Landscape metrics as indicators: Quantifying habitat network changes of a bush-cricket Pholidoptera transsylvanica in Hungary

Fragmentation of habitats is a serious problem for many endangered species; a possible solution is the maintenance of landscape connectivity. Due to scarce sources, in managing and planning landscapes exact and quantitative priorities must be set. Application of mathematical tools, such as network analysis, can be useful help in these decisions. We illustrate the possibilities and results of this approach with a case study of endangered Pholidoptera transsylvanica bush-cricket population in the Aggtelek Karst, Northeast-Hungary, which inhabits 39 habitat patches connected with ecological corridors. A key issue in the long-term survival of this metapopulation is the maintenance of gene flow (by preserving the connectivity of the habitat network). We evaluated the landscape graph and our results are compared to earlier ones based on older methods. During the comparison, we used several network indices to set quantitative conservation preferences. In addition, we would like to draw attention to the need for constant monitoring (and possible treatments), because several changes (like secondary succession) have occurred during the years between the two studies, threatening landscape connectivity and long-term survival of certain species. A potential solution for preventing fragmentation is establishing new corridors or improving the existing ones: we estimated the possible effects of these changes. New corridors did not have major effect on the system; maintaining already functioning corridors is more effective.     

Geographic variation in relict populations: genetics and phenotype of bush-cricket Pholidoptera frivaldskyi (Orthoptera) in Carpathians

A decreasing population size is often causing species extinction, however, relict species persisting in small-sized populations counter this. We analysed spatial genetic variation and past changes in population size at the maternally-inherited mitochondrial DNA level to clarify the origin of all recently known isolated populations of Pholidoptera frivaldskyi occurring in the range of Carpathian Mountains. Along with that we analysed also morphological variation as some phenotypic traits can retain useful information on population genetic structure. We found a relatively low genetic diversity within isolated populations as 778 bp COI gene sequences revealed only 13 unique haplotypes (n = 173 individuals from 10 populations). The spatial analysis of molecular variance identified three geographically homogenous genetic clusters (one in Slovakia and two in Romania) with a high level of differentiation among them, suggesting restricted gene flow, whilst Bayesian skyline simulation reconstructed a negative demographic change through evolutionary time. Inferred genetic pattern clearly coincides with differences in males’ colour phenotype as the extent of pigmentation on the lateral pronotum varied significantly among genetic lineages. We suggest that geographical variation in the species populations has relict-like character and their isolated occurrence is not a result of recent introduction events. Identification of ‘evolutionary units’ may help in the conservation and management of this rare insect species.     

Female Bush-Crickets,Pholidoptera griseoaptera,that have Received Smaller Ejaculates Show a Higher Mating Rate in the Field

Females of numerous insect species are known to be polyandrous, but there have been relatively few studies of factors associated with the degree of polyandry in females in the field. Number of copulations by females is negatively associated with ejaculate size across bush-cricket species. Assessing intraspecific variability is important when looking for and interpreting trait evolution. Therefore the aim of this study was to test the association between ejaculate size (i.e. volume of spermatodose–spermatophore-like structure formed within the spermatheca) and mating rate (i.e. number of spermatodoses) of females of Pholidoptera griseoaptera, while accounting for female body size (pronotum length) and age (number of hind leg’s cuticular bands). The results based on field-caught individuals suggested that there were statistically significant negative association between smallest and mean spermatodose volume, respectively, and number of copulations in this nuptial gift-giving bush-cricket species. This is in accordance with interspecific associations between ejaculate size and polyandry. However, lower slope of the intraspecific relationship may suggest lower importance of the ejaculate size in explaining females’ mating rate variability in this nuptial gift-giving species.     

Isolation and characterization of microsatellite loci in the dark bush cricket,Pholidoptera griseoaptera (Tettigoniidae)

Twelve novel polymorphic microsatellite loci are presented for the dark bush cricket, Pholidoptera griseoaptera. All loci are polymorphic, with up to 37 alleles per locus. These microsatellites will be useful tools for studying the influence of landscape structure and land use intensity in agricultural landscapes on genetic diversity within and among populations of P. griseoaptera.     

Repeated sampling detects gene flow in a flightless ground beetle in a fragmented landscape

Secondary clines level down in the course of time if the gene flow is not interrupted. Temporally repeated sampling of populations in a cline allows the investigation not only of its occurrence but also of the estimation of the amount of ongoing gene flow. We reinvestigated an allozyme gradient in Carabus auronitens populations in the Westphalian Lowlands (northwestern Germany) 15 to 20 years after it had originally been recorded. A total of 977 individuals of this flightless woodland species from 29 sample sites were genotyped at the diallelic Est-1 locus in 2005-2006 and compared to former findings, collected in 1985-1994 from the same populations. Both data sets showed clinal variation. Pairwise differences between the samples of both data sets indicated significant decrease in the steepness of the cline during the past 15 to 20 years. The estimated average gene flow per generation is 0.6% of each beetle population. Ongoing gene flow in the flightless ground beetle C. auronitens led to a less pronounced cline despite a stable degree of fragmentation (and connectivity) of the landscape. Migration and gene flow were obviously enabled by the numerous hedgerows. The corridors are seen to be a prerequisite for migration between populations and for possible future range shifts of forest insect species.     

Movement and dispersal patterns in the bush cricket Pholidoptera griseoaptera: the role of developmental stage and sex

Abstract.  1. To persist in a highly fragmented environment, such as the intensively used agricultural landscapes of Europe, species may need to successfully disperse between sub-populations. This paper describes a capture – mark – resight experiment to investigate movement and dispersal patterns of the flightless bush cricket species Pholidoptera griseoaptera .
2. Movement parameters showed little difference between juveniles and imagos as well as between males and females. Both developmental stages and sexes moved considerable distances, were cap\able of crossing habitat boundaries, and showed successful inter-patch dispersal through the agricultural matrix.
3. The results suggest that P. griseoaptera exhibits movement behaviours that compensate for its inability to fly. It was concluded that the good dispersal ability exhibited by animals of both sexes and developmental stage is an important factor allowing P. griseoaptera to persist in fragmented agricultural landscapes.     

Management history determines gene flow in a prominent invader

Invasive plants pose substantial threats to protected areas globally. Although management can limit impacts, spread and reinvasion from neighbouring areas into protected areas are a major and an on‐going problem for land managers. However, identifying the main sources of propagules and the dimensions of invasion pathways is challenging. This study used population genetic markers [inter simple sequence repeats (ISSRs) and amplified fragment length polymorphisms (AFLPs)] to infer the source(s) of re‐colonization and dispersal patterns for a typical invader of riparian and terrestrial habitats (Lantana camara) along the Sabie‐Sand catchment, one of the most important river systems flowing into and across South Africa’s flagship protected area, the Kruger National Park (KNP). Results indicate that populations located along the lower reaches of the Sabie and Sand tributaries harboured substantially higher genetic diversity than those in the upper Sabie catchment. Bayesian assignments indicated that the upper Sabie tributary contributed far fewer propagules than the Sand tributary to the lower Sabie River. Current invasion patterns are due to a combination of a major flood event in 2000 and differences in the degree to which the upstream reaches were managed after the flooding. The major flood of 2000 effectively cleared lantana from the riparian areas. However, whereas on‐going management efforts against riparian species in the KNP have been effective, rendering the upper Sabie relatively clear of lantana, only a small part of the Sand tributary falls under jurisdiction of the KNP and has received consistent management attention. The reinvasion of the lower Sabie in the KNP was therefore almost entirely by propagules from the Sand tributary. The study highlights the important role that molecular tools can play in determining dispersal dynamics and directing invasive species management. For invasive plant species that invade both riparian habitats and landscapes away from rivers in protected areas, such as lantana, management must focus on all major sources of propagules to limit reinvasion.     

Apparent widespread gene flow in the predominantly flightless planthopper Tumidagena minuta

1. To determine whether dispersal biology can predict the pattern of population‐genetic variation among insect populations accurately, allozyme variation was assayed for populations of a saltmarsh planthopper, Tumidagena minuta, in which > 99% of the adults are flightless. 2. The pattern of genetic isolation by distance in T. minuta was compared with that in other insects, to determine whether it was similar to isolation by distance in other sedentary insects. 3. In contrast to predictions, the pattern of isolation by distance in T. minuta was most similar to that seen in the most mobile insects in a recent review of population‐genetic variation in insects. Furthermore, population‐genetic subdivision over a spatial scale of > 400 km was weak. 4. Possible causes of the apparent contradiction between dispersal biology and population‐genetic structure in this species are discussed. The results for T. minuta highlight the fact that although mobility is generally correlated with gene flow in insects, studies of population‐genetic variation must be combined with direct studies of dispersal to understand fully the degree to which populations exchange individuals.     

Snow evens fragmentation effects and food determines overwintering success in ground-dwelling voles

Climate instability strongly affects overwintering conditions in organisms living in a strongly seasonal environment and consequently their survival and population dynamics. Food, predation and density effects are also strong during winter, but the effect of fragmentation of ground vegetation on ground-dwelling small mammals is unknown. Here, we report the results of a winter experiment on the effects of habitat fragmentation and food on experimental overwintering populations of bank voles Myodes glareolus. The study was conducted in large outdoor enclosures containing one large, two medium-sized or four small habitat patches or the total enclosure area covered with protective tall-grass habitat. During the stable snow cover of midwinter, only food affected the overwintering success, body condition, trappability and earlier onset of breeding in bank voles. However, after the snow thaw in spring, habitat fragmentation gained importance again, and breeding activities increased the movements of voles in the most fragmented habitat. The use of an open, risky matrix area increased along the habitat fragmentation. Our experiment showed that long-lasting stable snow cover protects overwintering individuals in otherwise exposed and risky ground habitats. We conclude that a stable winter climate and snow cover should even out habitat fragmentation effects on small mammals. However, along prolonged snow-free early winter and in an earlier spring thaw, this means loss of protection by snow cover both in terms of thermoregulation and predation. Thus, habitat cover is important for the survival of small ground-dwelling boreal mammals also during the non-breeding season.     

Paternity analysis of wild‐caught females shows that sperm package size and placement influence fertilization success in the bushcricket Pholidoptera griseoaptera

In species where females store sperm, males may try to influence paternity by the strategic placement of sperm within the female's sperm storage organ. Sperm may be mixed or layered in storage organs, and this can influence sperm use beyond a ‘fair raffle’. In some insects, sperm from different matings is packaged into discrete packets (spermatodoses), which retain their integrity in the female's sperm storage organ (spermatheca), but little is known about how these may influence patterns of sperm use under natural mating conditions in wild populations. We examined the effect of the size and position of spermatodoses within the spermatheca and number of competing ejaculates on sperm use in female dark bushcrickets (Pholidoptera griseoaptera) that had mated under unmanipulated field conditions. Females were collected near the end of the mating season, and seven hypervariable microsatellite loci were used to assign paternity of eggs laid in the laboratory. Females contained a median of three spermatodoses (range 1–6), and only six of the 36 females contained more than one spermatodose of the same genotype. Both the size and relative placement of the spermatodoses within the spermatheca had a significant effect on paternity, with a bias against smaller spermatodoses and those further from the single entrance/exit of the spermatheca. A higher number of competing males reduced the chances of siring offspring for each male. Hence, both spermatodose size and relative placement in the spermatheca influence paternity success.     

Landscape discontinuities influence gene flow and genetic structure in a large, vagile Australian mammal, Macropus fuliginosus

Large vagile mammals typically exhibit little genetic structuring across their range, particularly when their habitat is essentially continuous. We investigated the population genetic structure of a large vagile Australian macropodid, Macropus fuliginosus, which is continuously distributed across most of southern Australia, using nine highly polymorphic nuclear microsatellite loci. Five distinct genetic units were identified across the range, four on the mainland and one on Kangaroo Island. In addition to the predicted historic Nullarbor Plain Barrier, two unexpected mainland barriers to gene flow were identified. Both were associated with landscape discontinuities (Swan River, Flinders Ranges), which appear within the dispersal capabilities of M. fuliginosus. Typical of large vagile mammals, M. fuliginosus displays high genetic diversity (with the exception of an insular population) and weak genetic structuring (within genetic units). However, the expansion of M. fuliginosus from southwestern Australia during the Pleistocene has resulted in significantly reduced genetic diversity in eastern populations. No significant sex-biased dispersal was detected, although differences in habitat, densities and climatic conditions between the eastern and western regions of the range appear to influence dispersal with the effects of isolation by distance only evident in the west. These results suggest that the biogeography of southern Australia is more complex than previously thought and reveal that seemingly minor landscape features can significantly impact genetic structuring in large vagile mammals.     

Landscape configuration and diversity hotspots in wintering sparrows

Community-level studies with finches have traditionally viewed local resources as the primary constrainst on local diversity. Patches have been considered to be self-contained and embedded in landscapes that were neutral with respect to the ecological processes under investigation. This study uses a factorial design to examine the relative roles of patch content and patch context in determining patterns of species richness. Sparrows were surveyed in small fallow patches that varied in both weed cover and the type of adjacent habitat. Species richness and total sparrow abundance were significantly influenced by both factors. Individual species were also influnced by both factors; however, responses were species-specific. Because occupation of particular plot types was conditional on their association with specific habitat types, the spatial patterning of species assemblages results from the configuration of patch types within the landscape.     

Characteristics of a simple, high-resolution flow cytometer based o a new flow configuration.

A method is described for the quantitative determination of free and bound solute concentrations in the cytoplasm of intact cells. The method includes (a) introduction of a gelatin gel reference phase (RP) into the cytoplasm; (b) diffusion of dissolved substances between cytoplasm and RP, (c) cell quenching to - 196 degrees C to prevent subsequent solute redistributions, (d) ultra-low temperature microdissection to isolate RP and cytoplasm samples, and (e) analysis of isolates for solute and water content. In normal oocytes of the salamander, Desmognathus ochrophaeus, free or RP Na+ and K+ are 21.0 +/- 1.1 and 128.8 +/- 2.4 mu eq/ml, respectively, and vary stoichiometrically in altered oocytes. Overall cytoplasmic concentrations are 75.2 +/- 2.7 mu eq Na+/ml and 88.6 +/- 1.5 mu eq K+/ml. Cytoplasmic chemical activities are 16.2 mu eq Na+/ml and 99.2 mu eq K+/ml, corresponding to activity coefficients of 0.22 and 1.12, respectively. The results demonstrate unambiguously that (a) oocytes actively transport Na+ and K+, and (b) cytoplasm has important binding properties which differentiate it from an ordinary aqueous solution. These cytoplasmic properties are investigated in the following paper.     

Landscape effects on gene flow for a climate‐sensitive montane species,the American pika

Climate change is arguably the greatest challenge to conservation of our time. Most vulnerability assessments rely on past and current species distributions to predict future persistence but ignore species' abilities to disperse through landscapes, which may be particularly important in fragmented habitats and crucial for long‐term persistence in changing environments. Landscape genetic approaches explore the interactions between landscape features and gene flow and can clarify how organisms move among suitable habitats, but have suffered from methodological uncertainties. We used a landscape genetic approach to determine how landscape and climate‐related features influence gene flow for American pikas (Ochotona princeps) in Crater Lake National Park. Pikas are heat intolerant and restricted to cool microclimates; thus, range contractions have been predicted as climate changes. We evaluated the correlation between landscape variables and genetic distance using partial Mantel tests in a causal modelling framework, and used spatially explicit simulations to evaluate methods of model optimization including a novel approach based on relative support and reciprocal causal modelling. We found that gene flow was primarily restricted by topographic relief, water and west‐facing aspects, suggesting that physical restrictions related to small body size and mode of locomotion, as well as exposure to relatively high temperatures, limit pika dispersal in this alpine habitat. Our model optimization successfully identified landscape features influencing resistance in the simulated data for this landscape, but underestimated the magnitude of resistance. This is the first landscape genetic study to address the fundamental question of what limits dispersal and gene flow in the American pika.     

Litter size variation in hypothalamic gene expression determines adult metabolic phenotype in Brandt's voles (Lasiopodomys brandtii)

Background

Early postnatal environments may have long-term and potentially irreversible consequences on hypothalamic neurons involved in energy homeostasis. Litter size is an important life history trait and negatively correlated with milk intake in small mammals, and thus has been regarded as a naturally varying feature of the early developmental environment. Here we investigated the long-term effects of litter size on metabolic phenotype and hypothalamic neuropeptide mRNA expression involved in the regulation of energy homeostasis, using the offspring reared from large (10–12) and small (3–4) litter sizes, of Brandt''s voles (Lasiopodomys brandtii), a rodent species from Inner Mongolia grassland in China.

Methodology/Principal Findings

Hypothalamic leptin signaling and neuropeptides were measured by Real-Time PCR. We showed that offspring reared from small litters were heavier at weaning and also in adulthood than offspring from large litters, accompanied by increased food intake during development. There were no significant differences in serum leptin levels or leptin receptor (OB-Rb) mRNA in the hypothalamus at weaning or in adulthood, however, hypothalamic suppressor of cytokine signaling 3 (SOCS3) mRNA in adulthood increased in small litters compared to that in large litters. As a result, the agouti-related peptide (AgRP) mRNA increased in the offspring from small litters.

Conclusions/Significance

These findings support our hypothesis that natural litter size has a permanent effect on offspring metabolic phenotype and hypothalamic neuropeptide expression, and suggest central leptin resistance and the resultant increase in AgRP expression may be a fundamental mechanism underlying hyperphagia and the increased risk of overweight in pups of small litters. Thus, we conclude that litter size may be an important and central determinant of metabolic fitness in adulthood.     

Initial diapause and embryonic development in the speckled bush-cricket, Leptophyes punctatissima

ABSTRACT. The initiation and pattern of embryonic development in Leptophyes punctatissima (Bosc) (Tettigoniidae) are shown to be variously dependent on temperature. Immediate high temperature (30°C) facilitated rapid and direct embryogenesis to the stage of diapause in the fully formed embryo. Slightly lower temperature (25°C) resulted in a delay before embryogenesis started, and this delay was greatly extended as an initial diapause if incubation temperatures were decreased (20-16°C). Still lower initial temperatures (8–12°C) facilitated subsequent development at 20–30°C. These responses all increased as a function of exposure time. Once initiated, the rate of development was temperature dependent, but competence to tolerate high temperature, and ability to continue development at low temperatures, changed with age. In general, the developmental temperature range appears to be lowered with age. None of these different treatments had any effect on the late embryonic diapause. According to the temperatures prevailing at oviposition, Leptophyes could be an annual or a biennial species; the biennial pattern is normal.     

Sulfoxide configuration in sparsomycin determines time-dependent and competitive inhibition of peptidyl transferase

Sparsomycin, S_cR_s configuration, was the most potent of the four possible stereoisomers as a competitive inhibitor of peptide bond formation. In addition, the configuration of the two chiral centers dictated whether the compound exhibited time- and temperature-dependent inhibition of peptidyl transferase when incubated with polysomes prior to enzyme assay. The data corroborate the thesis that a peptidyl transferase-mediated acylation of the pivotal sulfoxide moiety and subsequent Pummerer rearrangement play a significant role in the inhibitory properties of sparsomycin.     

Post-eclosion diuresis in a flightless insect, the silkmoth Bombyx mori

Abstract. Although the silkmoth, Bombyx mori L., has lost the ability to fly, it has retained a post-eclosion diuresis. In moths removed from their cocoons before eclosion, or in those which failed to spin cocoons as larvae, the weight loss due to diuresis was 14% of the eclosion body weight in males. Moths which used labial fluid to escape from their cocoons showed a correspondingly smaller diuresis (5%). Both urine and labial fluid had high potassium and low sodium concentrations. Unlike post-eclosion diuresis in butterflies, however, the urine was isosmotic to the haemolymph. In vitro preparations of B.mori Malpighian tubules were stimulated by cyclic AMP, B.mori brain extracts and Manduca sexta diuretic peptide (Mas-DP I).     

Landscape heterogeneity predicts gene flow in a widespread polymorphic bumble bee, Bombus bifarius (Hymenoptera: Apidae)

Bombus bifarius is a widespread bumble bee that occurs in montane regions of western North America. This species has several major color pattern polymorphisms and shows evidence of genetic structuring among regional populations, and the taxonomic status of regional populations has repeatedly been debated. We test whether observed structure is evidence for discrete gene flow barriers that might indicate isolation or instead reflects clinal variation associated with spatially limited dispersal in a complex landscape. We first consider color pattern variation and identify geographical patterns of B. bifarius color variation using cluster analysis. We then use climate data and a comprehensive set of B. bifarius natural history records with an existing genetic data set to model the distribution of environmentally suitable habitat in western North America and predict pathways of potential gene flow using circuit theory. Resistance distances among populations that incorporate environmental suitability information predict patterns of genetic structure much better than geographic distance or Bayesian clustering alone. Results suggest that there may not be barriers to gene flow warranting further taxonomic considerations, but rather that the arrangement of suitable habitat at broad scales limits dispersal sufficiently to explain observed levels of population differentiation in B. bifarius.