Genetic population structure of two cryptic Gammarus fossarum types across a contact zone

Previous studies have revealed inconsistent results about the taxonomic status of European Gammarus fossarum forms. The variability in morphology and hybridization ability has not shown clear geographic patterns, whereas on a genetical basis two Central European G. fossarum forms have been proposed. In the present study the genetic structure of G. fossarum populations was investigated across a natural contact zone. For the first time direct comparisons of allotopic versus syntopic populations were feasible. 24 Populations of G. fossarum plus 4 populations of G. pulex (as outgroup) were sampled along 2 transects across the contact zone. The genotypes of about 60 individuals per population were revealed by allozyme electrophoresis at 6 polymorphic loci. Parallel step ‘clines’ in allele frequencies indicate secondary contact of the two G. fossarum forms. The level of genetic variation between the forms was comparable to an interspecific level (G. pulex versus G. fossarum). The pronounced genetic bipartition in syntopic G. fossarum populations and the lack of potential F1-hybrids further corroborated the taxonomic status of the G. fossarum types. The geographic patterns of the genetic differentiation between and within the G. fossarum types, the allelic diversities and the genotypic disequilibria revealed recent introgression at the analysed loci to be insignificant. In addition it is shown that the population structure of the western G. fossarum type is out of equilibrium, which suggest a recent colonization of this type probably by displacement of the other type.     

Genetic Diversity of Daphnia pulex in the Middle and Lower Reaches of the Yangtze River

Increased human activities and environmental changes may lead to genetic diversity variations of Cladocerans in water. Daphnia pulex are distributed throughout the world and often regarded as a model organism. The 16S rDNA, cytochrome c oxidase subunit I (COI), and 18S genes were used as molecular marks. The genetic diversity and phylogeny of D. pulex obtained from 10 water bodies in the middle and lower reaches of the Yangtze River were studied. For 16S rDNA, COI gene, and 18S gene, the A+T content (65.4%, 58.4%, and 54.6%) was significantly higher than the G+C content (34.6%, 41.6% and 45.4%). This result was consistent with higher A and T contents among invertebrates. Based on the genetic distances of 16S rDNA and COI genes, the genetic differences of D. pulex from 10 water bodies located in the middle and lower reaches of the Yangtze River in China was minimal (0%–0.8% for 16S rDNA and 0%–1.5% for COI gene). However, D. pulex evolved into two branches in the phylogenetic trees, which coincided with its geographical distribution. Compared with D. pulex from other countries, the average genetic distance of D. pulex obtained from 10 water bodies in the middle and lower reaches of the Yangtze River reached 9.1%–10.5%, thereby indicating that D. pulex may have evolved into different subspecies.     

Effects of temperature, age and body size on moulting and growth in the freshwater amphipods Gammarus fossarum and G. roeseli

1. The total body length, cephalic length, wet weight and dry weight was measured in juveniles, males and females of Gammarus fossarum and G. roeseli kept in the laboratory. Numbers of flagellar segments on the first and second pairs of antennae of G. fossarum and G. roeseli were quantitatively related to body size and in stars. The addition of segments cannot be used to identify particular instars of individuals or to determine their ages in natural populations. 2. At experimental temperatures ranging from 3.8 to 20.2°C, the number of moults, duration of intermoults, maturation times and specific growth rates were studied from birth in isolated specimens. Sexual maturity was reached after 9 or 10 moults, at a mean wet weight of c. 5mg for females and c. 7mg for males of G. fossarum, and at c. 10 mg for females and c. 13 mg for males of G. roeseli. At 3.8°C neither species reached sexual maturity within 550 days. 3. The mean interval between moults was observed from birth to sexual maturity and was linearly related to moult number and exponentially related to age. The relationship between each intermoult interval, or the maturation time, and the experimental water temperature was described by a power function. Maturation times increased from 96 days at 20.2°C to 355 days at 7.9°C in G. fossarum, and 85 days to 403 days in G. roeseli. 4. Over the range 3.8–20.2°C there was a log—log relationship between temperature and specific growth rates. Growth was maximal at 20.2°C in newborn animals as well as in small sexually mature animals; interspecific differences were highly significant. 5. Increase in body wet weight of G. fossarum and G. roeseli fed ad libitum on a constant mixture of autumn-shed, naturally decaying, tree leaves and aquatic macrophytes was followed to senescence and death. The instantaneous or specific growth rate was maximal near birth, at c. 7.98% wet weight day^?1 in G. fossarum and 9.03% in G. roeseli. At ≥12°C, growth conformed to a logistic curve; maximum absolute increments in weight occurred about half-way through a life span of 280–300 days at 20°C, 380–420 days at 16°C and 550–600 days at 12°C. Some individuals lived longer than 850 days at ≤12°C. The wet weight at birth was 0.112mg for G. fossarum and 0.123mg for G. roeseli. Asymptotic mean body weights of males and females were, respectively, 61 and 41 mg for G. fossarum and 87 and 58 mg for G. roeseli. However, G. roeseli reached the inflection point of the logistic curve significantly faster than G. fossarum. In the latter species, growth and maturation were relatively faster at temperatures below 12°C, whereas they were faster in G. roeseli at 16–20°C. Thus G. fossarum is adapted to summer-cool streams and G. roeseli is adapted to summer-warm streams.     

Substantial differences in genetic diversity and spatial structuring among (cryptic) amphipod species in a mountainous river basin

1. Recent advances in molecular methods foster the documentation of small spatial scale biological diversity over large geographical areas. These advances allow to correctly record α-diversity, but also enable biomonitoring that describes intraspecific molecular diversity, providing valuable insights into the contemporary history of species. Such information is essential for the accurate monitoring of freshwater communities and provides a promising tool to identify conservation priorities at various spatial scales.
2. Here, we combined morphological species determinations with genetic characterisation via DNA barcoding and species distribution modelling. We aimed to investigate whether closely related amphipod species occupying overlapping ecological niches and occurring in partial sympatry, demonstrate similar spatial patterns of intraspecific genetic diversity and share comparable population histories. Therefore, we characterised the amphipod fauna within the Kinzig catchment (1,058 km², Hesse, Central Germany) that is a tributary of the Main River and part of the long-term ecological research network using genetics.
3. Our genetic analysis revealed two more taxonomic entities than previously known. The most common species was Gammarus fossarum clade 11 (or type B), followed by Gammarus roeselii clade C, Gammarus pulex clade D, G. pulex clade B and a very rare previously unknown lineage within the G. fossarum-species complex, which we refer to as G. fossarum clade RMO. These five taxa differed in their intraspecific genetic diversity, with G. fossarum clade 11 demonstrating the highest diversity and having a prominent small-scale pattern with endemic haplotypes in headwater regions. Distributions were predicted for the three most abundant molecularly identified species.
4. The upstream reaches maintained high intraspecific α- and β-diversity, pointing towards a more complex population structure of G. fossarum clade 11. This highlights the importance of considering intraspecific diversity for the conservation of individual species. DNA-based species distribution models shed light on species-specific habitat preferences, and showed spatial distribution patterns that supported ecological inference and conservation management. Barcoding specimens prior to modelling can increase robustness and performance of distribution models as juveniles can be incorporated, and cryptic species complexes disentangled.
5. Our integrative study contributes to the further development of science-informed and holistically considered effective conservation measures. Some poorly dispersing hololimnic species may serve as representatives for our understanding of the natural history of the local communities in headwater regions—and their protection. Intraspecific genetic diversity should be considered in conservation management decisions as it can provide valuable information on past and present population demography, connectivity, and recovery processes of species—information that rarely can be achieved by traditional monitoring approaches.

     

Conspecific versus heterospecific gene exchange between populations of Darwin's finches

This study addresses the extent and consequences of gene exchange between populations of Darwin''s finches. Four species of ground finches (Geospiza) inhabit the small island of Daphne Major in the centre of the Galápagos archipelago. We undertook a study of microsatellite DNA variation at 16 loci in order to quantify gene flow within species owing to immigration and between species owing to hybridization. A combination of pedigrees of observed breeders and assignments of individuals to populations by the program Structure enabled us to determine the frequency of gene exchange and the island of origin of immigrants in some cases. The relatively large populations of Geospiza fortis and G. scandens receive conspecific immigrants at a rate of less than one per generation. They exchange genes more frequently by rare but repeated hybridization. Effects of heterospecific gene flow from hybridization are not counteracted by lower fitness of the offspring. As a result, the standing genetic variation of the two main resident populations on Daphne Major is enhanced to a greater extent by introgressive hybridization than through breeding with conspecific immigrants. Immigrant G. fuliginosa also breeds with G. fortis. Conspecific immigration was highest in the fourth species, G. magnirostris. This species is much larger than the other three and perhaps for this reason it has not bred with any of them. The source island of most immigrants is probably the neighbouring island of Santa Cruz. Evolutionary change may be inhibited in G. magnirostris by continuing gene flow, but enhanced in G. fortis and G. scandens by introgressive hybridization.     

Confrontation of cryptic diversity and mate discrimination within Gammarus pulex and Gammarus fossarum species complexes

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How river structure and biological traits influence gene flow: a population genetic study of two stream invertebrates with differing dispersal abilities

1. Determined by landscape structure as well as dispersal‐related traits of species, connectivity influences various key aspects of population biology, ranging from population persistence to genetic structure and diversity. Here, we investigated differences in small‐scale connectivity in terms of gene flow between populations of two ecologically important invertebrates with contrasting dispersal‐related traits: an amphipod (Gammarus fossarum) with a purely aquatic life cycle and a mayfly (Baetis rhodani) with a terrestrial adult stage. 2. We used highly polymorphic markers to estimate genetic differentiation between populations of both species within a Swiss pre‐alpine catchment and compared these results to the broader‐scale genetic structure within the Rhine drainage. Landscape genetic approaches were used to test for correlations of genetic and geographical structures and in‐stream barrier effects. 3. We found overall very weak genetic structure in populations of B. rhodani. In contrast, G. fossarum showed strong genetic differentiation, even at spatial scales of a few kilometres, and a clear pattern of isolation by distance. Genetic diversity decreased from downstream towards upstream populations of G. fossarum, suggesting asymmetric gene flow. Correlation of genetic structure with landscape topography was more pronounced in the amphipod. Our study also indicates that G. fossarum might be capable of dispersing overland in headwater regions and of crossing small in‐stream barriers. 4. We speculate that differences in dispersal capacity but also habitat specialisation and potentially the extent of local adaptation could be responsible for the differences in genetic differentiation found between the two species. These results highlight the importance of taking into account dispersal‐related traits when planning management and conservation strategies.     

Staphylorchis cymatodes (Gorgoderidae: Anaporrhutinae) from carcharhiniform, orectolobiform and myliobatiform elasmobranchs of Australasia: Low host specificity, wide distribution and morphological plasticity

Anaporrhutine gorgoderids (Digenea: Gorgoderidae: Anaporrhutinae) found in the body cavity of six species of elasmobranchs from the orders Carcharhiniformes, Myliobatiformes and Orectolobiformes from Australian waters were found to belong to the genus Staphylorchis. Although these specimens were morphologically variable, sequences of ITS2 and 28S ribosomal DNA from specimens from three host families and two host orders were identical. Based on morphological and molecular data these specimens were identified as the type-species of the genus, Staphylorchis cymatodes. New measurements are provided for S. cymatodes, and for the first time genetic data are presented for this species. In addition to providing new morphological and molecular data for S. cymatodes, the previously described species S. gigas, S. parisi and S. scoliodonii, are here synonymised with S. cymatodes. This implies that S. cymatodes, as conceived here, has remarkably low host-specificity, being recorded from eight elasmobranch species from four families and three orders, has a wide geographical distribution in the Indo-west Pacific from off India, in the Bay of Bengal, to Moreton Bay in the Coral Sea, and is morphologically plastic, with body size, size of specific organs and body shape differing dramatically between specimens from different host species. The genus Staphylorchis now contains only two valid species, S. cymatodes and S. pacifica.     

Integrated Taxonomy Reveals Hidden Diversity in Northern Australian Fishes: A New Species of Seamoth (Genus Pegasus)

Fishes are one of the most intensively studied marine taxonomic groups yet cryptic species are still being discovered. An integrated taxonomic approach is used herein to delineate and describe a new cryptic seamoth (genus Pegasus) from what was previously a wide-ranging species. Preliminary mitochondrial DNA barcoding indicated possible speciation in Pegasus volitans specimens collected in surveys of the Torres Strait and Great Barrier Reef off Queensland in Australia. Morphological and meristic investigations found key differences in a number of characters between P. volitans and the new species, P. tetrabelos. Further mt DNA barcoding of both the COI and the slower mutating 16S genes of additional specimens provided strong support for two separate species. Pegasus tetrabelos and P. volitans are sympatric in northern Australia and were frequently caught together in trawls at the same depths.     

Stem CO2 efflux of ten species in temperate forests in Northeastern China

Stem CO₂ efflux (E _S) is an important component of forest ecosystem carbon budgets and net ecosystem CO₂ exchange, but little is known about E _S in temperate forests in Northeastern China, an area with a large extent of forest. We measured E _S along with stem temperature at 1?cm depth (Ts) over a 9?month period in 2007 on ten dominant tree species of secondary forests of the region. Other measurements included the autotrophic component of soil CO₂ efflux (E _A) and stem diameter at breast height (DBH). Our objectives were to (1) examine the seasonal patterns and species differences in E _S, and (2) determine the correlations between E _S and Ts, DBH and E _A. Mean E _S for the measurement period ranged from 1.09 to 1.74?μmol?CO₂?m^?2?s^?1 among the ten species. The sensitivity of E _S to Ts (Q ₁₀ ) ranged from 1.87 to 2.61. Across the ten species 57–89% of variation in E _S was explained by T _S and DBH. There was also a linear relationship between mean E _S and E _A. E _S was better predicted by Ts in the dormant season than the growing season, indicating that additional factors such as growth respiration and internal transport of CO₂ in the xylem became more important contributors to E _S during the growing season. Stem CO₂ efflux increased, and Q ₁₀ decreased, with increasing DBH in all species. Although temperature exerts strong control on the rate of cellular respiration, we conclude that in tree stems in situ, T _S, DBH and many other factors affect the relationship between CO₂ evolution by respiring cells and the diffusion of CO₂ to the stem surface.     

Fungi are involved in the effects of litter mixtures on consumption by shredders

1. Decomposition of litter mixtures in both terrestrial and aquatic ecosystems often shows non‐additive diversity effects on decomposition rate, generally interpreted in streams as a result of the feeding activity of macroinvertebrates. The extent to which fungal assemblages on mixed litter may influence consumption by macroinvertebrates remains unknown. 2. We assessed the effect of litter mixing on all possible three‐species combinations drawn from four tree species (Alnus glutinosa, Betula pendula, Juglans regia and Quercus robur) on both fungal assemblages and the rate of litter consumption by a common shredder, Gammarus fossarum. After a 9‐week inoculation in a stream, batches of leaf discs were taken from all leaf species within litter mixture combinations. Ergosterol, an indicator of fungal biomass, and the composition of fungal assemblages, assessed from the conidia released, were determined, and incubated litter offered to G. fossarum in a laboratory‐feeding experiment. 3. Mixing leaf litter species enhanced both the Simpson’s index of the fungal assemblage and the consumption of litter by G. fossarum, but had no clear effect on mycelial biomass. Specifically, consumption rates of J. regia were consistently higher for mixed‐species litter packs than for single‐species litter. In contrast, the consumption rates of B. pendula were not affected by litter mixing, because of the occurrence of both positive and negative litter‐mixing effects in different litter species combinations that counteracted each other. 4. In some litter combinations, the greater development of some fungal species (e.g. Clavariopsis aquatica) as shown by higher sporulation rates coincided with increased leaf consumption, which may have resulted from feeding preferences by G. fossarum for these fungi. 5. Where litter mixture effects on decomposition rate are mediated via shredder feeding, this could be due to indirect effects of the fungal assemblage.     

Life history and reproductive capacity of Gammarus fossarum and G. roeseli (Crustacea: Amphipoda) under naturally fluctuating water temperatures: a simulation study

SUMMARY 1. Mathematical functions developed in long‐term laboratory experiments at different constant temperatures were combined with daily water temperatures for 1991–93 in eight Austrian streams and rivers to simulate the complex life histories and reproductive capacities of two freshwater amphipods: Gammarus fossarum and G. roeseli. The functions describe brood development times, hatching success, times taken to reach sexual maturity, growth, and fecundity. The sex ratio was assumed to be 0.5 and an autumn–winter reproductive resting period was based on observations of six river populations. Simulations included summer‐cold mountain streams, summer‐warm lowland rivers, watercourses fed by groundwater or influenced by heated effluents, and varying amplitudes of change within each year. 2. A fortran 77 computer program calculated growth from birth to sexual maturity of first‐generation females born on the first day of each calendar month in 1991, and the numbers of offspring successfully released from the maternal broodpouch in successive broods. At the 1991–93 regimes of temperature, individual G. fossarum released 127–208 offspring and G. roeseli released 120–169 in seven or eight successive broods during life spans of less than 2 years in six rivers. Life spans extended into a third year in the relatively cool River Salzach (mean temperature 7.5 °C). They were not completed in the very cold River Steyr (mean 5.6, range 2.5–7.9 °C), where G. fossarum produced five broods (totalling 120 offspring) and G. roeseli only two broods (totalling 28 offspring) in the 3‐year period. Except in the Steyr, some offspring grew rapidly to maturity and produced several second‐generation broods during the simulation period; in the warmest rivers some third‐generation broods were also produced. Birth dates, early or late in the year, influenced the subsequent production of broods and young, depending on temperature regimes in particular rivers. Total numbers of offspring produced by the second and third generations represent the theoretical reproductive capacities of G. fossarum and G. roeseli. Minimum and maximum estimates mostly ranged from 100 to 17 300, were larger for G. fossarum except in the warmest river (March), where temperatures rose above 20 °C for 56–78 days in summer, and largest (maximum 37 600) in the River Voeckla heated by discharge from a power‐station (mean 11.5 °C). Results from the simulations agree with preliminary assessments of relative abundances for G. fossarum and G. roeseli in several of the study rivers, but in some one or both species appear to be absent. On a wider scale, the present study confirms that G. fossarum is potentially more successful than G. roeseli in cool rivers but indicates that neither species is likely to maintain viable populations in cold rivers strongly influenced by snow and ice‐melt. 3. The potential impacts of future river warming by increases of 1, 2 and 3 °C, due to climate change, vary according to river site, date of fertilisation, the extent of temperature increase, and the species of Gammarus. For Austrian rivers with mean temperatures in the range c. 7–10 °C, future warming would result in modest changes in the life histories and reproductive capacities of both G. fossarum and G. roeseli; the former would find improved temperature conditions in watercourses that are currently very cold throughout the year, and both would find warm rivers less tolerable. 4. The high potential reproductive capacity of gammarids, with rapid production of numerous successive broods when sexual maturity is finally achieved, indicates adaptation to high mortality during the relatively long period of growth to sexual maturity, and provides scope for an opportunistic strategy of emigration from centres of population abundance to colonise new territory when conditions are favourable. Rapid expansion of populations is desirable to combat the effects of environmental catastrophes, both frequent and short‐term floods and droughts, and more long‐term climatic changes that have occurred several times in glacial–interglacial periods during the current Ice Age.     

Population structure and cryptic genetic variation in the mango fruit fly,Ceratitis cosyra (Diptera,Tephritidae)

The fruit fly Ceratitis cosyra is an important agricultural pest negatively affecting the mango crop production throughout Africa and also feeding on a variety of other wild and cultivated hosts. The occurrence of deeply divergent haplotypes, as well as extensive morphological variability, previously suggested possible cryptic speciation within Ceratitis cosyra. Here we provide the first large-scale characterisation of the population structure of Ceratitis cosyra with the main objective of verifying cryptic genetic variation. A total of 348 specimens from 13 populations were genotyped at 16 polymorphic microsatellite loci. Hardy-Weinberg equilibrium (HWE) deviations were observed in 40.4% of locus-population combinations and suggested the occurrence of genetic substructuring within populations. Discriminant Analysis of Principal Components (DAPC) showed genetic divergence between the vast majority of vouchers from Burundi and Tanzania (plus a few outliers from other African countries) and all other specimens sampled. Individual Bayesian assignments confirmed the existence of two main genotypic groups also occurring in sympatry. These data provided further support to the hypothesis that Ceratitis cosyra might include cryptic species. However, additional integrative taxonomy, possibly combining morphological, ecological and physiological approaches, is required to provide the necessary experimental support to this model.     

A Microbial Link between Elevated CO2 and Methane Emissions that is Plant Species-Specific

Rising atmospheric CO₂ levels alter the physiology of many plant species, but little is known of changes to root dynamics that may impact soil microbial mediation of greenhouse gas emissions from wetlands. We grew co-occurring wetland plant species that included an invasive reed canary grass (Phalaris arundinacea L.) and a native woolgrass (Scirpus cyperinus L.) in a controlled greenhouse facility under ambient (380 ppm) and elevated atmospheric CO₂ (700 ppm). We hypothesized that elevated atmospheric CO₂ would increase the abundance of both archaeal methanogen and bacterial methanotroph populations through stimulation of plant root and shoot biomass. We found that methane levels emitted from S. cyperinus shoots increased 1.5-fold under elevated CO₂, while no changes in methane levels were detected from P. arundincea. The increase in methane emissions was not explained by enhanced root or shoot growth of S. cyperinus. Principal components analysis of the total phospholipid fatty acid (PLFA) recovered from microbial cell membranes revealed that elevated CO₂ levels shifted the composition of the microbial community under S. cyperinus, while no changes were detected under P. arundinacea. More detailed analysis of microbial abundance showed no impact of elevated CO₂ on a fatty acid indicative of methanotrophic bacteria (18:2ω6c), and no changes were detected in the terminal restriction fragment length polymorphism (T-RFLP) relative abundance profiles of acetate-utilizing archaeal methanogens. Plant carbon depleted in ¹³C was traced into the PLFAs of soil microorganisms as a measure of the plant contribution to microbial PLFA. The relative contribution of plant-derived carbon to PLFA carbon was larger in S. cyperinus compared with P. arundinacea in four PLFAs (i14:0, i15:0, a15:0, and 18:1ω9t). The δ¹³C isotopic values indicate that the contribution of plant-derived carbon to microbial lipids could differ in rhizospheres of CO₂-responsive plant species, such as S. cyperinus in this study. The results from this study show that the CO₂–methane link found in S. cyperinus can occur without a corresponding change in methanogen and methanotroph relative abundances, but PLFA analysis indicated shifts in the community profile of bacteria and fungi that were unique to rhizospheres under elevated CO₂.     

Sibling species of fragrant orchids (Gymnadenia: Orchidaceae, Magnoliophyta) in Russia

Recent molecular phylogenetic studies of the genus Gymnadenia have demonstrated that it contains sibling taxa, i.e., species that are hardly distinguishable according to morphological traits, yet are phylogenetically rather distant and distinctly distinguishable by molecular methods, which is a rare phenomenon for angiosperms. By sequencing the ITS1-5.8S rDNA-ITS2 fragment the presence of G. densiflora was confirmed for Russia. Our data supported a high degree of genetic differentiation between G. conopsea s. str. and G. densiflora, which confirms their taxonomic rank of species. Morphological analysis has shown that the features that are best for the discrimination between these two species in Northwestern Russia are the length of the lower bract, length of the mid-lobe of the lip, and width of leaves. The ecological and phenological differentiation between G. conopsea s. str. and G. densiflora is briefly reviewed. The ITS sequence variation in these species has been analyzed; the molecular genetic differences of the G. conopsea individuals from the eastern part of the distribution area have been discovered for the first time. Different taxonomic interpretations of Gymnadenia phylogenetic tree topology taking into account the presence of sibling species are discussed in general.     

A biophysical basis for the emergence of the genetic code in protocells

The origin of the genetic code is an abiding mystery in biology. Hints of a ‘code within the codons’ suggest biophysical interactions, but these patterns have resisted interpretation. Here, we present a new framework, grounded in the autotrophic growth of protocells from CO₂ and H₂. Recent work suggests that the universal core of metabolism recapitulates a thermodynamically favoured protometabolism right up to nucleotide synthesis. Considering the genetic code in relation to an extended protometabolism allows us to predict most codon assignments. We show that the first letter of the codon corresponds to the distance from CO₂ fixation, with amino acids encoded by the purines (G followed by A) being closest to CO₂ fixation. These associations suggest a purine-rich early metabolism with a restricted pool of amino acids. The second position of the anticodon corresponds to the hydrophobicity of the amino acid encoded. We combine multiple measures of hydrophobicity to show that this correlation holds strongly for early amino acids but is weaker for later species. Finally, we demonstrate that redundancy at the third position is not randomly distributed around the code: non-redundant amino acids can be assigned based on size, specifically length. We attribute this to additional stereochemical interactions at the anticodon. These rules imply an iterative expansion of the genetic code over time with codon assignments depending on both distance from CO₂ and biophysical interactions between nucleotide sequences and amino acids. In this way the earliest RNA polymers could produce non-random peptide sequences with selectable functions in autotrophic protocells.     

Reproductive potential and lifetime potential fecundity of the freshwater amphipods Gammarus fossarunt and G. roeseli in Austrian streams and rivers

• 1 Fecundity of Gammarus fossarum populations at six stream sites (S1-S6) and G. roeseli populations at two sites (S5 and S6) was studied monthly during 1985–1988, distinguishing seven morphological stages of embryonic development. Mean survival of G. fossarum eggs was 65% (60–70%) in nature and 60% (57–63%) in the laboratory; egg survival of G. roeseli was 52% (46–58%) in nature and 41% (35–47%) in the laboratory. Regressions of egg numbers per clutch against body wet weight (WWT) decreased significantly for egg developmental stages 2 to 7 (hatching).
• 2 In stream populations, ovigerous females of G. fossarum were absent in October (six sites) and November (five sites); ovigerous G. roeseli were usually absent from October until March.
• 3 Overall mean egg volume (EV) increased significantly from 0,08 ± 0.001 mm³ and 0.075 ± 0.001 mm³ for stage 2 eggs, to 0.174 ± 0.012mm³ and 0.160 ± 0.013 mm³ for stage 6 eggs of G. fossarum and G. roeseli, respectively. The dry weights of stage 2 eggs, stage 6 eggs and neonates (stage 7) were not significantly different, within and between species; mean dry weight was 36 ± 3 μg.
• 4 Egg volumes and fecundity indices (FI, weight-specific number of eggs per clutch) were not significantly different between the same months of four successive years, so the data were pooled to give a single representative ‘year’. At most sites there were marked seasonal fluctuations in these variables. In both species, mean EV was largest for ‘winter’ eggs in December/January and smallest for ‘summer’ eggs in May, EV decreased during some months of the year when FI increased, and vice versa. Mean reproductive effort (RE = EV × FI) declined from high ‘winter’ values to low ‘summer’ values, and this difference was most marked at sites where the major foodstuff available was seasonally limited (autumn-shed tree leaves). Overall (‘annual’) mean RE was also lowest at these sites (S3 and S4) but was some 13% greater at sites where nutrients and food were present in the greatest quantifies (S5 and S6). Specific maximum potential body sizes and hence maximum potential fecundities (fifty-nine eggs for G. fossarum and eighty-seven eggs for G. roeseli) were attained only at S5 and 56. Mean RE was nearly identical for G. fossarum and G. roeseli from the same sites.
• 5 At experimental temperatures (T) in the range 3.8–20.2°C, females of the same body weight carried significantly different egg numbers per clutch. The relationship between FI and T was described by a second-order polynomial. The calculated optimum T was 12.1°C for G. fossarum and 16.3°C for G. roeseli. Interspecific differences were highly significant.
• 6 Theoretical lifetime potential fecundity was approximately 194 eggs for G. fossarum

     

Performance of distance-based DNA barcoding in the molecular identification of Primates

For comparative primatology proper recognition of basal taxa (i.e. species) is indispensable, and in this the choice of a suitable gene with high phylogenetic resolution is crucial. For the goals of species identification in animals, the cytochrome c oxidase subunit 1 (cox1) has been introduced as standard marker. Making use of the difference in intra- and interspecific genetic variation – the DNA barcoding gap – cox1 can be used as a fast and accurate marker for the identification of animal species. For the Order Primates we compare the performance of cox1 (166 sequences; 50 nominal species) in species-identification with that of two other mitochondrial markers, 16S ribosomal RNA (412 sequences, 92 species) and cytochrome b (cob: 547 sequences, 72 species). A wide gap exist between intra- and interspecific divergences for both cox1 and cob genes whereas this gap is less apparent for 16S, indicating that rRNA genes are less suitable for species delimitation in DNA barcoding. For those species where multiple sequences are available there are significant differences in the intraspecific genetic distances between different mitochondrial markers, without, however, showing a consistent pattern. We conclude that cox1 allows accurate differentiation of species and as such DNA barcoding may have an important role to play in comparative primatology.     

Genotypes and subtypes of Cryptosporidium spp. in diarrheic lambs and goat kids in northern Greece

Inconsistent data exist on the distribution of zoonotic Cryptosporidium species and subtypes in sheep and goats in European countries, and few such data are available from Greece. In this study, 280 fecal specimens were collected from 132 diarrheic lambs and 148 diarrheic goat kids aged 4 to 15?days on 15 farms in northern Greece, and examined for Cryptosporidium spp. using microscopy of Ziehl-Neelsen-stained fecal smears. Cryptosporidium spp. in 80 microscopy-positive fecal specimens (39 from lambs and 41 from goat kids) were genotyped by PCR-RFLP analysis of the small subunit rRNA gene and subtyped by sequence analysis the 60?kDa glycoprotein gene. Among the 33 specimens successfully genotyped, C. parvum was found in 32 and C. xiaoi in one. Seven subtypes belonging to two subtype families (IIa and IId) were identified among the 29 C. parvum specimens successfully subtyped, including IIaA14G2R1 (1/29), IIaA15G2R1 (6/29), IIaA20G1R1 (7/29), IIdA14G2 (1/29), IIdA15G1 (9/29), IIdA16G1 (3/29), and IIdA23G1 (2/29). Lambs were more commonly infected with C. parvum IIa subtypes, whereas goat kids were more with IId subtypes. The results illustrate that C. parvum is prevalent in diarrheic lambs and goat kids in northern Greece and these animals could potentially play a role in epidemiology of human cryptosporidiosis.     

Variation in fish community structure, richness, and diversity in 56 Danish lakes with contrasting depth, size, and trophic state: does the method matter?

Organisms inhabiting shallow near-shore waters are at risk of desiccation during water level fluctuations. Using laboratory experiments, we investigated the survival and behavioural defences of four freshwater amphipod species during substratum drying: three Ponto-Caspian invaders (Pontogammarus robustoides, Dikerogammarus haemobaphes and Dikerogammarus villosus) and the native Gammarus fossarum. We hypothesized that they would be able to survive air exposure events as well as to adjust their behaviour by following the decreasing water level and/or burying in the sediments. To test these hypotheses, we examined survival of each species on gradually drying sandy substratum as well as their horizontal and vertical migration behaviours. P. robustoides was most resistant to substratum drying and was the only species burying into the substratum. On the other hand, G. fossarum exhibited distinct horizontal migrations following the retreating waterline. These two species seem to be particularly well adapted to the drying environment. Defence mechanisms of D. haemobaphes and D. villosus were less efficient, though the former species also followed the retreating waterline to some extent. Our study demonstrates that exotic and native gammarids have several adaptations that enable them to invade and persist in habitats experiencing common water level fluctuations.