Stonefly Species Research Articles

<strong>A study of stonefly taxa (Insecta: Plecoptera) inhabiting a saline stream (Arroyo Salado) in southern Spain</strong>

Few studies of the ecology of Mediterranean saline streams exist. The strong selective pressure exerted by salinity greatly limits the presence of many taxa, Plecoptera (stoneflies) among them. However, a few species of stoneflies have been reported inhabiting saline streams, particularly components of the thermophilous biocoenosis seemed to have adapted to salinity pulses caused by fluctuations in annual rainfall. For two years, three sites within a saline stream, Arroyo Salado in southern Iberian Peninsula, were sampled to determine the stonefly species composition. Conductivity at the three sites ranged from 2290 to 87792, 2550 to 90824, and 206 to 2818 μS/cm, respectively. A total of seven different stonefly taxa were identified at the three sites: Brachyptera auberti Consiglio, Nemoura lacustris Pictet, Capnioneura libera (Navás), Tyrrhenoleuctra sp., Hemimelaena flaviventris (Pictet), Protonemura alcazaba Aubert, and Perla marginata (Panzer), the latter two only at the site where the stream flowed continuously and conductivity was lower. The possible strategies enabling these stonefly species to surviving in Arroyo Salado are discussed.

<strong>Toward a comprehensive COI DNA barcode library for Swiss Stoneflies (Insecta: Plecoptera) with special emphasis on the genus <em>Leuctra</em></strong>

The Swiss Barcode of Life initiative (SwissBOL) aims to inventory the genetic biodiversity in Switzerland using a short DNA sequence. DNA barcoding provides an additional tool for species identification that complements traditional morphological approaches. We report on the establishment of a DNA barcode library for Plecoptera, taxa that are of great importance as bioindicators of water quality and that often present difficulties in species-level identification for larvae and female specimens. Non-destructive DNA extraction, PCR amplification and sequencing of part of the mitochondrial gene Cytochrome Oxidase I (COI) was conducted for 440 individuals (one to eight per species) belonging to 90 species (of the 112 reported from Switzerland). Intra and interspecific distances were calculated and gene trees reconstructed. In most cases, COI was efficient in delimiting stonefly species. Some doubtful specimens were subsequently re-examined and a few misidentifications were found, especially in some problematic groups in the genus Leuctra Stephens, 1836. Larger genetic distances in some species (e.g. Leuctra nigra (Olivier 1811)) indicate the possible presence of sibling species, while in a few cases closely related species are genetically difficult to separate (within the Leuctra fusca species group).

<strong>Contribution to the taxonomy and biology of two Balkan endemic <em>Isoperla</em> Banks, 1906 (Plecoptera: Perlodidae) species</strong>

We contribute new information on the taxonomy and biology of two recently described Balkan endemic stonefly species of the genus Isoperla Banks, 1906 (Plecoptera: Perlodidae). The hitherto unknown larva of Isoperla pesici Murányi, 2011 can be distinguished from related species on the basis of more laterally positioned abdominal stripes, and the long hairs on the apical half of the cerci. This species has a wide distribution in the Central Balkans, and we add six new localities, including the first record from Albania. It emerges from late spring to mid-summer, and occurs at low to moderately high altitudes. Isoperla pesici seems to be associated with springs, especially karst spring outlets, although it occurs in both small and large springs. The hitherto unknown larva of Isoperla citrina Murányi, 2011 is distinguished by its brown coloration with an indistinct pattern, quadrangular lacinia and lack of long hairs on the apical half of the cerci. The female lacks distinctive terminalia or egg characters and its habitus is similar to the male and sufficient to distinguish it from other known Balkan taxa. The male mating call is a sequence of beat-groups with a total duration of 3–6 s (at 22 °C); each group generally containing 3 beats, while the beat repetition pattern within beat groups is rather simple. This species is restricted to the high mountains of the western-Central Balkans, where we record four new localities, including the first record from Macedonia. It emerges from June to October, and is associated with very small creeks, lined with bushes at high elevations. Amongst the accompanying species, eleven are reported for the first time from Albania: (Brachyptera helenica Aubert, 1956, B. graeca Berthélemy, 1971, Leuctra bronislawi Sowa, 1970a, L. hirsuta Bogoecu & Tabacaru, 1960, L. hippopoides Kaçanski & Zwick, 1970, L. pseudosignifera Aubert, 1954, Protonemura hrabei Raušer, 1956, P. nitida (Pictet, 1836), Nemurella pictetii (Klapálek, 1900), Chloroperla russevi Braasch, 1969 and Siphonoperla neglecta (Rostock, 1881)), while one from Macedonia (Nemoura caligula Zwick, 1978) and Leuctra pseudohippopus Raušer, 1965 is reported from outside the Moesian ranges for the first time.

The first fossil salmonfly (Insecta: Plecoptera: Pteronarcyidae), back to the Middle Jurassic.

BackgroundThe fossil record of Plecoptera (stoneflies) is considered relatively complete, with stem-groups of each of the three major lineages, viz. Antarctoperlaria, Euholognatha and Systellognatha (and some of their families) represented in the Mesozoic. However, the family Pteronarcyidae (the salmonflies; including two genera, Pteronarcys and Pteronarcella) has no fossil record to date, and the family has been suggested to have diverged recently.ResultsIn this paper, we report on a set of specimens belonging to a new fossil species of stonefly, discovered from the Middle Jurassic Daohugou locality (China). Our comparative analysis of wing venation and body characters demonstrates that the new species belongs to the Pteronarcyidae, and is more closely related to Pteronarcys than to Pteronarcella. However, it differs from all known species of the former genus. It is therefore assigned to a new genus and named Pteroliriope sinitshenkovae gen. et sp. nov. under the traditional nomenclatural procedure. The cladotypic nomenclatural procedure is also employed, with the resulting combination Pteroliriope nec Pteronarcys sinitshenkovae sp. nov. ConclusionsThe first discovery of a fossil member of the Pteronarcyidae demonstrates that the corresponding lineage is not a very recent offshoot but was already present ca. 165 million years ago. This discovery concurs with the view that divergence of most stonefly families took place very early, probably in the Triassic, or even in the Permian. This contribution demonstrates the need for (re-)investigations of the systematics of fossil stoneflies to refine divergence date estimates for Plecoptera lineages.

A remarkable new species of winter stonefly (Plecoptera: Capniidae) from Southeastern China.

A new species of the genus Capnia s.l., C. zijinshana Du & Chen, sp. nov. is described and illustrated from Jiangsu Province, of southeastern China. The new species is distinguished from other known Chinese capniid species by the unique thoracic sclerites and genitalic characters.

Feeding and parasitism of adultStenoperlaspp. (Plecoptera: Eustheniidae) in New Zealand

AbstractThe adult diets of two species of stonefly,Stenoperla prasina(Newman, 1845) andStenoperla maclellaniZwick, 1979 were investigated by gut content analysis. Adults were collected throughout New Zealand by light‐trapping at 77 streamside sites most of which were located in native forest.S. prasinawas widely distributed and collected at 43 sites, whereasS. maclellaniwas sampled at 10 sites in the South Island only. Of 200 S. prasinaand 42 S. maclellanidissected, 36% and 69% respectively, contained food. Microscopic examination of a subset of individuals indicated that the gut contents of both species were dominated by sooty mould fungi with lesser contributions made by plant tissue, pollen grains and the spores of tree ferns. These findings suggest that stoneflies had been feeding on the trunks, branches and foliage of trees and perhaps on fronds of tree ferns. As the abdomens of most females were packed with eggs, which mature in late‐instar nymphs it is likely that feeding provides adults with a source of immediately available energy for mating and flight. Encysted larval stages of hairworms (Nematomorpha) and a trematode were also found in both stonefly species. Metacercarial cysts of the trematode are the first records of larval flukes parasitising New Zealand stoneflies. They were found in about 43% of dissected adults of each species with up to 40 cysts present in some individuals. It is possible that the parasite's definitive host is one of New Zealand's native bats (Chiroptera), which are known to eatStenoperlaand inhabit streamside forests at some of the localities where parasitised stoneflies were taken. Overall, our results provide further evidence for the importance of sooty mould fungi as a food source for New Zealand insects and highlight the role ofStenoperlaas a vector of larval parasites from aquatic to terrestrial hosts.

Loss of Genetic Diversity and Increased Subdivision in an Endemic Alpine Stonefly Threatened by Climate Change.

Much remains unknown about the genetic status and population connectivity of high-elevation and high-latitude freshwater invertebrates, which often persist near snow and ice masses that are disappearing due to climate change. Here we report on the conservation genetics of the meltwater stonefly Lednia tumana (Ricker) of Montana, USA, a cold-water obligate species. We sequenced 1530 bp of mtDNA from 116 L. tumana individuals representing “historic” (>10 yr old) and 2010 populations. The dominant haplotype was common in both time periods, while the second-most-common haplotype was found only in historic samples, having been lost in the interim. The 2010 populations also showed reduced gene and nucleotide diversity and increased genetic isolation. We found lower genetic diversity in L. tumana compared to two other North American stonefly species, Amphinemura linda (Ricker) and Pteronarcys californica Newport. Our results imply small effective sizes, increased fragmentation, limited gene flow, and loss of genetic variation among contemporary L. tumana populations, which can lead to reduced adaptive capacity and increased extinction risk. This study reinforces concerns that ongoing glacier loss threatens the persistence of L. tumana, and provides baseline data and analysis of how future environmental change could impact populations of similar organisms.

Introgression and species demarcation in western European Leuctra fusca (Linnaeus, 1758) and L. digitata Kempny, 1899 (Plecoptera: Leuctridae)

ABSTRACTThe compilation of a DNA barcoding library of Norwegian stonefly (Plecoptera) species revealed that Leuctra fusca (Linnaeus, 1758) and Leuctra digitata Kempny, 1899 (Leuctridae) share haplotypes in northernmost Scandinavia. Phylogenetic analyses of the mitochondrial (mt) DNA barcode marker COI and the nuclear marker 28S show that the shared haplotypes must result from the introgression of a L. fusca mitochondrion into a L. digitata population on at least two occasions. Although mt introgression is widespread in animals, this represents the first documented case in Plecoptera. This study also included specimens of L. cf. fusca from the Sierra Nevada massif in Spain, a population previously known as L. carpentieri Despax, 1945. Their mt haplotypes are ca. 13% different from other European L. fusca. However, their 28S alleles are compatible with their morphological identification as L. fusca. In view of the possibility of mt introgression, the taxonomic status of this population remains undecided.

The stoneflies (Plecoptera) of Belarus: Fauna and biotopic distribution

A total of 29 species of stoneflies (Plecoptera) from 7 families were found in Belarus: Perlodidae—6, Perlidae—1, Chloroperlidae—3, Taeniopterygidae—2, Nemouridae—11, Capniidae—2, and Leuctridae—4 species. The fauna is fairly diverse and includes a number of rare species in Belarus and other European countries: Diura bicaudata, Marthamea vitripennis, Isoptena serricornis, Xanthoperla apicalis, Brachyptera risi, Protonemura intricata, Nemoura cambrica, Nemoura marginata, and Capnia bifrons.

New species records of systellognathan stoneflies (Plecoptera: Systellognatha) for the fauna of the Korean Peninsula

Three species of stoneflies are reported for the first time from the Korean Peninsula: Pictetiella asiatica Zwick & Levanidova, 1971, Xanthoneuria unimaculata (Zhiltzova, 1979) and a new species of the genus Sweltsa Ricker, 1943. The widespread temperate Asian species, P. asiatica, represents the first record of the Transpacific genus Pictetiella Illies, 1966 from Korea. Xanthoneuria unimaculata confirms the presence of Xanthoneuria Uchida, 2011 in Korea. This genus was hitherto known only from Japan and the Russian Far East with a dubious record from Korea. The female of Sweltsa sp. is formally described but not named becaus e male remained unknown. All the specimen s were collected in North Korea by colleagues from the Hungarian Natural History Museum duri ng the 1970’s and 1980’s. New Korean records of further ten species are also enumerated, with notes about some doubtful records. Distribution area of the genera Pictetiella and Xanthoneuria are detailed and depicted on maps.

A new genus and species of winter stoneflies (Plecoptera: Capniidae) from Southwest China, with a commented checklist of the family in the Oriental Realm

A new genus and species of the family Capniidae (Plecoptera), Sinocapnia kuankuoshui gen. n., sp. n. is described from the adult male and a female collected in Guizhou Province of southwest China. The new taxon is distinguished from all extant Capniidae genera and the assemblage of species currently included in Capnia sensu lato by combination of unique genitalic, wing, and thoracic sclerites characters. No closely related taxon is indicated on the basis of morphological characters. An annotated checklist of the family Capniidae from the Oriental Realm is given.

Genome-wide SNPs reveal fine-scale differentiation among wingless alpine stonefly populations and introgression between winged and wingless forms

Insect flight loss is a repeated phenomenon in alpine habitats, where wing reduction is thought to enhance local recruitment and increase fecundity. One predicted consequence of flight loss is reduced dispersal ability, which should lead to population genetic differentiation and perhaps ultimately to speciation. Using a dataset of 15,123 SNP loci, we present comparative analyses of fine-scale population structure in codistributed Zelandoperla stonefly species, across three parallel altitudinal transects in New Zealand's Rock and Pillar mountain range. We find that winged populations (altitude 200-500 m; Zelandoperla decorata) show no genetic structuring within or among streams, suggesting substantial dispersal mediated by flight. By contrast, wingless populations (Zelandoperla fenestrata; altitude 200-1100 m) exhibit distinct genetic clusters associated with each stream, and additional evidence of isolation by distance within streams. Our data support the hypothesis that wing-loss can initiate diversification in alpine insect populations over small spatial scales. The often deep phylogenetic placement of lowland Z. fenestrata within their stream-specific clades suggests the possibility of independent alpine colonization events for each stream. Additionally, the detection of winged, interspecific hybrid individuals raises the intriguing possibility that a previously flightless lineage could reacquire flight via introgression.

Ecological Effects of Biochar on the Structure and Function of Stream Benthic Communities

The introduction of biochar, activated carbon, and other carbonaceous materials to aquatic ecosystems significantly reduces the toxicity and bioavailability of contaminants. However, previous studies have shown that these materials can have negative effects on aquatic organisms. We conducted field and mesocosm experiments to test the hypothesis that biochar altered the structure and function of stream benthic communities. After 30 d in the field, colonization by stoneflies (Plecoptera) was significantly lower in trays containing biochar compared to the results from the controls. In stream mesocosms, biochar increased macroinvertebrate drift and significantly reduced community metabolism. However, most measures of community composition showed little variation among biochar treatments, and significant responses were limited to a single stonefly species (Capnia confusa). When benthic communities were simultaneously exposed to biochar and Cu, effects were primarily associated with metal exposure. Because it is unlikely that biochar treatments would be employed in uncontaminated areas, these moderately negative effects should be considered within the context of the positive benefits associated with reduced contaminant bioavailability and toxicity. Additional research is necessary to improve our understanding of the mechanisms responsible for biochar effects on benthic communities and to identify the optimal application rates and size fractions that will maximize contaminant sorption but minimize potential negative effects.

Can DNA-Based Ecosystem Assessments Quantify Species Abundance? Testing Primer Bias and Biomass--Sequence Relationships with an Innovative Metabarcoding Protocol.

Metabarcoding is an emerging genetic tool to rapidly assess biodiversity in ecosystems. It involves high-throughput sequencing of a standard gene from an environmental sample and comparison to a reference database. However, no consensus has emerged regarding laboratory pipelines to screen species diversity and infer species abundances from environmental samples. In particular, the effect of primer bias and the detection limit for specimens with a low biomass has not been systematically examined, when processing samples in bulk. We developed and tested a DNA metabarcoding protocol that utilises the standard cytochrome c oxidase subunit I (COI) barcoding fragment to detect freshwater macroinvertebrate taxa. DNA was extracted in bulk, amplified in a single PCR step, and purified, and the libraries were directly sequenced in two independent MiSeq runs (300-bp paired-end reads). Specifically, we assessed the influence of specimen biomass on sequence read abundance by sequencing 31 specimens of a stonefly species with known haplotypes spanning three orders of magnitude in biomass (experiment I). Then, we tested the recovery of 52 different freshwater invertebrate taxa of similar biomass using the same standard barcoding primers (experiment II). Each experiment was replicated ten times to maximise statistical power. The results of both experiments were consistent across replicates. We found a distinct positive correlation between species biomass and resulting numbers of MiSeq reads. Furthermore, we reliably recovered 83% of the 52 taxa used to test primer bias. However, sequence abundance varied by four orders of magnitudes between taxa despite the use of similar amounts of biomass. Our metabarcoding approach yielded reliable results for high-throughput assessments. However, the results indicated that primer efficiency is highly species-specific, which would prevent straightforward assessments of species abundance and biomass in a sample. Thus, PCR-based metabarcoding assessments of biodiversity should rely on presence-absence metrics.

The nearly complete mitochondrial genome of a stonefly species, Styloperla sp. (Plecoptera: Styloperlidae)

We report the nearly complete mitochondrial genome of a stonefly species, Styloperla sp. (Plecoptera: Styloperlidae), which is a circular molecule of 15,416 bp in length and consists of 13 protein-coding genes, 2 ribosomal RNAs, 20 transfer RNAs and a partial control region (645 bp). Using the 13 protein-coding genes of 8 stoneflies and 3 other related species, we constructed a phylogenetic tree to verify the accuracy of the new determined mitogenome sequences. Our results provide basic data for further study of phylogeny in Plecoptera.

Life cycles and production of three predatory stonefly species (Plecoptera, Insecta) in the Kedrovaya River (Southern Primorskii Territory)

The life cycles and production of populations of the predatory stoneflies Skwala compacta, Stavsolus manchuricus, and Kamimuria exilis were studied in the Kedrovaya salmon stream (the Kedrovaya Pad Reserve, the south of Primorskii Territory). Skwala compacta and S. manchuricus are univoltine with direct embryonic development and slow nymphal growth during 311 and 294 days, respectively, including winter growth at low water temperatures. The life history of S. compacta was characterized by synchronous emergence and population recruitment, an uninterrupted increase in the nymphal body weight from May to September, rapid growth in October–November, and a sustained weight gain during the winter months until emergence in April–May. The life history of S. manchuricus was characterized by an extended period of emergence in May–July and recruitment from July to September, as well as by a wide range of size-age parameters, presumably related to the specificity of the reproductive system in adults and an external growth delay at early nymphal instars caused by high water temperatures in late summer and early autumn. Nymphs of S. manchuricus grew actively at 0.2°C during winter, which gave this species a competitive advantage over S. compacta and K. exilis whose growth was slow or absent in winter months. The semivoltine life cycle of K. exilis included a 9-month embryonic diapause and lasted about 4 years with periods of winter nymphal quiescence from December to February. Extended emergence and oviposition were recorded from the end of July to September. The population of K. exilis had a complex structure comprising 3 cohorts whose size ranges overlapped due to the extended flight period and recruitment of the new-generation nymphs in the spring of the next year. Nymphs of K. exilis grew fast during the warm period, when S. compacta and S. manchuricus were only appearing in the stream or underwent growth delay at relatively high water temperatures. The annual production of three species varied from 0.045 to 0.383 g/m2 dry mass, P/Bmean was 1.1–6.9; P/Bcoh = 6.2–9.1, and P/Byear = 4.1–9.4. Low values of P/Bmean and P/Byear were found in the semivoltine K. exilis, and high ones, in the univoltine S. compacta. The latter had a higher cohort-specific growth rate while its specific production was 0.026 day−1. The population of K. exilis had low values of the cohort-specific growth rate (0.013 day−1) and specific production (0.004 day−1). The highest production was observed in the population of S. manchuricus where nymphs grew at a specific rate of 0.023 day−1 while their mean density reached 59 ind./m2.

The complete mitochondrial genome and its remarkable secondary structure for a stonefly Acroneuria hainana Wu (Insecta: Plecoptera, Perlidae)

The Plecoptera (stoneflies) is a hemimetabolous order of insects, whose larvae are usually used as indicators for fresh water biomonitoring. Herein, we describe the complete mitochondrial (mt) genome of a stonefly species, namely Acroneuria hainana Wu belonging to the family Perlidae. This mt genome contains 13 PCGs, 22 tRNA-coding genes and 2 rRNA-coding genes that are conserved in most insect mt genomes, and it also has the identical gene order with the insect ancestral gene order. However, there are three special initiation codons of ND1, ND5 and COI in PCGs: TTG, GTG and CGA, coding for L, V and R, respectively. Additionally, the 899-bp control region, with 73.30% A+T content, has two long repeated sequences which are found at the 3′-end closing to the tRNAIle gene. Both of them can be folded into a stem-loop structure, whose adjacent upstream and downstream sequences can be also folded into stem-loop structures. It is presumed that the four special structures in series could be associated with the D-loop replication. It might be able to adjust the replication speed of two replicate directions.

The life cycle and production of three common stonefly species (Insecta, Plecoptera) in the Kedrovaya River (the south of Primorskii Territory)

The life cycles and production parameters of three common detritivorous stonefly species (Paracapnia khorensis, Amphinemura verrucosa, and Taenionema japonicum) from the Kedrovaya River at the Kedrovaya Pad Reserve (the south of Primorskii Territory, Russian Far East) were studied. Judging from the body size histograms, all these species are univoltine and have no winter diapause. Eggs develop directly; larval growth is slow, takes about 282–295 days, and proceeds even at low water temperatures in winter. The P. khorensis population is characterized by synchronous adult emergence and recruitment, as well as by slow continuous increase in larval linear size and body mass from June to March. Amphinemura verrucosa adults have a prolonged flight period from June to August, and their recruitment takes place for two months, till the end of November. Larvae grow during autumn and winter, but the most rapid growth is recorded in spring, when the water temperature rises, just before adult emergence in June and July. The development of T. japonicum is characterized by larval diapause in August and September when the water temperature is high. In October, larval growth is resumed and remains rapid through autumn and winter, up to adult emergence in May. The species studied had similar values of instantaneous growth rate (0.021–0.022 day−1), cohort production efficiency P/Bcohort (6.01–6.55), and annual efficiency of population production P/Bannual (7.78–8.11). More variable throughout the growing period was the mean biomass, and this accounted for the differences in secondary production values which ranged from 13.2 to 130.7 mg dry mass/m2.

A multi-trait approach for the identification and protection of European freshwater species that are potentially vulnerable to the impacts of climate change

We present a multi-trait approach to identify potentially vulnerable species of Ephemeroptera (mayflies), Plecoptera (stoneflies) and Trichoptera (caddisflies), collectively referred to as EPT, to the impacts of climate change (CC). The “climate change vulnerability score” (CCVS) is an aggregation of six autecological traits that are known to be associated with vulnerability to CC: endemism, micro-endemism, temperature preference, altitudinal preference, stream zonation preference, and life history. We assigned a vulnerability score (0 – invulnerable to 6 – highly vulnerable to climate change) to 1940 EPT species and discussed the applicability of the index at three spatial scales: (1) continental (Europe), (2) state (the German Federal State of North Rhine-Westphalia) and (3) a river basin (the Ruhr River). We identified 157 EPT species (ca. 8%) as highly vulnerable to climate change (CCVS≥4), including 95 species of caddisflies, 60 species of stoneflies and two species of mayflies. These are mostly found in France and Italy (52 species each), Spain and Slovenia (36 and 34, respectively), and Austria and Switzerland (30 species each), of which 95 are caddisflies, 60 stoneflies, and 2 mayflies. Using data collected in routine regional sampling we show that although no endemic EPTs were found in the German Federal State of North Rhine-Westphalia, eight species can still be identified as relatively vulnerable to CC (CCVS of 3). Almost all of these species are occurring in the ‘mountainous’ regions of the state (>200 m a.s.l.), the Sauerland and the Eifel. The upper reaches of the Ruhr catchment have been found to be relatively rich in vulnerable species, including several locally rare species. This index can assist conservationists to identify “hotspots” in terms of climate vulnerability and climate change refuge areas that can be considered for protection or the application of restoration measures at a local and regional scale. Nevertheless, not all species have complete autecological information, which hinders our ability to fully recognize the areas of priority. To further stabilize and enhance the applicability of this method, it is essential to fill these knowledge gaps in the future.

The complete mitochondrial genome of a stonefly species, Togoperla sp. (Plecoptera: Perlidae)

The complete mitochondrial (mt) genome of a stonefly species, Togoperla sp. (Plecoptera: Perlidae), was sequenced. The 15,723 bp long genome has the standard metazoan complement of 37 genes and an A+T-rich region, which is the same as the insect ancestral genome arrangement.