Chironomid pupal exuviae and larvae of two storage reservoirs in The Netherlands

For three years chironomid larvae were collected bimonthly with a grab sampler in two deep storage reservoirs in The Netherlands. Chironomid pupal exuviae were collected monthly, from April to November, with a handnet. Net samples yielded more taxa than grab samples. The taxa found in net samples originated from more habitats, than taxa found in grab samples. The relative contribution of Orthocladiinae was much larger in net samples. Based on the results obtained in the two reservoirs, some advantages and disadvantages of both sampling methods are discussed.     

Chironomid community structure deduced from larvae and pupal exuviae of a chalk stream

Species abundances of Chironomidae (Insecta: Diptera) have often been excluded from studies of benthic river communities because of difficulties associated with sampling and identifying larvae. Chironomid pupal exuviae are easier to collect and identify and could be used to determine community structure if shown to be representative of local larval assemblages. Larvae were sampled along a 20 m chain secured over mid-channel gravels, upstream of two collection points for pupal exuviae. Proportional taxa abundances of pupal exuviae and larvae sampled from 130 m of stream were directly compared by a ² test of independence and also separately fitted to four models of species abundance distribution. Observed proportions of taxa were not independent of the life stage sampled. The greatest discrepancies occurred with species of pupal exuviae that were absent as larvae from the gravel. The log series model provided the best fit with both pupal and larval data. Collections of pupal exuviae had greater species richness and evenness than samples of larvae. This was considered to be a consequence of sampling larvae from the gravel habitat alone.     

Water quality changes of the river Meuse assessed by chironomid pupal exuviae

The chironomid fauna of the river Meuse in Belgium and The Netherlands was studied in order to determine spatial water quality variations. Because the river Meuse serves as a source for drinking water production for about five million people in Belgium and The Netherlands, water quality is an essential factor to be studied. From 1984 till 1989 pupal exuviae were collected. Water quality was assessed by several indices and autecological information of the various taxa. With regard to pollution the general trend through the years was the same. The most upstream sampling site, in the Belgian Ardennes, was relatively unpolluted. As a result of untreated industrial and municipal wastewater discharges in the Liège region, water quality deteriorated dramatically at the Dutch border. At the most downstream sampling site, near the river mouth, a slight recovery was observed. Apart from organic and inorganic pollution other environmental factors such as flow velocity and the nature of the riverbed, also play a significant role in the occurrence of the organisms. It is concluded that protective measures need to be taken to restore the affected river ecosystem.The editors deeply regret to have to announce that Nicole Frantzen died in a train accident on 30 November 1992.     

Diel emergence patterns of drifting chironomid (Diptera) pupal exuviae in the Aude River (Eastern Pyrenees,France)

Two diel emergence patterns (April and October 1990) of chironomids from a hyporhithral site (4th–5th order) in the Aude River are described. In April, 62 species were collected with a mean hourly richness of 31 (±4). The Orthocladiinae (91%) largely dominated the emergence. The total number of exuviae captured by one drift-net varied from 1300 to 11600 per one hour period. There was a clear bimodal pattern with peaks post dawn (07.00–09.00 hrs) and around sunset (18.00 hrs).In October, the total number of exuviae captured was much lower and varied from 150 to 540 per one hour period with post dawn (06.00–08.00 hrs) and post sunset (18.00 hrs) peaks. Nevertheless, the total species richness (65) was higher as was the mean hourly richness (37±4). The Orthocladiinae (68%) dominated less because of increases in the Tanytarsini (15%) and Chiromini (14%). A diel rhythm appeared in autumn emergence. Most of the Orthocladiinae exuviae were collected during daylight hours, whereas most of the Chironomini and Tanytarsini were sampled during darkness. A majority of the exuviae with a length below 3 mm emerged during daylight, but most of the exuviae above 5 mm length emerged during darkness. The exuviae with an intermediate length (3–5 mm) had an amodal diel emergence pattern. In spring and autumn, the majority of species had a bimodal emergence pattern either by day (Orthocladiinae) or by night (Chironominae). But the smallest species had either an unimodal (Thiennemaniella vittata) or bimodal (Neozavrelia fuldensis) daytime emergence. The diel emergence patterns of 10 of the most frequently recorded species are presented.     

Analysis of chlordane-impacted streams using chironomid pupal exuviae (Diptera: Chironomidae)

A method of rapidly assessing streams and rivers using chironomid pupal exuviae was tested on a chlordane-impacted stream near St. Louis, Missouri. Various concentrations of chlordane were found in sediments of Grand Glaize Creek, most likely resulting from soil runoff around housing and business developments throughout the stream's course. Chironomid pupal exuviae and sediment samples were collected concurrently from Grand Glaize Creek on two separate occasions, once in 1988 and again in 1990. Cluster analysis of samples at sites, based on the percent abundances of taxa within habitats and by subfamilies, produced two distinct clusters; one grouping samples with lower chlordane concentrations and the other grouping samples with higher chlordane concentrations. Further analysis showed a trend towards lower percent abundances of taxa living in depositional and transitional/depositional zones (Chironominae and some Tanypodinae) at the higher chlordane site, while the lower chlordane sites had lower percent abundances of taxa within erosional and erosional/transitional zones (Orthocladiinae and some Tanypodinae). These findings support the hypothesis that taxa living in close association to fine organic sediments will be exposed to higher concentrations of chlordane in the stream and more negatively affected than taxa feeding and living in habitats removed from chlordane-bound sediments. Comparisons from the present study were made to related studies revealing similar patterns among the Chironomidae.     

A survey of the distribution of chironomidae (Diptera,Insecta) of the river rhine by sampling pupal exuviae

During the period from 1981-07-23 to 1981-08-10, the River Rhine, from the Alps to the North Sea, was sampled for chironomid pupal exuviae. Samples were also taken from the Grande Canal d'Alsace and from selected tributaries. In all 78 samples were taken, of which 9771 exuviae were examined and 135 taxa identified. A species list is given showing the distribution of taxa between the principal regions of the river (Appendix 1). Samples were also taken to show the within-sample and between-sample variability, and one pair of samples was used to demonstrate the difference between the two banks of the river below the junction with the Main. The Rhine above the Bodensee has a diverse fauna dominated by Orthocladiinae. The Hochrhein near the entry of the River Aare, and the River Aare itself, also have a diverse fauna, but include many sediment-dwelling Chironominae. Below Strasbourg the fauna changes markedly, losing most of its sediment-dwellers, and shows a great increase of pollution-tolerant species. The fauna generally becomes less diverse and of poorer quality throughout the Oberrhein sections, and faunal changes are demonstrated that are related to the inflow of the Rivers Neckar and Main, Samples from the Mittelrhein gorge and the Niederrhein are generally dominated by two species ofRheotanytarsus and show a progressive reduction of diversity coupled to an increase of pollution-tolerant species. The low proportions of sediment-dwellers in the Niederrhein and the Waal, suggests that the sediments are unsuitable for the support of a normal chironomid fauna.     

Trace DNA from insect skins: a comparison of five extraction protocols and direct PCR on chironomid pupal exuviae

Insect skins (exuviae) are of extracellular origin and shed during moulting. The skins do not contain cells or DNA themselves, but epithelial cells and other cell‐based structures might accidentally attach as they are shed. This source of trace DNA can be sufficient for PCR amplification and sequencing of target genes and aid in species identification through DNA barcoding or association of unknown life stages. Species identification is essential for biomonitoring programs, as species vary in sensitivities to environmental factors. However, it requires a DNA isolation protocol that optimizes the output of target DNA. Here, we compare the relative effectiveness of five different DNA extraction protocols and direct PCR in isolation of DNA from chironomid pupal exuviae. Chironomidae (Diptera) is a species‐rich group of aquatic macroinvertebrates widely distributed in freshwater environments and considered a valuable bioindicator of water quality. Genomic DNA was extracted from 61.2% of 570 sampled pupal exuviae. There were significant differences in the methods with regard to cost, handling time, DNA quantity, PCR success, sequence success and the ability to sequence target taxa. The NucleoSpin^® Tissue XS Kit, DNeasy^® Blood and Tissue kit, and QuickExtract^? DNA Extraction Solution provided the best results in isolating DNA from single pupal exuviae. Direct PCR and DTAB/CTAB methods gave poor results. While the observed differences in DNA isolation methods on trace DNA will be relevant to research that focuses on aquatic macroinvertebrate ecology, taxonomy and systematics, they should also be of interest for studies using environmental barcoding and metabarcoding of aquatic environments.     

Spatial and temporal distribution patterns of drifting pupal exuviae of Chironomidae (Diptera) in streams of tropical northern Australia

1. Periodic collecting of floating cast pupal cuticles of chironomids (exuviae) in two tropical northern Australian streams demonstrates (i) spatial heterogeneity in species composition across a wide stream, (ii) temporal heterogeneity in the maximum abundance of each species, and that (iii) species accumulate as a function of sample size and duration of sampling, 2. Spatial heterogeneity is ascribed to variation in larval microhabitat across the wide stream, combined with short exuvial drift duration and restricted upstream mixing. 3. Temporal heterogeneity is ascribed to diel periodicity in adult emergence and, as with spatial heterogeneity, to the short floating life. 4. The consequences of spatial and temporal variation for the sampling of exuvial drift are discussed in relation to the objectives of particular programmes. Thus, if the objective is assessment of chironomid species composition for inventory purposes such as faunistics or conservation, the large sample sizes attained by 24-h sampling are necessary and appropriate. However, for rapid assessment that requires comparable samples at different sites, species numbers may be optimized by temporally and spatially restricted sampling of the maximal emergence period, which in this study is at dusk, or by subsampling from a 24-h sample.     

Downstream drift of the larvae of Chironomidae (Diptera) in the River Chew,S.W. England

The drift of larval Chironomidae (Diptera) was investigated at two sites on the River Chew, using a pump filtration system with 50 µm mesh-aperture aerial nets, situated on the river bank.Chironomid larvae were found to drift in far greater numbers than previously reported, even in slow-flowing water. First and second instar larvae dominated the drift.Variation in drifting behaviour between taxa was observed, where Orthocladiinae drifted in all instars and Chironominae predominantly as first and second instars. Proportionally different rates of drift were observed between these taxa from the benthos at the two sites.It is suggested that the majority of chironomid drift represents an active dispersal and colonisation mechanism by which population redistribution and habitat selection occurs.     

Communities of Chironomid larvae in the Velika Morava river

The following communities of Chironomid larvae were found: psamorheophile, psamophile, pelorheophile, pelophile and phytorheophile. They differ according to both their coenotical composition and the number of organisms, and show a particular distribution along the river course in correlation with the type of substrate. The most widespread are the psamorheophile communities and the most restricted the phytorheophile ones.     

The Application of Chironomid Pupal Exuvial Technique (CPET) for Ecological Analysis in a Neotropical Large River System

This field ecological study, based on the chironomid pupal exuvial technique (CPET), is new for the Paraná River and proposes an efficient tool to be used in future ecological approaches and biomonitoring. Drifting of pupal exuviae in a river-floodplain system of the Middle Paraná River floodplain was represented by 34 Chironomidae taxa, being the characteristic association obtained from the CPET: Lopescladius, Onconeura, Paralauterborniella, Polypedilum, and Harnischia complex. Diversity, richness, dominance, total density, and density of dominant taxa were different between the longitudinal and lateral dimensions but not between hydrologic phases, with a greater diversity and richness in the main channel of the river and higher density and dominance in floodplain habitats. The species turnover is the dominant process in structuring studied assemblages in spatial and temporal analysis, increasing in the floodplain habitats and in low-water phase. The results obtained showed that drifting exuviae in the longitudinal axis were coming from different assemblages and environments of a wider area (regional), while exuviae recorded in the connections of the floodplain environments in the lateral dimension could reflect the local assemblages. We demonstrated the ecological value of CPET studies to interpret the attributes of Chironomidae assemblage in river-floodplain systems of large rivers in an integrated way.     

Cellular behavior in the developing Drosophila pupal retina

Correct patterning of cells within an epithelium is key to establishing their normal function. However, the precise mechanisms by which individual cells arrive at their final developmental niche remains poorly understood. We developed an optimized system for imaging the developing Drosophila retina, an ideal tissue for the study of cell positioning. Using this technique, we characterized the cellular dynamics of developing wild-type pupal retinas. We also analyzed two mutants affecting eye patterning and demonstrate that cells mutant for Notch or Roughest signaling were aberrantly dynamic in their cell movements. Finally, we establish a role for the adherens junction regulator P120-Catenin in retinal patterning through its regulation of normal adherens junction integrity. Our results indicate a requirement for P120-Catenin in the developing retina, the first reported developmental function of this protein in the epithelia of lower metazoa. Based upon our live visualization of the P120-Catenin mutant as well as genetic data, we conclude that P120-Catenin is acting to stabilize E-cadherin and adherens junction integrity during eye development.     

Chironomid microdistribution in gravel of an English chalk river

1. The study aimed to determine the relative importance of certain environmental variables to the distribution of instars of chironomid species in mid-stream gravels of the River Pang. 2. Core samples of gravel substrata were collected over 1 year. Water depth, current, particle size distribution, substratum porosity and detritus were recorded for each sample. The four principal components from a linear ordination of the environmental variables were used to constrain the ordination of chironomid data using the program CANOCO. 3. Microdistribution was principally related to a temporal gradient of water depth, current and substratum heterogeneity. There was significant spatial separation of instars along a gradient of fine detritus, silt and substratum porosity. The distributions of early instars of three taxa were also significantly related to particle size. There was a significant relationship between the size distribution of larvae and the spatial distribution of fine sediments and detritus. This relationship was strengthened when restricted to larvae classified as scrapers.     

Chironomid midge sensitization in sewage workers: case study

Non‐biting chironomid midges (Diptera: Chironomidae) may cause sensitization and allergic reactions in humans and have recently been identified as a potential health problem in Swedish municipal sewage treatment plants. To investigate, on a pilot scale, the allergenic potential of chironomids in sewage workers, all workers (n = 8) at a sewage treatment plant and local controls (n = 16) completed a symptom questionnaire, underwent measurement of the fraction of nitric oxide in exhaled air, spirometry, and provided serum samples for the determination of atopy status and the prevalence of specific immunoglobulin E (IgE) antibodies against Chironomus thummi (Chi t) using a commercial fluorescence enzyme immunoassay (FEIA). Three sewage workers (38%) but no controls (0%) were FEIA positive for C. thummi‐specific IgE antibodies (P < 0.05). No other health‐related findings were significantly different between the groups. The study suggested that occupational exposure to Chironomids may cause sensitization with circulating IgE‐antibodies in sewage workers.     

Epidermal feet in pupal segment morphogenesis

Epidermal cells in insect integumental epithelia develop branched cytoskeletal extensions or feet at their base that are similar in appearance to the processes put out by cells in tissue culture. We have developed a procedure to show the feet that gives an effect as if thousands of cells randomly arranged in the epithelium had each been injected with a lead salt visualized as black lead sulphide. The procedure depends upon the fact that after brief glutaraldehyde fixation, tannic acid only penetrates some cells where it mordants lead ions and binds osmium. Individual cells visualized in this manner show their outlines as if they are separate in a tissue culture although they are part of a closely packed epithelium. The feet are metamorphic structures formed after pupal commitment and are necessary for metamorphic changes in segment shape. In Calpodes larvae the feet are orientated axially in the direction of the segmentally repeating gradient and may extend for several cell diameters. They extend under the influence of low titres of 20-hydroxyecdysone such as those occurring in the intermoult. When stimulated by high titres like those in pre-pupae, the feet contract at the same time as the segments shorten to pupal proportions. We believe that cell processes like the epidermal feet are ubiquitous but that they have often been overlooked because of the difficulty of demonstrating the outlines of single cells that are united in epithelia.     

The composition of the free fatty acids from Dendrolimus pini exuviae

The pine moth Dendrolimus pini effectively resists many insecticides, but it can be controlled by the use of bioinsecticides such as entomopathogenic fungi. In the use of microbial agents for the biocontrol of D. pini, it is important to identify the cuticular lipids of this pest if we are to understand the factors responsible for the preferential adhesion or selective repulsion of entomopathogenic fungi that are potentially useful in biocontrol. In this work the qualitative and quantitative analyses of free fatty acids in two exuviae extracts (petroleum ether and dichloromethane) and two developmental stages (larval-larval and larval-pupal molts) were studied. The free fatty acid composition of the epicuticular lipids from exuviae of D. pini was characterized chemically using gas chromatography (GC) and gas chromatography-electron impact mass spectrometry (GC-MS). Structural analyses of the dichloromethane extracts from larval-larval exuviae (LLE) and larval-pupal exuviae (LPE) revealed that the carbon numbers for the major acid moieties ranged from C_8:0 to C_34:0. Only C_23:0 was not identified in the LPE extract. The relative contents of fatty acids in the extracts varied from trace amounts to 34%. The fatty acids extracted by dichloromethane were essentially the same as those in the petroleum ether extract. We also identified dehydroabietic acid in the exuviae of D. pini. The respective quantities of dehydroabietic acid obtained from D. pini LLE and LPE were 1763 ± 103 μg/g exuviae and 11521 ± 1198 μg/g of exuviae.     

Hormonal control of lutein incorporation into pupal cuticle of the butterfly Inachis and the pupal melanization reducing factor

Abstract. . Morphological colour adaptation of pupae of the butterfly Inachis io L. (Lepidoptera: Nymphalidae) is controlled by a factor which reduces cuticular melanization (Biickmann & Maisch, 1987). This so-called pupal melanization reducing factor (PMRF) is located throughout the entire central nervous system of prepupae (Stamecker et al. , 1994).
Extracts of abdominal ganglia also stimulated dose-dependently lutein incorporation into pupal cuticle. In the bioassay higher doses were required to increase cuticular lutein content than to reduce melanization. Ligatures during the prepupal stage demonstrated two different critical periods for these pigmentation effects: an early one for melanization reduction and a late one for lutein incorporation.
An initial chromatographic purification yielded only two adjacent fractions which contained both the PMRF and the stimulation of lutein incorporation activity. Therefore it is assumed that only one hormone with a dual function may be responsible for pupal pigmentation.
Lutein content was found in gut, fat body, epidermis and haemolymph of I.io. Lutein incorporation into cuticle occurred within 1.5 days of the pupal moult when the cuticle was not yet fully sclerotized. Lutein content is significantly higher in cuticle of yellow pupae than of black ones.     

The emergence of order in the Drosophila pupal retina

During pupation, long-range order is imposed on the autonomously developing ommatidia which compose the Drosophila eye. To accomplish this, eight additional cell types arise: the primary, secondary, and tertiary pigment cells, and the four cells that form the bristle. These cells form an interweaving lattice between ommatidia. The lattice is refined when excess cells are removed to bring neighboring ommatidia into register. Recent evidence suggests that in larval development, local contacts direct cell fate. The same appears to be true during pupal development: the contacts a cell makes predict the cell type it will become. Cells which contact the anterior or posterior cone cells in an ommatidium invariably become primary pigment cells. Cells which contact primary pigment cells from different ommatidia become secondary and tertiary pigment cells. Bristle development is in several ways distinct from ommatidial development. The four cells of each bristle group appear to be immediate descendents of a single founder cell. During their early differentiation, they do not make stereotyped contacts with surrounding ommatidial cells, but do make particular contacts within the bristle group. And unlike the surrounding ommatidia, differentiation of the bristles radiates from the center of the eye to the edges. As cells are removed during two stages of programmed cell death, the bristles are brought into their final position. When all cells in the lattice have achieved their final position, a second stage of retinal development begins as structures specific to each cell type are produced. This paper follows these various stages of pupal development, and suggests how local cell-cell contacts may produce the cells needed for a functional retina.     

Operations on the pupal wing ofDrosophila melanogaster

Journal of Genetics -