Complex ontogeny of sexual size dimorphism in a female-larger gecko: Implications of determinate growth for lizard body size and life-history evolution.

Ectothermic vertebrates such as reptiles were assumed to be indeterminate growers, which means that there is no terminal point in time or size for growth in their lifetime. In recent years, evidence for the determinate nature of growth in lizards has accumulated, necessitating a re-examination of models of their ontogeny and evolution of sexual size dimorphism (SSD). In the female-larger gecko Paroedura vazimba, we monitored post-embryonic growth over a period of 15 months. After hatching, females grew faster than males but also reached their final body size, that is, closed growth of their vertebrae, earlier than males. The closure of bone growth in females correlates with the onset of reproductive maturation. We compared this pattern with the previously minutely studied, male-larger species Paroedura picta, where we documented determinate growth as well. We propose a model to explain the evolutionary switches in the direction of SSD in lizards based on bipotential effects of ovarian hormones on growth. In this model, male growth is assumed to require no male-specific growth modifier, such as sex-limited hormonal regulators, while growth is feminized by ovarian hormones in females. Low levels of ovarian hormones can promote bone growth, but high levels associated with maturation of the reproductive organs promote senescence of bone growth plates and thus cessation of bone growth. We suggest that models on growth, life-history and evolution of body size in many lizards should acknowledge their determinate nature of growth.

Two transcriptionally distinct pathways drive female development in a reptile with both genetic and temperature dependent sex determination.

How temperature determines sex remains unknown. A recent hypothesis proposes that conserved cellular mechanisms (calcium and redox; 'CaRe' status) sense temperature and identify genes and regulatory pathways likely to be involved in driving sexual development. We take advantage of the unique sex determining system of the model organism, Pogona vitticeps, to assess predictions of this hypothesis. P. vitticeps has ZZ male: ZW female sex chromosomes whose influence can be overridden in genetic males by high temperatures, causing male-to-female sex reversal. We compare a developmental transcriptome series of ZWf females and temperature sex reversed ZZf females. We demonstrate that early developmental cascades differ dramatically between genetically driven and thermally driven females, later converging to produce a common outcome (ovaries). We show that genes proposed as regulators of thermosensitive sex determination play a role in temperature sex reversal. Our study greatly advances the search for the mechanisms by which temperature determines sex.

Is the natural UV zone important for successful captive propagation of the Panther Chameleon (Furcifer pardalis); are different UVB irradiance exposures that generate a similar dose equally successful?

While we generally understand the optimal ultraviolet B (UVB) environment for the growth and reproduction of female Panther Chameleons Furcifer pardalis, we do not know the relative importance of UVB irradiance and dose for optimal husbandry outcomes. Accordingly, we experimented with Panther Chameleon females to test the hypothesis that UVB dose (irradiance × exposure duration) determines the outcome, regardless of the combination of UVB irradiance and exposure duration generating the dose. We varied UVB irradiance and exposure duration across treatment groups while keeping dose similar and within a range previously documented to result in reproductive success. The growth rate, age of maturity, and measurable vitamin D status were not significantly different among the treatment groups. Individuals in all groups produced viable eggs that successfully hatched. Thus, we found some support for the hypothesis that the UVB dose determines the outcome regardless of UVB irradiance. However, mean egg vitamin D3 concentration and percent hatching were higher in the highest UVB irradiance group, despite similar doses among the three groups. Preliminary field data reveal that this species occupies UV irradiance Zone 4 in Madagascar, the highest zone for reptiles recorded. Only the irradiance of the high UVB irradiance group in our experiment approached this zone and resulted in the best reproductive success. Biosynthesis of vitamin D3 and provisioning to eggs is more efficient when exposure to UVB irradiance is similar to that in their natural environment. Establishing an optimal UVB environment, based on knowledge of the natural UVB environment, is important for the propagation of Panther Chameleons in captivity.

Transcriptomic and proteomic analysis of Hemidactylus frenatus during initial stages of tail regeneration.

Epimorphic regeneration of appendages is a complex and complete phenomenon found in selected animals. Hemidactylus frenatus, house gecko has the remarkable ability to regenerate the tail tissue upon autotomy involving epimorphic regeneration mechanism. This study has identified and evaluated the molecular changes at gene and protein level during the initial stages, i.e., during the wound healing and repair mechanism initiation stage of tail regeneration. Based on next generation transcriptomics and De novo analysis the transcriptome library of the gecko tail tissue was generated. A total of 254 genes and 128 proteins were found to be associated with the regeneration of gecko tail tissue upon amputation at 1, 2 and 5-day post amputation (dpa) against control, 0-dpa through differential transcriptomic and proteomic analysis. To authenticate the expression analysis, 50 genes were further validated involving RTPCR. 327 genes/proteins identified and mapped from the study showed association for Protein kinase A signaling, Telomerase BAG2 signaling, paxillin signaling, VEGF signaling network pathways based on network pathway analysis. This study empanelled list of transcriptome, proteome and the list of genes/proteins associated with the tail regeneration.

Morphospace analysis leads to an evo-devo model of digit patterning.

Biological forms occupy a constrained portion of theoretical morphospaces. Developmental models accounting for empirical morphospaces are necessary to achieve a better understanding of this phenomenon. We analyzed the phalangeal formulas (PFs) in lizards and relatives' hands by comparing them with a set of simulated PFs that compose a theoretical morphospace. We detected that: (1) the empirical morphospace is severely limited in size, (2) the PFs comply with two properties of phalangeal count per digit, namely the ordering rule (DI ≤ DII ≤ DIII ≤ DIV ≥ DV), and the contiguity relationship (neighbor digits differ on average in one phalanx), (3) the totality of the PFs can be categorized into four categories of hands aligned along a feasibility gradient. We also reconstructed the evolution of PFs and found a stepwise trajectory from the plesiomorphic PF towards reduced conditions. Finally, we propose a developmental model as the generative mechanism behind the PFs. It is consistent with the bulk of evidence managed and involves an ordered digit primordia initialization timed with periodic signals of joint formation coming from digit tips. Our approach is also useful to address the study of other meristic sequences in nature such as dental, floral, and branchial formulas.

Maternal Temperature, Corticosterone, and Body Condition as Mediators of Maternal Effects in Jacky Dragons (Amphibolurus muricatus).

AbstractTemperature is a crucial environmental component that imposes physiological constraints and ultimately produces variation in life-history traits. Temperatures experienced by mothers can influence offspring phenotypes, including growth and sex ratios, especially in ectothermic species. However, mechanisms by which thermal information can be passed onto offspring have been underexplored. Here, we investigated corticosterone as a potential mediator of thermal maternal effects. We held female jacky dragons (Amphibolurus muricatus) in two different thermal regimes (short [7 h] or long [11 h] basking treatments), then quantified plasma corticosterone levels and tested for correlations between the resulting corticosterone levels and reproductive outputs. Lizards in the long-bask treatment had significantly higher corticosterone levels than those in the short-bask treatment. Maternal corticosterone, in turn, had sex-dependent effects on offspring hatching size but was not associated with maternal reproductive effort or offspring sex or growth. In contrast, maternal body condition was strongly positively related to both reproductive output (including clutch size and total number of eggs) and offspring size at hatching but had no effect on offspring growth. Basking treatment also interacted with condition and corticosterone to affect egg mass and hatchling snout-vent length, respectively. When we tested for relationships between corticosterone levels and body condition, we found corticosterone to be negatively related to condition in long-bask lizards but only in the postbreeding season. These findings indicate that thermal opportunity alters physiology, with potential consequences for fitness. Moreover, the results suggest interactive influences of temperature, corticosterone, and condition as mediators of maternal effects.

Reproductive cycle and maturation of Swinhoe's tree lizard (Diploderma swinhonis (Günther, 1864)) in Hyuga City, Miyazaki Prefecture, Japan.

Swinhoe's tree lizard (Diploderma swinhonis) is an arboreal agamid that is native to Taiwan. The species has been introduced to some areas of Japan and is regarded as an invasive alien species. In 2016, a nonnative population of D. swinhonis was discovered in Hyuga City, Miyazaki Prefecture, Japan, but little information was available on the ecology of the population at the time. The main purpose of this study was therefore to investigate the reproductive cycle and maturation of this population. Field research was conducted from 2017 to 2019, and 764 lizards were collected. Euthanized lizards were dissected and the reproductive organs were examined to determine the reproductive period, clutch size, clutch frequency and size at sexual maturity. Females with oviductal eggs or vitellogenic ovarian follicles were observed from May to October. Clutch size ranged from 2 to 8, and clutch frequency was more than twice a year. In males, spermiogenesis started in early May and testicular regression was observed in September. Males with spermatozoa in the epididymides were found from May to November. Minimum snout-vent length at sexual maturity was 50.2 mm in females and 53.0 mm in males. Comparisons of the findings of this study and reports from Taiwan suggest that the nonnative population of D. swinhonis in Hyuga City has a higher fecundity than populations in Taiwan. It is therefore considered necessary to exterminate the population in Hyuga City before this species colonizes other areas.

Effect of early thermal environment on the morphology and performance of a lizard species with bimodal reproduction.

Early developmental environments influence life-history traits and survival of reptiles. In fact, rising global temperatures have already caused widespread extinctions among lizards. Viviparous species might be more susceptible to increasing temperatures because of their inability to meet their energetic demands following rapid environmental changes. Nevertheless, we do not yet fully understand how lizards with different reproductive modes can respond to climate change. We investigated the effect of both maternal thermal environment during pregnancy and incubation temperature on hatchling morphology and physiological performance of two populations of the lizard Saiphos equalis differing in their mode of reproduction, to test whether reproductive mode affects the ability to buffer against rising temperatures. Gravid females from both populations were subjected to current or projected end-of-century (future) thermal environments, to evaluate differences in the body size, growth rate, thermal preference, and locomotor performance of their offspring. Our results show that independently of the mode of reproduction, high temperatures accelerated gestation periods. Thermal environments did not affect hatchling thermal preference, but viviparous hatchlings consistently preferred lower temperatures. Unlike viviparous lizards, oviparous hatchlings incubated under future temperatures were smaller and had a lower growth rate compared to current-incubated hatchlings. Finally, thermal environments did not affect hatchling endurance and speed when controlling for body size. Our results show that global warming is likely to have a negative impact on S. equalis, but suggest that some of its effects may be ameliorated by maternal responses during pregnancy, particularly in viviparous populations.

Gene expression of the IGF hormones and IGF binding proteins across time and tissues in a model reptile.

The insulin and insulin-like signaling (IIS) network regulates cellular processes including pre- and postnatal growth, cellular development, wound healing, reproduction, and longevity. Despite their importance in the physiology of vertebrates, the study of the specific functions of the top regulators of the IIS network, insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs), has been mostly limited to a few model organisms. To expand our understanding of this network, we performed quantitative gene expression of IGF hormones in liver and qualitative expression of IGFBPs across tissues and developmental stages in a model reptile, the brown anole lizard (Anolis sagrei). We found that lizards express IGF2 across all life stages (preoviposition embryos to adulthood) and at a higher level than IGF1, which is opposite to patterns seen in laboratory rodents but similar to those seen in humans and other vertebrate models. IGFBP expression was ubiquitous across tissues (brain, gonad, heart, liver, skeletal muscle, tail, and regenerating tail) in adults, apart from IGFBP5, which was variable. These findings provide an essential foundation for further developing the anole lizard as a physiological and biomedical reptile model, as well as expanding our understanding of the function of the IIS network across species.

Do all geckos hatch in the same way? Histological and 3D studies of egg tooth morphogenesis in the geckos Eublepharis macularius Blyth 1854 and Lepidodactylus lugubris Duméril & Bibron 1836.

The egg tooth of squamates evolved to facilitate hatching from mineralized eggshells. Squamate reptiles can assist their hatching with a single unpaired egg tooth (unidentates) or double egg teeth (geckos and dibamids). Egg tooth ontogeny in two gekkotan species, the leopard gecko Eublepharis macularius and the mourning gecko Lepidodactylus lugubris, was compared using microtomography, scanning electron microscopy, and light microscopy. Investigated species are characterized by different hardnesses of their eggshells. Leopard geckos eggs have a relatively soft and flexible parchment (leathery) shell, while eggshells of mourning geckos are hard and rigid. Embryos of both species, like other Gekkota, have double egg teeth, but the morphology of these structures differs between the investigated species. These differences in shape, localization, and spatial orientation were present from the earliest stages of embryonic development. In mourning gecko, anlagen of differentiating egg teeth change their position on the palate during embryonic development. Initially they are separated by condensed mesenchyme, but later in development, their enamel organs are connected. In leopard geckos, the localization of egg tooth germs does not change, but their spatial orientation does. Egg teeth of this species shift from inward to outward orientation. This is likely related to differences in structure and mechanical properties of eggshells in the studied species. In investigated species, two hatching mechanisms are possible during emergence of young individuals. We speculate that mourning geckos break the eggshell through puncturing action with egg teeth, similar to the pipping phase of chick and turtles embryos. Egg teeth of leopard geckos cut egg membranes similarly to most squamates. Our results also revealed differences in egg tooth implantation between Gekkota and Unidentata: gekkotan egg teeth are subthecodont (in shallow sockets), while those in unidentates are acrodont (attached to the top of the alveolar ridge). © 2020 Wiley Periodicals LLC.

Cross-Species BAC Mapping Highlights Conservation of Chromosome Synteny across Dragon Lizards (Squamata: Agamidae).

Dragon lizards (Squamata: Agamidae) comprise about 520 species in six subfamilies distributed across Asia, Australasia and Africa. Only five species are known to have sex chromosomes. All of them possess ZZ/ZW sex chromosomes, which are microchromosomes in four species from the subfamily Amphibolurinae, but much larger in Phrynocephalus vlangalii from the subfamily Agaminae. In most previous studies of these sex chromosomes, the focus has been on Australian species from the subfamily Amphibolurinae, but only the sex chromosomes of the Australian central bearded dragon (Pogona vitticeps) are well-characterized cytogenetically. To determine the level of synteny of the sex chromosomes of P. vitticeps across agamid subfamilies, we performed cross-species two-colour FISH using two bacterial artificial chromosome (BAC) clones from the pseudo-autosomal regions of P. vitticeps. We mapped these two BACs across representative species from all six subfamilies as well as two species of chameleons, the sister group to agamids. We found that one of these BAC sequences is conserved in macrochromosomes and the other in microchromosomes across the agamid lineages. However, within the Amphibolurinae, there is evidence of multiple chromosomal rearrangements with one of the BACs mapping to the second-largest chromosome pair and to the microchromosomes in multiple species including the sex chromosomes of P. vitticeps. Intriguingly, no hybridization signal was observed in chameleons for either of these BACs, suggesting a likely agamid origin of these sequences. Our study shows lineage-specific evolution of sequences/syntenic blocks and successive rearrangements and reveals a complex history of sequences leading to their association with important biological processes such as the evolution of sex chromosomes and sex determination.

Lizard Embryos Prioritize Posthatching Energy Reserves over Increased Hatchling Body Size during Development.

Embryonic development in oviparous organisms is fueled by maternally allocated yolk, and many organisms hatch before that energy store is used completely; the resultant leftover (residual) yolk is internalized and may support early posthatching life. However, embryos that use most, or all, of their yolk supply before hatching should hatch at a larger size than those that do not exhaust those energy reserves, which could also have benefits for posthatching growth and survival. To examine the trade-off between residual yolk and offspring size, we experimentally reduced yolk quantity at oviposition in lizard eggs (Amphibolurus muricatus) and then quantified offspring size and the amount of internalized residual yolk. This design enabled us to determine whether embryos (1) exhaust yolk supply during development (thereby maximizing neonatal size) or (2) reduce neonatal size by retaining yolk reserves at hatching. Our data support the latter scenario. Eggs from the yolk-reduced treatment produced smaller offspring with a proportion of residual yolk similar to that of offspring from unmanipulated eggs, suggesting that the fitness benefits of posthatching energy stores outweigh those of larger neonatal size.

Transcriptome sequencing reveals signatures of positive selection in the Spot-Tailed Earless Lizard.

The continual loss of threatened biodiversity is occurring at an accelerated pace. High-throughput sequencing technologies are now providing opportunities to address this issue by aiding in the generation of molecular data for many understudied species of high conservation interest. Our overall goal of this study was to begin building the genomic resources to continue investigations and conservation of the Spot-Tailed Earless lizard. Here we leverage the power of high-throughput sequencing to generate the liver transcriptome for the Northern Spot-Tailed Earless Lizard (Holbrookia lacerata) and Southern Spot-Tailed Earless Lizard (Holbrookia subcaudalis), which have declined in abundance in the past decades, and their sister species, the Common Lesser Earless Lizard (Holbrookia maculata). Our efforts produced high quality and robust transcriptome assemblies validated by 1) quantifying the number of processed reads represented in the transcriptome assembly and 2) quantifying the number of highly conserved single-copy orthologs that are present in our transcript set using the BUSCO pipeline. We found 1,361 1-to-1 orthologs among the three Holbrookia species, Anolis carolinensis, and Sceloporus undulatus. We carried out dN/dS selection tests using a branch-sites model and identified a dozen genes that experienced positive selection in the Holbrookia lineage with functions in development, immunity, and metabolism. Our single-copy orthologous sequences additionally revealed significant pairwise sequence divergence (~.73%) between the Northern H. lacerata and Southern H. subcaudalis that further supports the recent elevation of the Southern Spot-Tailed Earless Lizard to full species.

Cytogenetic mechanisms of unisexuality in rock lizards.

Darevskia rock lizards is a unique complex taxa, including more than thirty species, seven of which are parthenogenetic. In mixed populations of Darevskia lizards, tri- and tetraploid forms can be found. The most important issues in the theory of reticulate evolution of Darevskia lizards are the origin of parthenogenetic species and their taxonomic position. However, there is little data on how meiosis proceeds in these species. The present work reports the complex results of cytogenetics in a diploid parthenogenetic species - D. unisexualis. Here we detail the meiotic prophase I progression and the specific features оf mitotic chromosomes organization. The stages of meiosis prophase I were investigated by immunocytochemical analysis of preparations obtained from isolated primary oocytes of D. unisexualis in comparison with maternal species D. raddei nairensis. It has been shown that in D. unisexualis at the leptotene-zygotene stages the axial elements and the synaptonemal complex (SC) form typical "bouquets". At the pachytene-diplotene stage, 18 autosomal SC-bivalents and thickened asynapted sex Z and w univalents were observed. The presence of SYCP1 protein between the lateral elements of autosomal chromosomes proved the formation of assembled SCs. Comparative genomic hybridization (CGH) on the mitotic metaphase chromosomes of D. unisexualis was carried out using the genomic DNA isolated from the parental species D. raddei nairensis and D. valentini. In the pericentromeric regions of half of the mitotic chromosomes of D. unisexualis, specific regions inherited from maternal species have been found. Following our results, we suggest a model for diploid germ cells formation from diploid oocytes without premeiotic duplication of chromosomes in the oogenesis of diploid parthenogenetic lizards D. unisexualis. Taken as a whole, our findings confirm the hybrid nature of D. unisexualis and shed light on heterozygosity and automixis in diploid parthenogenetic forms.

Embryonic and post-embryonic responses to high-elevation hypoxia in a low-elevation lizard.

Low-elevation species can migrate toward higher elevations to survive in a warming world. However, animals' responses to hypoxia when migrating to high elevations have rarely been addressed. To identify the response of low-elevation lizards to high-elevation hypoxia, we collected field body temperatures (Tfb ) and operative temperatures (Te ) of lizards (Eremias argus) from a low-elevation population (1036 m) and a high-elevation population (2036 m), and then determined adult thermal physiology, embryonic development, and hatchling phenotypes after acclimating low-elevation lizards and incubating their eggs in conditions mimicking the low-elevation oxygen condition (18.5% O2 ) and high-elevation oxygen (hypoxic) condition (16.5% O2 ). Our study revealed that Tfb and Te were higher for the low-elevation population compared to the high-elevation population. We also found adults from low elevation acclimated to hypoxia preferred lower body temperatures, but did not show changes in locomotor performance or growth. In addition, hypoxia did not affect embryonic development (hatching time and success) or hatchling phenotypes (body size and locomotor performance). These results suggest that adult lizards from low elevations can respond to hypoxia-induced stress when migrating to high elevations by behaviorally thermoregulating to lower body temperatures in order to sustain normal functions. Similarly, low-elevation embryos can develop normally (with unchanged hatching success and offspring phenotypes) under the high-elevation hypoxic condition. This study highlights that low-elevation populations of a species that inhabits a range of elevations can buffer the impact of high-elevation hypoxic conditions to some degree and thus attain similar fitness to the source population.

Differential effects of habitat loss on occupancy patterns of the eastern green lizard Lacerta viridis at the core and periphery of its distribution range.

The effects of habitat loss on the distribution of populations are often linked with species specialization degree. Specialist species can be more affected by changes in landscape structure and local patch characteristics compared to generalist species. Moreover, the spatial scale at which different land covers (eg. habitat, cropland, urban areas) affect specialist species can be smaller. Specialization is usually assumed as a constant trait along the distribution range of species. However, for several taxa, there is evidence of higher specialization degree in peripheral populations compared with populations in the core. Hence, peripheral populations should have a higher sensitivity to habitat loss, and strongest effects should be found at a smaller spatial scale. To test these expectations, we implemented a patch-landscape approach at different spatial scales, and compared effects of landscape structure and patch characteristics on occupancy probability among northern peripheral, more specialized populations (Czech Republic) and core populations (Bulgaria) of the eastern green lizard Lacerta viridis. We found that landscape structure and patch characteristics affect differently the occupancy probability of Lacerta viridis in each region. Strongest effects of habitat loss were found at a spatial scale of 150m around patches in the periphery, but at a scale of 500m in the core. In the periphery occupancy probability of populations was principally affected by landscape composition, and the effect of habitat quality was stronger compared to core populations. In the core, persistence of populations was mainly explained by characteristics of the spatial configuration of habitat patches. We discuss possible ecological mechanisms behind the relationship between sensitivity to habitat loss, populations' specialization degree and position in the distribution range, and suggest conservation measures for L. viridis.

Little if any role of male gonadal androgens in ontogeny of sexual dimorphism in body size and cranial casque in chameleons.

Proximate control of the development of sexual dimorphism is still hotly debated in reptiles. In some squamates, many male-typical exaggerated traits including body size were assumed to be controlled by masculinization by male gonadal androgens. We performed a manipulative experiment to test the importance of this mechanism in the development of pronounced sexual differences in body size and size of head casque in the chameleon Chamaeleo calyptratus. Castrated males attained male-typical body size highly deviating from the body size of control females. Ontogenetic allometries of casque size on head length revealed that sexes depart considerably in casque growth later in the ontogeny; however, castrated males still follow male-typical casque growth. Paradoxically, exogenous testosterone led in females to slight increase of casque size, which might reflect interference with the feminizing effects of female gonadal hormones. The results in males strongly suggest that masculinization by male gonadal androgens during growth is not required for the development of sexual dimorphism in body size and casque size in the chameleon. The ontogeny of sexually dimorphic body size and exaggerated traits in at least some squamates is likely controlled by other proximate mechanism, possibly by feminization by ovarian hormones.

Acclimation to Water Restriction Implies Different Paces for Behavioral and Physiological Responses in a Lizard Species.

Chronic changes in climate conditions may select for acclimation responses in terrestrial animals living in fluctuating environments, and beneficial acclimation responses may be key to the resilience of these species to global changes. Despite evidence that climate warming induces changes in water availability, acclimation responses to water restriction are understudied compared with thermal acclimation. In addition, acclimation responses may involve different modes, paces, and trade-offs between physiological and behavioral traits. Here, we tested the dynamical acclimation responses of a dry-skinned terrestrial ectotherm to chronic water restriction. Yearling common lizards (Zootoca vivipara) were exposed to sublethal water restriction during 2 mo of the summer season in laboratory conditions, then released in outdoor conditions for 10 additional months. Candidate behavioral (exploration, basking, and thermal preferences) and physiological (metabolism at rest and standard water loss rate) traits potentially involved in the acclimation response were measured repeatedly during and after water restriction. We observed a sequential acclimation response in water-restricted animals (yearlings spent less time basking during the first weeks of water deprivation) that was followed by delayed sex-specific physiological consequences of the water restriction during the following months (thermal depression in males and lower standard evaporative water loss rates in females). Despite short-term negative effects of water restriction on body growth, annual growth, survival, and reproduction were not significantly different between water-restricted and control yearlings. This demonstrates that beneficial acclimation responses to water restriction involve both short-term flexible behavioral responses and delayed changes in thermal and water biology traits.

How epidemiological patterns shift across populations in an exotic lizard.

Several factors influence the dynamics and structure of parasite communities. Our goal was to investigate how the community composition, prevalence and abundance of parasites change across seven populations of the exotic lizard Hemidactylus mabouia in Northeast Brazil, and to describe ontogenetic and sex variations. We found differences in the composition of component communities and patterns of infection according to the host body size across the lizard populations. We did not find any variation between the sexes regarding epidemiological patterns, which can probably be explained by the similar diet and habitat use of male and female H. mabouia. An unusually high abundance and prevalence of trematodes infecting this host lizard was apparent when we compared other native lizard hosts, and we suggest that local environmental conditions might be advantageous to the development and life cycle of these parasites due to the abundance of all the intermediate and definitive hosts.

Gonadal disruption after single dose exposure of prothioconazole and prothioconazole-desthio in male lizards (Eremias argus).

Prothioconazole (PTC) is a widely used triazole fungicide with low toxicity, and its desulfurization metabolite, prothioconazole-desthio (PTC-d), is reported to have higher reproductive toxicity to mammals. However, little is known about the reproductive toxicity, much less endocrine disrupting effect, of these two chemicals on reptiles. In this study, we investigated the effects of single dose of PTC/PTC-d (100 mg kg-1 body weight) exposure on the pathomorphism of testes and epididymides, serum sex steroid hormones (testosterone and 17ß-estradiol) and transcription of steroidogenic-related genes (STARD, cyp11A, cyp17, cyp19A, 17ß-HSD, 3ß-HSD, AR and ER-α) in gonads of male lizards (Eremias argus). Although structural disorder existed in PTC-d exposure group, severe gonadal disruption, especially suppression of spermatogenesis was only observed in testis after PTC treatment, which consequently led to the lack of spermatozoa in epididymal ducts. Consistent with this result, T/E2 value in PTC exposure was elevated to a significant higher level compared with control and continually increased over time, while T/E2 value in the PTC-d exposure group slightly increased only at 12 h. These results demonstrated a more serious disruption of PTC on male lizard gonads than PTC-d. In addition, the expression of cyp17 gene was inhibited at 6 h, however, was induced at 12 h, and exhibited negative correlations with STARD, cyp11A and 3ß-HSD after PTC exposure at each timepoint. In PTC-d group, the expression of STARD and 3ß-HSD were significantly down-regulated, in contrast, cyp11A and cyp17 were up-regulated, and each gene showed consistent changes over time. For 17ß-HSD, no significance was observed in both treated groups. This study was the first to compare the gonadal disruption of PTC and PTC-d in male lizards and elucidated that these two chemicals influenced the physiological function of male gonads through differential transcriptional modulation.