrelated aposematic characters promotes the unidirectional evolution of these characters, resulting in an increase or decrease in the effectiveness of the signal. To test whether this prediction applies on a microevolutionary scale, the intra- and interpopulational variations in aposematic coloration, behaviour (which enhances the effectiveness of the coloration) and body size of newts, Cynops pyrrhogaster (Urodela: Salamandridae), were investigated. A parallel geographical mosaic of variation in aposematic coloration and behaviour among populations, independent of body size, was found. Newts on islands displayed more conspicuous aposematic traits than those on the mainland, both morphologically and behaviourally. There was no significant relationship between variation in coloration and behaviour within populations. Male newts displayed more conspicuous coloration than females. Surveys of potential predators suggest that variable natural selection at a local scale, such as predation pressure, may primarily be responsible for the microevolution of variable aposematic traits in newts.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 613–622.     

Sexual Interference and Sexual Defense in the Smooth Newt,Triturus vulgaris (Amphibia,Urodela, Salamandridae)

• 1 When a male smooth newt encounters a ♀ who is already engaged in courtship, he may mimic her behaviour during the spermatophore deposition and transfer stages of the courtship. He thereby usurps the courting ♂ and may inseminate the ♀ himself. Such sexual interference depresses the short-term, and perhaps long-term, mating success of the courting ♂.
• 2 In the presence of a potential rival, the courting ♂ alters certain aspects of his sexual behaviour. He displays more intensely to the ♀ and attempts to draw her away from the rival by increasing the duration of his display. He may also “check” that it is the ♀, and not the rival, who will elicit the deposition of a spermatophore from him. These changes in the behaviour of the courting ♂ are interpreted as defense against sexual interference.
• 3 Female smooth newts may be multiply inseminated as a consequence of sexual interference; this may result in sperm competition. However, ♀♀ seem to find competitive interactions between ♂ ♂ “aversive”.
• 4 Sexual interference by ♀-mimicry and associated defensive behaviour patterns are common in the urodele amphibians. Interference can be thought of as a “side-payment” conditional mating strategy.

     

The sexual behaviour of the red-spotted newt, Notophthalmus viridescens (Amphibia: Urodela: Salamandridae)

The sexual behaviour of the red-spotted newt, Notophthalmus viridescens is described. The male can adopt one of two types of behaviour depending on the initial responsiveness of the female. If she is responsive, the male performs a brief lateral (or hula) display, at the end of which one or more spermatophores may be deposited. If she is unresponsive, the male captures the female and a lengthy period of amplexus ensues, followed by spermatophore transfer behaviour. Spermatophore transfer is most successful after amplexus, and a preliminary cost-benefit analysis of the two types of courtship is given. The behaviour is discussed in the context of other urodele amphibians, and in the wider context of male reproductive strategies.     

The Cloaca and Spermatheca of the Female Smooth Newt, Triturus vulgaris L. (Amphibia: Urodela: Salamandridae)

The anatomy and histology of the cloaca and spermatheca of 13 female smooth newts ( Triturus vulgaris ) were studied by light microscopy. The cloaca consists of a short, anterior tube (which receives the oviducts), which opens into a larger, more posterior chamber. The spermatheca, which is the only gland in the cloaca, consists of a mass of exocrine, acinar tubules which empty individually and dorsolaterally into the posterior part of the cloacal tube and all but the most posterior part of the cloacal chamber. Stored sperm are most abundant in the spermathecal tubules during the peak of the breeding season (in May and June); during their period of storage, the sperm do not appear to make intimate contact with the epithelial cells lining the spermathecal tubules. Longterm storage of viable sperm from one breeding season to the next is probably absent in the smooth newt. Sperm storage by and multiple insemination of females both occur in this species and are necessary preconditions for competition between the sperm of different males for the fertilization of eggs. However, sperm competition has not been demonstrated in the smooth newt.     

Field observations of the sexual behaviour of the smooth newt, Triturus vulgaris vulgaris (Amphibia: Salamandridae)

This study reports observations of smooth newts, Triturus vulgaris vulgaris , breeding in a pond in southern England. Efforts were made to collect data on the timing of oviposition and on sexual behaviour during the breeding seasons of 1985 and 1986. Despite some between-year differences in the timing of certain events, the qualitative pattern of reproduction was similar in both years. Courtship interactions were seen most frequently during a relatively short period of time before females began laying their eggs, which occurred in a highly synchronized manner. It seems likely that the probability of any one courtship encounter resulting in spermatophore transfer was low. Males competed for mates by chasing females and by interfering with one another's courtships, but no overt aggression was seen. Females appeared to find such male-male interactions 'aversive'. Scramble competition between males was most intense when the majority of females in the population were laying their eggs and were unresponsive to courtship. The mating system of the smooth newt most closely resembles a lek in which the intensity of sexual selection among males varies as a function of female availability.     

Brain-capillary architecture of the newt,Triturus cristatus carnifex (Caudata: Salamandridae): Scanning electron-microscopical study of vascular corrosion casts

The brain-microvascular architecture of the newt (Triturus cristatus carnifex) was studied by scanning electron-microscopical examination of microvascular corrosion casts. It is characterized at the leptomeningeal level by a superficial network of single vessels that leave the meningeal sheath to form hairpin-shaped capillary loops. These loops penetrate the nervous parenchyma to varying depths and are variously bent, inclined, and twisted. Each hairpin-like loop consists of descending and ascending limbs connected by a distal U-shaped segment. The limbs are of equal diameter and lack physical differentiation. Some loops branch in a secondary loop, but there are no intraparenchymal connections with analogous surrounding structures. On vascular casts we observed detailed imprints of endothelial cell nuclei. This microvascular architecture is present in the whole brain except in leptomeninx and choroidal plexuses. The vascular pattern of T. cristatus carnifex resembles that of apodans. However, it differs from the anuran condition, which is characterized by a three-dimensional parenchymal network and from that of Ambystoma mexicanum which displays an intermediate condition between a single-capillary network and a capillary-loop model. At the level of paired capillaries, a counter-current exchange system may occur between the two limbs of the same loop, an arrangement that could promote uniformity of extracellular fluid composition in parenchyma.     

Breeding dynamics of Triturus carnifex at a pond in northwestern Italy (Amphibia, Urodela, Salamandridae)

A population of the Italian crested newt ( Triturus carnifex ) was studied in 1984 at a pond near Druento (Turin, Piedmont, northwestern Italy). Animals entering and leaving the pond fell into paired pit-fall traps during both spring and autumn migrations. A positive correlation found between rainfall and migration, a shortening of migration periods and summer inactivity suggest that, in the Mediterranean region, air moisture may be a limiting factor for newt activity. During their aquatic period adults showed strong individual differences in weight changes that cannot be explained on the basis of initial body weight or length. High variability in crested newt life history traits may be the result of repeated bottlenecks caused by environmental stress and high population isolation.     

Natural variation in the limb skeletal pattern of the crested newt,Triturus carnifex (amphibia: Salamandridae)

The forelimb skeletal patterns of two geographically isolated populations of Triturus carnifex, one from northern Italy (Rosate, Milano) and one from central Italy (Bagnaia, Perugia) were studied. A total of 1,018 limbs were examined. Limb skeletal variants, generally consisting either of a reduced number of carpals and/or of an alternate phalangeal formula, were commonly observed, as has been reported for other amphibian species. The population from northern Italy showed a greater variability than that from central Italy. Skeletal variants were present in 34% and 12% of the limbs, respectively (i.e., in 48% and 20% of the newts). In the Rosate population, the majority of variants consisted of fusions of radiale and prepollicis and of the phalangeal formula 1-2-3-2. Severe skeletal defects were also found in 3% of the limbs of the newts from Rosate and in 1% of the limbs of those from Bagnaia. At present, no conclusion can be drawn about the evolutionary significance of the limb skeletal variation observed in the two populations, because of the paucity of data on intra- and interpopulation variations in other species of the genus Triturus. © 1992 Wiley-Liss, Inc.