Phenotypic plasticity and local adaptations to dissolved oxygen in larvae fire salamander (Salamandra infraimmaculata).

A key environmental factor that varies both spatially and temporally in surface waters is dissolved oxygen (DO). In stagnant ephemeral freshwater ponds, DO can fluctuate diurnally and seasonally, while the constant mixing of water in streams typically maintain DO levels close to saturation with only minor fluctuations. Larvae of the Near Eastern fire salamander (Salamandra infraimmaculata) develop in a range of waterbodies that vary in flow and permanence. To study inter-population variation in larval response to environmental change, we translocated larvae between stream and pond habitats and exposed larvae sampled from different habitat types to hypoxic and normoxic conditions in the laboratory. Larvae transferred from stream to pond retain gill size, while larvae transferred from pond to stream show a reduction in gill size. Larvae that were caged within their native habitat, either stream or pond, display a decrease in gill size similar to larvae transferred from pond to stream. When exposed to experimentally manipulated levels of DO in the laboratory larvae, respectively, increase and decrease gill size under hypoxic and normoxic conditions. Habitat-type origin had a significant effect on the degree of change in gill size with larvae from permanent streams demonstrating the lowest absolute variation in gill size. There was no interaction between DO level (hypoxic/normoxic) and the larvae habitat-type origin. These results suggest that S. infraimmaculata larvae are locally adapted to their aquatic breeding habitat through the plastic ability to respond to the prevailing respiratory conditions by rapidly decreasing or increasing gill size.

Green roof and photovoltaic panel integration: Effects on plant and arthropod diversity and electricity production.

The combination of green roofs with photovoltaic (PV) panels has been proposed to provide synergistic benefits as the panel is cooled by the presence of the vegetation, and thus produces more electricity, while the solar panel enhances growing conditions for vegetation, and increases abiotic heterogeneity, resulting in higher plant diversity. We tested these hypotheses in a non-irrigated green roof in a Mediterranean climate with replicated plots including green roofs only, green roofs with a PV panel, and a conventional roof surface with a PV panel. We found that presence of a panel resulted in higher heterogeneity in substrate moisture, but there was no effect on plant diversity. Plant species showed enhanced growth in plots with PV, including greater growth of Sedum sediforme and longer flowering time of annual species. On the other hand, arthropod diversity was lower during part of the year, and abundance of some arthropod taxa was lower in green roof plots with PV. The presence of the green roof also did not improve electricity production by the panels. We conclude that in a Mediterranean climate, it would be appropriate to examine the use of irrigation in green roofs with PV panels, including effects on the plant community and on electricity production.

Inferring the shallow phylogeny of true salamanders (Salamandra) by multiple phylogenomic approaches.

The rise of high-throughput sequencing techniques provides the unprecedented opportunity to analyse controversial phylogenetic relationships in great depth, but also introduces a risk of being misinterpreted by high node support values influenced by unevenly distributed missing data or unrealistic model assumptions. Here, we use three largely independent phylogenomic data sets to reconstruct the controversial phylogeny of true salamanders of the genus Salamandra, a group of amphibians providing an intriguing model to study the evolution of aposematism and viviparity. For all six species of the genus Salamandra, and two outgroup species from its sister genus Lyciasalamandra, we used RNA sequencing (RNAseq) and restriction site associated DNA sequencing (RADseq) to obtain data for: (1) 3070 nuclear protein-coding genes from RNAseq; (2) 7440 loci obtained by RADseq; and (3) full mitochondrial genomes. The RNAseq and RADseq data sets retrieved fully congruent topologies when each of them was analyzed in a concatenation approach, with high support for: (1) S. infraimmaculata being sister group to all other Salamandra species; (2) S. algira being sister to S. salamandra; (3) these two species being the sister group to a clade containing S. atra, S. corsica and S. lanzai; and (4) the alpine species S. atra and S. lanzai being sister taxa. The phylogeny inferred from the mitochondrial genome sequences differed from these results, most notably by strongly supporting a clade containing S. atra and S. corsica as sister taxa. A different placement of S. corsica was also retrieved when analysing the RNAseq and RADseq data under species tree approaches. Closer examination of gene trees derived from RNAseq revealed that only a low number of them supported each of the alternative placements of S. atra. Furthermore, gene jackknife support for the S. atra - S. lanzai node stabilized only with very large concatenated data sets. The phylogeny of true salamanders thus provides a compelling example of how classical node support metrics such as bootstrap and Bayesian posterior probability can provide high confidence values in a phylogenomic topology even if the phylogenetic signal for some nodes is spurious, highlighting the importance of complementary approaches such as gene jackknifing. Yet, the general congruence among the topologies recovered from the RNAseq and RADseq data sets increases our confidence in the results, and validates the use of phylotranscriptomic approaches for reconstructing shallow relationships among closely related taxa. We hypothesize that the evolution of Salamandra has been characterized by episodes of introgressive hybridization, which would explain the difficulties of fully reconstructing their evolutionary relationships.

Intercohort size structure dynamics of fire salamander larvae in ephemeral habitats: a mesocosm experiment.

The size structure of a larval population facilitates interaction asymmetries that, in turn, influence the dynamics of size-structure. In species that exhibit conspicuous aggressive interactions, the competitive effects of the smaller individuals may be overlooked. We manipulated initial size differences between two larval cohorts and young-cohort density of Salamandra infraimmaculata in mesocosms to determine: (1) whether young individuals function primarily as prey or as competitors of older and larger individuals; (2) the resulting dynamics of size variation; and (3) recruitment to the postmetamorph population. Intercohort size differences generally remained constant over time at low young-cohort densities, but reduced over time at high densities due to retardation of the old-cohort growth rate. This suggests a competitive advantage to the young cohort that outweighs the interference advantage of older cohorts previously documented in this species. The increase in mortality from desiccation due to high young-cohort density was an order of magnitude greater in the old cohort than in the young-cohort, further indicating size-dependent vulnerability to competition. However, the conditions least favorable to most of the old-cohort larvae (large size difference and high young-cohort density) promoted cannibalism. Among cannibals, mortality and time to metamorphosis decreased and sizes at metamorphosis increased substantially. Thus, a balance between the competitive advantage to young cohorts, and the interference and cannibalism advantage to old cohorts shapes larval size-structure dynamics. Larval densities and individual expression of cannibalism can shift this balance in opposite directions and alter relative recruitment rates from different cohorts.

Interactive effects of salinity and a predator on mosquito oviposition and larval performance.

Oviposition habitat selection (OHS) is increasingly being recognized as playing a large role in explaining mosquito distributions and community assemblages. Most studies have assessed the role of single factors affecting OHS, while in nature, oviposition patterns are most likely explained by multiple, interacting biotic and abiotic factors. Determining how various factors interact to affect OHS is important for understanding metapopulation and metacommunity dynamics. We investigated the individual and interactive effects of three water salinities (0, 15 and 30 p.p.t. NaCl added) and the aquatic predator Anisops debilis Perplexa (Hemiptera: Notonectidae) on OHS and larval performance of the mosquitoes Ochlerotatus caspius Pallas and Culiseta longiareolata Macquart (Diptera: Culicidae) in outdoor-artificial-pool and laboratory experiments. C. longiareolata inhabited only freshwater pools, strongly avoided pools containing A. debilis, and larvae experienced lower survival in the presence of A. debilis. Salinity concentration interacted strongly with the predator in affecting OHS and larval survival of O. caspius; oviposition increased with increasing salinity in the absence of the predator and decreased with increasing salinity in the presence of the predator. O. caspius larval survival in predator-free pools was lowest in freshwater and highest at intermediate salinity. In predator pools, survival was highest at high salinity, where predation rate was shown to be lowest in the laboratory. Our results highlight that assessing the role of single factors in affecting mosquito distributions can be misleading. Instead, multiple factors may interact to affect oviposition patterns and larval performance.

Landscape influences on dispersal behaviour: a theoretical model and empirical test using the fire salamander, Salamandra infraimmaculata.

When populations reside within a heterogeneous landscape, isolation by distance may not be a good predictor of genetic divergence if dispersal behaviour and therefore gene flow depend on landscape features. Commonly used approaches linking landscape features to gene flow include the least cost path (LCP), random walk (RW), and isolation by resistance (IBR) models. However, none of these models is likely to be the most appropriate for all species and in all environments. We compared the performance of LCP, RW and IBR models of dispersal with the aid of simulations conducted on artificially generated landscapes. We also applied each model to empirical data on the landscape genetics of the endangered fire salamander, Salamandra infraimmaculata, in northern Israel, where conservation planning requires an understanding of the dispersal corridors. Our simulations demonstrate that wide dispersal corridors of the low-cost environment facilitate dispersal in the IBR model, but inhibit dispersal in the RW model. In our empirical study, IBR explained the genetic divergence better than the LCP and RW models (partial Mantel correlation 0.413 for IBR, compared to 0.212 for LCP, and 0.340 for RW). Overall dispersal cost in salamanders was also well predicted by landscape feature slope steepness (76%), and elevation (24%). We conclude that fire salamander dispersal is well characterised by IBR predictions. Together with our simulation findings, these results indicate that wide dispersal corridors facilitate, rather than hinder, salamander dispersal. Comparison of genetic data to dispersal model outputs can be a useful technique in inferring dispersal behaviour from population genetic data.

Syntactic structure and geographical dialects in the songs of male rock hyraxes.

Few mammalian species produce vocalizations that are as richly structured as bird songs, and this greatly restricts the capacity for information transfer. Syntactically complex mammalian vocalizations have been previously studied only in primates, cetaceans and bats. We provide evidence of complex syntactic vocalizations in a small social mammal: the rock hyrax (Procavia capensis: Hyracoidea). We adopted three algorithms, commonly used in genetic sequence analysis and information theory, to examine the order of syllables in hyrax calls. Syntactic dialects exist, and the syntax of hyrax calls is significantly different between different regions in Israel. Call syntax difference is positively correlated to geographical distance over short distances. No correlation is found over long distances, which may reflect limited dispersal movement. These findings indicate that rich syntactic structure is more common in the vocalizations of mammalian taxa than previously thought and suggest the possibility of vocal production learning in the hyrax.

Predator-released hydrocarbons repel oviposition by a mosquito.

Prey species commonly use predator-released kairomones (PRKs) to detect risk of predation, yet the chemical identity of PRKs remains elusive. Chemical identification of PRKs will facilitate the study of predator-prey interactions and the risk of predation, and when the prey are pests, will potentially provide environmentally friendly means of pest control. In temporary pools of the Mediterranean and Middle East, larvae of the mosquito Culiseta longiareolata Macquart are highly vulnerable to the common predatory backswimmer, Notonecta maculata Fabricius. We demonstrate that N. maculata releases two hydrocarbons, n-heneicosane and n-tricosane, which repel ovipositing females of C. longiareolata. In behavioural tests with environmentally relevant chemical concentrations in outdoor mesocosm experiments, the repellent effects of the two compounds were additive at the tested concentrations.

Context-dependent reproductive habitat selection: the interactive roles of structural complexity and cannibalistic conspecifics.

Structural complexity generally reduces predation and cannibalism rates. Although the benefits from this effect vary among environmental contexts and through time, it has been the common explanation for high species abundance in complex habitats. We hypothesized that oviposition habitat selection for structural complexity depends on the expected trophic function of the progeny. In Salamandra infraimmaculata larvae, expected trophic function is dictated by their sequence of deposition. First cohorts cannibalize later-arriving cohorts, while all compete for shared prey resources. In a mesocosm experiment, we show that gravid salamanders facing conspecific-free pools preferred structurally simple habitats (no rocks), while females facing only pools with older conspecific larvae preferred complex habitats (with rocks). Context-dependent preference of habitat complexity for managing food/safety trade-offs may be extended from classic foraging patch decisions to breeding habitat selection. These trade-offs vary with dynamic larval processes such as priority effects and ontogenetic diet shifts, potentially leading to complex maternal parturition behaviours.

Fine-scale substrate heterogeneity in green roof plant communities: The constraint of size.

Heterogeneity-diversity relationship (HDR) is commonly shown to be positive in accordance with classic niche processes. However, recent soil-based studies have often found neutral and even negative HDRs. Some of the suggested reasons for this discrepancy include the lack of resemblance between manipulated substrate and natural settings, the treated areas not being large enough to contain species' root span, and finally limited-sized plots may not sustain focal species' populations over time. Vegetated green roofs are a growing phenomenon in many cities that could be an ideal testing ground for this problem. Recent studies have focused on the ability of these roofs to sustain stable and diverse plant communities and substrate heterogeneity that would increase niches on the roof has been proposed as a method to attain this goal. We constructed an experimental design using green roof experimental modules (4 m2) where we manipulated mineral and organic substrate component heterogeneity in different subplots (0.25 m2) within the experimental module while maintaining the total sum of mineral and organic components. A local annual plant community was seeded in the modules and monitored over three growing seasons. We found that plant diversity and biomass were not affected by experimentally created substrate heterogeneity. In addition, we found that different treatments, as well as specific subplot substrates, had an effect on plant community assemblages during the first year but not during the second and third years. Substrate heterogeneity levels were mostly unchanged over time. The inability to retain plant community composition over the years despite the maintenance of substrate differences supports the hypothesis that maintenance of diversity is constrained at these spatial scales by unfavorable dispersal and increased stochastic events as opposed to predictions of classic niche processes.

Fine-scale substrate heterogeneity does not affect arthropod communities on green roofs.

Green roofs, which are roofs with growing substrate and vegetation, can provide habitat for arthropods in cities. Maintaining a diversity of arthropods in an urban environment can enhance the functions they fill, such as pest control and soil development. Theory suggests that the creation of a heterogeneous environment on green roofs would enhance arthropod diversity. Several studies have examined how arthropod diversity can be enhanced on green roofs, and particularly whether substrate properties affect the arthropod community, but a gap remains in identifying the effect of substrate heterogeneity within a green roof on the arthropod community. In this paper, it is hypothesized that creating heterogeneity in the substrate would directly affect the diversity and abundance of some arthropod taxa, and indirectly increase arthropod diversity through increased plant diversity. These hypotheses were tested using green roof plots in four treatments of substrate heterogeneity: (1) homogeneous dispersion; (2) mineral heterogeneity-with increased tuff concentration in subplots; (3) organic heterogeneity-with decreased compost concentrations in subplots; (4) both mineral and organic heterogeneity. Each of the four treatments was replicated twice on each of three roofs (six replicates per treatment) in a Mediterranean region. There was no effect of substrate heterogeneity on arthropod diversity, abundance, or community composition, but there were differences in arthropod communities among roofs. This suggests that the location of a green roof, which can differ in local climatic conditions, can have a strong effect on the composition of the arthropod community. Thus, arthropod diversity may be promoted by building green roofs in a variety of locations throughout a city, even if the roof construction is similar on all roofs.

Comparing grey water versus tap water and coal ash versus perlite on growth of two plant species on green roofs.

Green roofs provide important ecosystem services in urban areas. In Mediterranean and other semi-arid climate regions, most perennial plants on green roofs need to be irrigated during the dry season. However, the use of freshwater in such regions is scarce. Therefore, the possibility of using grey water should be examined. Coal ash, produced primarily from the burning of coal in power plants, constitutes an environmental contaminant that should be disposed. One option is to use ash as a growing substrate for plants. Here, we compare the effects of irrigating with grey- versus tap-water and using ash versus perlite as growing substrates in green roofs. The study was conducted in northern Israel in a Mediterranean climate. The design was full factorial with three factors: water-type (grey or tap-water)×substrate-type (coal ash vs perlite)×plant species (Phyla nodiflora, Convolvulus mauritanicus or no-plant). The development of plants and the quality of drainage water along the season, as well as quality of the used substrates were monitored. Both plant species developed well under all the experimental conditions with no effect of water type or substrate type. Under all treatments, both plant species enhanced electrical conductivity (EC) and chemical oxygen demand (COD) of the drainage water. In the summer, EC and COD reached levels that are unacceptable in water and are intended to be reused for irrigation. We conclude that irrigating with grey water and using coal ash as a growth substrate can both be implemented in green roofs. The drainage from tap water as well as from grey water can be further used for irrigating the roof, but for that, COD and EC levels must be lowered by adding a sufficient amount of tap water before reusing.

Sedum-dominated green-roofs in a semi-arid region increase CO2 concentrations during the dry season.

Green roofs are expected to absorb and store carbon in plants and soils and thereby reduce the high CO2 concentration levels in big cities. Sedum species, which are succulent perennials, are commonly used in extensive green roofs due to their shallow root system and ability to withstand long water deficiencies. Here we examined CO2 fixation and emission rates for Mediterranean Sedum sediforme on green-roof experimental plots. During late winter to early spring, we monitored CO2 concentrations inside transparent tents placed over 1m2 plots and followed gas exchange at the leaf level using a portable gas-exchange system. We found high rates of CO2 emission at daytime, which is when CO2 concentration in the city is the highest. Both plot- and leaf-scale measurements showed that these CO2 emissions were not fully compensated by the nighttime uptake. We conclude that although carbon sequestration may only be a secondary benefit of green roofs, for improving this ecosystem service, other plant species than Sedum should also be considered for use in green roofs, especially in Mediterranean and other semi-arid climates.

Effects of pool depth and risk of predation on oviposition habitat selection by temporary pool dipterans.

Natural selection should favor females that oviposit in sites providing the best available conditions for her progeny. In temporary pools, two important conditions include risk of desiccation and risk of predation to larvae. In an artificial pool experiment, we compared oviposition responses of two dipterans, Culiseta longiareolata Macquart (Culicidae) and Chironomus riparius Meigan (Chironomidae), in very shallow pools that, if left to dry, would not have had a long enough hydroperiod for larvae to complete development, versus deeper pools that would have a sufficiently long enough hydroperiod for development. Water depth was crossed with the presence or absence of the predatory backswimmer Notonecta maculata F. (Hemiptera: Notonectidae). Oviposition patterns were consistent with larval vulnerability of the two species to predation by N. maculata. C. longiareolata, whose larvae are highly vulnerable to predation, strongly avoided ovipositing in pools containing this predator, whereas C. riparius, whose larvae are considerably less vulnerable, did not display oviposition avoidance. Pool depth did not affect oviposition habitat selection in either dipteran.

Chemical detection of the predator Notonecta irrorata by ovipositing Culex mosquitoes.

We tested the oviposition response of Culex mosquitoes to the predator Notonecta irrorata in an outdoor artificial pool experiment employing equal numbers of control and predator pools. There was a strong oviposition avoidance by Culex of Notonecta pools; 83% of egg rafts were found in control pools during the period in which Notonecta were present. After removing Notonecta, mosquitoes continued to avoid ovipositing in the former Notonecta pools for two additional days suggesting a predator-released kairomone as the cue used by the mosquitoes to detect the presence of this predator.

The taxonomic status of Dugesiabiblica from Israel and Turkey (Platyhelminthes, Tricladida, Dugesiidae).

The taxonomic status of Dugesiabiblica (Platyhelminthes, Tricladida, Dugesiidae) from Israel and Turkey is problematic due to its morphological similarity with Dugesiasicula since these nominal species present overlapping characters. In this study we analyzed histological preparations of specimens of these two nominal species and also compared mitochondrial COI gene sequences from Israeli populations to the already known haplotype composition of Dugesiasicula. We concluded that these animals belong to the same species and therefore we consider Dugesiabiblica to be a junior synonym of Dugesiasicula. This implies that the distribution range of Dugesiasicula is even wider than previously thought, and that the species is present all around the Mediterranean Basin and on many of its islands.

Effects of Tail Clipping on Larval Performance and Tail Regeneration Rates in the Near Eastern Fire Salamander, Salamandra infraimmaculata.

Tail-tip clipping is a common technique for collecting tissue samples from amphibian larvae and adults. Surprisingly, studies of this invasive sampling procedure or of natural tail clipping--i.e., bites inflicted by predators including conspecifics--on the performance and fitness of aquatic larval stages of urodeles are scarce. We conducted two studies in which we assessed the effects of posterior tail clipping (~30 percent of tail) on Near Eastern fire salamander (Salamandra infraimmaculata) larvae. In a laboratory study, we checked regeneration rates of posterior tail-tip clipping at different ages. Regeneration rates were hump-shaped, peaking at the age of ~30 days and then decreasing. This variation in tail regeneration rates suggests tradeoffs in resource allocation between regeneration and somatic growth during early and advanced development. In an outdoor artificial pond experiment, under constant larval densities, we assessed how tail clipping of newborn larvae affects survival to, time to, and size at metamorphosis. Repeated measures ANOVA on mean larval survival per pond revealed no effect of tail clipping. Tail clipping had correspondingly no effect on larval growth and development expressed in size (mass and snout-vent length) at, and time to, metamorphosis. We conclude that despite the given variation in tail regeneration rates throughout larval ontogeny, clipping of 30% percent of the posterior tail area seems to have no adverse effects on larval fitness and survival. We suggest that future use of this imperative tool for the study of amphibian should take into account larval developmental stage during the time of application and not just the relative size of the clipped tail sample.

Non-consumptive effects of larval Salamandra on crustacean prey: can eggs detect predators?

Predators affect prey populations not only by prey consumption but also in nonconsumptive ways including modifying prey behavior. I tested the effects of fire salamander larvae (Salamandra infraimmaculata) on populations of co-occurring crustacean species in artificial outdoor pools. I also tested whether these effects were due entirely to prey consumption by Salamandra larvae or alternatively to some nonconsumptive effect. The soil (containing crustacean eggs) added to the artificial pools was collected from a dried-out temporary pool that is inhabited by Salamandra during the early part of the hydroperiod. I randomly assigned the pools to one of three treatments: control, free Salamandra, or caged Salamandra. Free salamander larvae could roam the entire pool and prey upon crustaceans. Caged salamander larvae were placed within a cage with having 250-µm mesh windows. They could not prey upon the crustaceans but could, for example, influence them by chemical cues. Densities of the three dominant crustacean species (Arctodiaptomus similis, Ceriodaphnia quadrangula and Cyzicus sp.) were drastically reduced in both salamander treatments compared to the control. Crustacean densities, however, were not significantly different between the two salamander treatments. One plausible explanation is that crustacean eggs can detect the presence of this predator via chemical cues and delay hatching.

Inter- and intra-specific density-dependent effects on life history and development strategies of larval mosquitoes.

We explored how inter- and intra-specific competition among larvae of two temporary-pool mosquito species, Culiseta longiareolata and Ochlerotatus caspius, affect larval developmental strategy and life history traits. Given that their larvae have similar feeding habits, we expected negative reciprocal inter-specific interactions. In a microcosm experiment, we found sex-specific responses of larval survival and development to both intra- and inter-specific larval competition. C. longiareolata was the superior competitor, reducing adult size and modifying larval developmental time of O. caspius. We observed two distinct waves of adult emergence in O. caspius, with clear sex-specific responses to its inter-specific competitor. In males, this pattern was not affected by C. longiareolata, but in females, the timing and average body size of the second wave strongly varied with C. longiareolata density. Specifically, in the absence of C. longiareolata, the second wave immediately followed the first wave. However, as C. longiareolata abundance increased, the second wave was progressively delayed and the resulting females tended to be larger. This study improves our understanding of the way intra- and inter-specific competition combine to influence the life histories of species making up temporary pond communities. It also provides strong evidence that not all individuals of a cohort employ the same strategies in response to competition.