Response of Chironomids (Diptera, Chironomidae) to Environmental Factors at Different Spatial Scales.

Factors responsible for species distribution of benthic macroinvertebrates, including responses at different spatial scales, have been previously investigated. The aim of the present research was to review the most relevant factors explaining chironomid species distribution focusing on factors operating at different spatial scales, such as latitude, longitude, altitude, substrate, salinity, water temperature, current velocity, conductivity, acidity, dissolved oxygen, nutrient content etc. acting at regional levels and at a large or small water basin level. Data including chironomid species abundances from different lentic and lotic waters in Italy and other surrounding countries were analyzed using partial canonical correspondence analysis (pCCA) and multiple discriminant analysis (DISCR). Spatial analyses, including univariate Moran's I correlograms, multivariate Mantel correlograms and Moran's eigenvector maps (MEMs), were thereafter carried out. The results showed that habitat type, including different types of lotic waters (i.e., kryal, crenal, rhithral, potamal) and different lake types (i.e., littoral, sublittoral, profundal zones), is the most significant factor separating chironomid assemblages, while spatial factors act only as indirect influencers.

Response of Chironomidae (Diptera) to DDT, Mercury, and Arsenic Legacy Pollution in Sediments of the Toce River (Northern Italy).

The Toce River (Northern Italy) is characterized by legacy contamination of dichloro-diphenyl-trichloroethane (DDT), mercury, and arsenic deriving from an industrial plant active between ca. 1915 and 1996. Chironomidae taxa assemblages and sediments were collected in 2014 and 2019 upstream and downstream of the industrial area to analyze species responses to toxic substances in a river stretch with relatively uniform natural (i.e., hydro-morphological) characteristics. A total of 32 chironomid taxa were identified. Sediment concentrations reached levels potentially toxic for benthic invertebrates: 15.7 µg kg-1 1% organic carbon for DDT, 197 µg kg-1 dry weight (d.w.) for Hg, and 55.7 mg kg-1 d.w. for As. Canonical Correspondence Analysis (CCA) revealed a predominant seasonal gradient, followed by an upstream-downstream gradient. Partial CCA indicated that 5.2% of the total variation was associated with sediment contamination. Self-Organizing Maps (SOMs) were used to represent species responses to toxicants. Most species appeared to be tolerant, e.g., Chironomus riparius, Micropsectra atrofasciata, Conchapelopia pallidula, and Polypedilum spp. Sensitivity to contaminants was observed in only a few species: Diamesa spp., Sympotthastia spinifera, and Prodiamesa olivacea to DDT; Potthastia longimanus to Hg; Odontomesa fulva and Microtendipes pedellus to As. The chironomid community was characterized in presence of contamination levels commonly observed in freshwater ecosystems.

Response of Chironomids to Key Environmental Factors: Perspective for Biomonitoring.

Chironomids are the species-richest family among macroinvertebrates and are often used as indicators of ecological conditions in inland waters. High taxonomic expertise is needed for identification and new species are still being described even in the well-known West Palearctic region. Our Microsoft Access relational database comprises data on Chironomid species collected in rivers and lakes in Italy and some other European countries over a period of about 50 years, often associated with physical-chemical data, but in some cases, only data on Chironomids are available with no associated environmental data. The aim of the present paper was to propose the calculation of ecological traits of Chironomid species as a tool to derive information on water quality, when only data on Chironomid species composition are present, while environmental data are lacking. Traits summarizing the species' response to environmental variables were evaluated, with emphasis on natural and man-influenced factors: current velocity, water temperature, conductivity, dissolved oxygen, and nutrients. Traits calculations were carried out in the R environment using a subset of our data, including both environmental data and Chironomid abundances. The relations between sites, Chironomid, species and traits were evaluated using correspondence analysis and other multivariate methods. The response of species showed an interaction among different factors, with the possibility of ordering species along a single environmental gradient, extending from cold running waters to warm standing waters, with few exceptions.

Corrections and Additions to Descriptions of Some Species of the Subgenus Orthocladius s. str. (Diptera, Chironomidae, Orthocladiinae).

The larvae of some species of the subgenus Orthocladius s. str. (Diptera, Chironomidae) are here described for the first time with corrections and additions to the descriptions of adult males and pupal exuviae. The identification of larvae is generally not possible without association with pupal exuviae and/or adult males, so the descriptions here are based only on reared material or on pupae with the associated larval exuviae. Usually, Chironomidae larvae can be separated on the basis of morphometric characters, the most discriminant ones are: (1) the ratio between the width of median tooth of mentum (Dm) and the width of the first lateral tooth (Dl) = mental ratio (DmDl), (2) the ratio between the length of the first antennal segment (A1) and the combined length of segments 2-5 (A2-5) = antennal ratio (AR). The shape of mandible, maxilla, and other body parts are almost identical in all the species considered in this study. The larva of Orthocladius (Symposiocladius) lignicola is very characteristic and can be separated by the shape of mentum and the larvae of all the known species of Symposiocladius are characterized by the presence of large Lauterborn organs on antennae and of tufts of setae on abdominal segments. The larvae of Orthocladius (Orthocladius) oblidens and Orthocladius (Orthocladius) rhyacobius can be distinguished from other species basing on their large Dm and to each other by AR. A principal component analysis was carried out using 5 characters: (1) Dm, (2) Dl, (3) length of A1, (4) width of A1 (A1W), (5) combined length of segments 2-5 (A2-5). The most discriminant characters were Dm and A1, confirming that DmDl and AR can be used to separate species at larval stage, but the large superposition of morphometric characters in different species confirms that association with pupal exuviae is in any case needed to identify larvae. In future perspective, the development of reference DNA barcodes from specimens identified by specialists is recommended since possibly the best tool for larvae identification, but association of barcodes with morphotypes is in any case fundamental.

Twenty years of Dipterology through the pages of Zootaxa.

We present a summary and analysis of the Diptera-related information published in Zootaxa from 2001 to 2020, with a focus on taxonomic papers. Altogether, 2,527 papers on Diptera were published, including 2,032 taxonomic papers and 1,931 papers containing new nomenclatural acts, equivalent to 22% of all publications with new nomenclatural acts for Diptera. The new nomenclatural acts include 7,431 new species, 277 new genera, 2,003 new synonymies, and 1,617 new combinations. A breakdown by family of new taxa and new replacement names proposed in the journal during the last two decades is provided, together with a comparison of Zootaxa's output to that of all other taxonomic publications on Diptera. Our results show that the journal has contributed to 20% of all biodiversity discovery in this megadiverse insect order over the last 20 years, and to about 31% in the last decade.

The Atopy Index Inventory: A Brief and Simple Tool to Identify Atopic Patients.

INTRODUCTION: Atopy and ear, nose and throat (ENT) diseases are frequently associated; however, no clinical tool has been proposed so far to discriminate which patients could be atopic and therefore deserving of a further immunoallergological evaluation. OBJECTIVE: The aim of this study was to assess and validate a set of dichotomous responses suitable for predicting the presence of atopy in adult patients. METHODS: An 11-item questionnaire, i.e., the Atopy Index Inventory (AII), comprised of 4 questions regarding the clinical history for allergic disease and 7 questions evaluating the presence of the most frequent clinical signs affecting allergic patients, was developed and administered to 226 adult subjects (124 atopic subjects and 102 healthy, not atopic subjects). The atopic condition was proven by an immunoallergological evaluation according to the diagnostic criteria of the EAACI guidelines. Internal consistency and clinical validity were tested. RESULTS: In healthy subjects, the first 4 variables of the AII returned a 100% correct response (all answered "no") and were defined as "decisive" responses. In the logistic regression analysis, when decisive items were negative, the atopic condition was confirmed when answering "yes" to at least 3 "probability" items (cut-off = 2.69). The difference in AII scores between allergic and healthy group was significant using the Mann-Whitney U test (p < 0.0001). The sensitivity and specificity of the AII were 0.97 and 0.91, respectively, with a true predictive value of 0.92 and a false predictive value of 0.97. The ROC curve showed an area of 0.94, with an OR of 0.88 (95% CI 0.87-0.97, p = 0.0001). The internal consistency as determined by the Cronbach α coefficient was 0.88. CONCLUSION: The AII has been proven to be a brief, simple and sufficiently accurate tool for screening ENT patients in search of atopic individuals and to allow their clinical management.

The species of the genus Diamesa (Diptera, Chironomidae) known to occur in Italian Alps and Apennines.

Some rare species from Italian Alps, belonging to the genus Diamesa Meigen, 1835 (Diptera, Chironomidae) are here redescribed as adult males, because only old, incomplete descriptions are available for these taxa. Terminology of male genitalia is reviewed, diagnostic features are illustrated in detail, and notes on biology and geographical distribution of the examined species are provided. An identification key to the known adult males is presented.

Correction: Integrated Taxonomy and DNA Barcoding of Alpine Midges (Diptera: Chironomidae).

[This corrects the article DOI: 10.1371/journal.pone.0149673.].

Transcriptional profiling induced by pesticides employed in organic agriculture in a wild population of Chironomus riparius under laboratory conditions.

Copper (Cu) and azadirachtin (AZA-A+B) are pesticides allowed in organic agriculture whose environmental risk and toxicity for aquatic wildlife is only partially known. Reverse Transcription Polymerase Chain Reaction was used to assess the molecular effect of acute and short-term exposure (3, 24h) of Cu (0.01, 0.05, 1, 10, 25mgl(-1)) and AZA-A+B (0.2, 0.3, 0.4, 0.5, 1mgl(-1)) on the expression of five candidate genes (hsp70, hsc70, hsp40, hsp10 and cyP450) in a non-target species, Chironomus riparius. Fourth-instar larvae were collected from a mountain stream polluted by agricultural land run-off. All genes were responsive to both pesticides but each gene had a specific response to the different experimental concentrations and exposure times. A few similarities in transcriptional profiling were observed, such as a linear concentration-dependent response of hsp70 after 24h of exposure (at ≥1mgl(-1) of Cu and ≥0.2mgl(-1) of AZA-A+B) and an up-regulation regardless of the concentration of hsc70 after 24h of exposure (at ≥0mgl(-1) of Cu and ≥0.2mgl(-1) of AZA-A+B and the up-regulation of hsp70 after 3h of exposure at ~LC50 (Cu-LC50=26.1±2.5mgl(-1), AZA-A+B-LC50=1.1±0.2mgl(-1)). According to the results, hsp40, hsp10 and cyP450 may be defined as pesticide-dependent (i.e., hsp40 and hsp10 seemed to responded mainly to AZA-A+B and cyP450 to Cu), while hsc70 as time-dependent regardless of the pesticide (i.e., hsc70 responded only after 24h of treatment with Cu and AZA-A+B). This study gives new insights on the potential role of the C. riparius's hsps and cyP450 genes as sensitive biomarkers for freshwater monitoring.

Integrated Taxonomy and DNA Barcoding of Alpine Midges (Diptera: Chironomidae).

Rapid and efficient DNA-based tools are recommended for the evaluation of the insect biodiversity of high-altitude streams. In the present study, focused principally on larvae of the genus Diamesa Meigen 1835 (Diptera: Chironomidae), the congruence between morphological/molecular delimitation of species as well as performances in taxonomic assignments were evaluated. A fragment of the mitochondrial cox1 gene was obtained from 112 larvae, pupae and adults (Diamesinae, Orthocladiinae and Tanypodinae) that were collected in different mountain regions of the Alps and Apennines. On the basis of morphological characters 102 specimens were attributed to 16 species, and the remaining ten specimens were identified to the genus level. Molecular species delimitation was performed using: i) distance-based Automatic Barcode Gap Discovery (ABGD), with no a priori assumptions on species identification; and ii) coalescent tree-based approaches as the Generalized Mixed Yule Coalescent model, its Bayesian implementation and Bayesian Poisson Tree Processes. The ABGD analysis, estimating an optimal intra/interspecific nucleotide distance threshold of 0.7%-1.4%, identified 23 putative species; the tree-based approaches, identified between 25-26 entities, provided nearly identical results. All species belonging to zernyi, steinboecki, latitarsis, bertrami, dampfi and incallida groups, as well as outgroup species, are recovered as separate entities, perfectly matching the identified morphospecies. In contrast, within the cinerella group, cases of discrepancy arose: i) the two morphologically separate species D. cinerella and D. tonsa are neither monophyletic nor diagnosable exhibiting low values of between-taxa nucleotide mean divergence (0.94%); ii) few cases of larvae morphological misidentification were observed. Head capsule color is confirmed to be a valid character able to discriminate larvae of D. zernyi, D. tonsa and D. cinerella, but it is here better defined as a color gradient between the setae submenti and genal setae. DNA barcodes performances were high: average accuracy was ~89% and precision of ~99%. On the basis of the present data, we can thus conclude that molecular identification represents a promising tool that could be effectively adopted in evaluating biodiversity of high-altitude streams.

Finite-size scaling as a way to probe near-criticality in natural swarms.

Collective behavior in biological systems is often accompanied by strong correlations. The question has therefore arisen of whether correlation is amplified by the vicinity to some critical point in the parameters space. Biological systems, though, are typically quite far from the thermodynamic limit, so that the value of the control parameter at which correlation and susceptibility peak depend on size. Hence, a system would need to readjust its control parameter according to its size in order to be maximally correlated. This readjustment, though, has never been observed experimentally. By gathering three-dimensional data on swarms of midges in the field we find that swarms tune their control parameter and size so as to maintain a scaling behavior of the correlation function. As a consequence, correlation length and susceptibility scale with the system's size and swarms exhibit a near-maximal degree of correlation at all sizes.

Collective behaviour without collective order in wild swarms of midges.

Collective behaviour is a widespread phenomenon in biology, cutting through a huge span of scales, from cell colonies up to bird flocks and fish schools. The most prominent trait of collective behaviour is the emergence of global order: individuals synchronize their states, giving the stunning impression that the group behaves as one. In many biological systems, though, it is unclear whether global order is present. A paradigmatic case is that of insect swarms, whose erratic movements seem to suggest that group formation is a mere epiphenomenon of the independent interaction of each individual with an external landmark. In these cases, whether or not the group behaves truly collectively is debated. Here, we experimentally study swarms of midges in the field and measure how much the change of direction of one midge affects that of other individuals. We discover that, despite the lack of collective order, swarms display very strong correlations, totally incompatible with models of non-interacting particles. We find that correlation increases sharply with the swarm's density, indicating that the interaction between midges is based on a metric perception mechanism. By means of numerical simulations we demonstrate that such growing correlation is typical of a system close to an ordering transition. Our findings suggest that correlation, rather than order, is the true hallmark of collective behaviour in biological systems.

Predicting PCB concentrations in cow milk: validation of a fugacity model in high-mountain pasture conditions.

A fugacity model reported in the literature was applied to a high-altitude pasture in the Italian Alps. The model takes into account three compartments (digestive tract, blood and fat tissues) in unsteady-state conditions using food as the contamination source. Disregarding biotransformation inside cow tissues, the predicted concentrations of 14 polychlorinated biphenyls (PCBs) in milk were in good agreement with the observed data, especially for congeners known for their resistance to biotransformation (e.g., CB-138 and 153). In contrast, the predicted concentrations were clearly overestimated for congeners with high biotransformation susceptibilities. Therefore data measured in milk and faeces were used to calculate the first-order-biotransformation rate constants in dairy cows. The PCB absorption efficiency observed for pasture conditions was lower than that observed in the cowshed. The final version of the model included biotransformation and observed PCB absorption and was able to predict PCB concentrations in cow milk with mean differences between the predicted and measured data below ± 20% for most congeners.

Landscape-stream interactions and habitat conservation for amphibians.

Semiaquatic organisms depend on the features of both water bodies and landscapes; the interplay between terrestrial and aquatic systems might influence the semiaquatic communities, determining the scale at which management would be more effective. However, the consequences of such interplay are not frequently quantified, particularly at the community level. We analyzed the distribution of amphibians to evaluate whether the influence of landscape features on freshwater ecosystems can have indirect consequences at both the species and community level. We surveyed 74 streams in northern Italy to obtain data on breeding amphibians, water, and microhabitat features; we also measured features of surrounding landscapes. We used an information-theoretic approach and structural equation models to compare hypotheses on causal relationships between species distribution and variables measured at multiple levels. We also used a constrained redundancy analyses to evaluate causal relationships between multivariate descriptors of habitat features and community composition. Distribution of Salamandra salamandra was related to landscape, hydrological, and water characteristics: salamanders were more frequent in permanent streams with low phosphate concentration within natural landscapes. Water characteristics were dependent on landscape: streams in natural landscapes had less phosphates. Landscape influenced the salamander both directly and indirectly through its influence on phosphates. Community structure was determined by both landscape and water characteristics. Several species were associated with natural landscapes, and with particular water characteristics. Landscape explained a significant proportion of variability of water characteristics; therefore it probably had indirect effects on community. Upland environments play key roles for amphibians, for example, as the habitat of adults, but upland environments also have indirect effects on the aquatic life stages, mediated through their influence on water characteristics. Synergistic effects can magnify the negative consequences of landscape alteration on amphibians; landscape management can be particularly effective, as it can also improve wetland features.

Increased alanine concentration is associated with exposure to fenitrothion but not carbamates in Chironomus riparius larvae.

Chironomus riparius Meigen were exposed to three different insecticides, the organophosphorous fenitrothion and the carbamates carbaryl and carbofuran (0, 1, 10, and 100 microg/L) for 24h as fourth-instar larvae. Acetylcholinesterase (AChE), naphtylacetate esterase (NAE), p-nitrophenylacetate esterase (PNPAE), glutathione-S-transferase (GST), and a number of metabolites (alanine, pyruvate, lactate, trehalose, aspartate, oxalacetate) were measured to determine which was the most valuable biochemical biomarker of exposure. AChE activity was significantly reduced by all three insecticides, PNPAE by fenitrothion, carbofuran and carbaryl, whereas NAE activity was stimulated by carbaryl and unaffected by fenitrothion and carbofuran. Metabolites analysis revealed a strong accumulation of alanine in larvae exposed to fenitrothion, but not in larvae exposed to carbamates. This accumulation was accompanied by a significant increase of lactate and a significant decrease of pyruvate and trehalose. No variations were observed with carbofuran and carbaryl. No change of aspartate concentration was detected. We conclude that the association of alanine accumulation with a significant inhibition of AChE activity can be used as a valuable biochemical biomarker of exposure.