Variation and divergence of multilocus genome markers in the species of the genus Chironomus (Diptera, Chironomidae)

Variation and divergence patterns of the multilocus genome markers in twelve Chironomus species belonging to the plumosus and piger sibling-species groups were examined by use of polymerase chain reaction with random primers (RAPD method). The chironomid species showed high levels of the RAPD markers polymorphism. The genetic distances (GD) were determined between the species within the group of closely related species, as well as between the species from different groups. The estimates obtained characterized the divergence levels between the sibling species (GD = 0.739) and morphologically distinct species (GD = 0.935). Comparison of the variation and divergence levels of the RAPD markers with those for the other genome markers, namely, the enzyme-coding genes and chromosomes (gene linkage groups) have demonstrated different divergence rates for different genome components during speciation of Chironimids.     

A review of the genus Chironomus (Diptera,Chironomidae)

Analysis of the banding pattern of the salivary gland chromosomes of Chironomus tepperi indicates that, despite a somewhat modified male hypopygium, the relationships of this species are close to the other Australian species of the genus, particularly to Ch. oppositus. No inversion polymorphism has been found in Ch. tepperi and this, together with the relatively high chiasma frequency as measured at metaphase I, would appear to be an adaptation to provide genetic variability necessary for its colonizing ability.     

Development and localization of microsatellite markers for the sibling species Chironomus riparius and Chironomus piger (Diptera: Chironomidae)

Five variable microsatellite loci are reported for the nonbiting midge species Chironomus riparius and Chironomus piger. All loci show considerable intraspecific variation and species‐specific alleles, which allow to discriminate among the two closely related species and their interspecific hybrids, and to estimate genetic diversity within and between populations. Additionally, the loci were localized on C. riparius polytene chromosomes to verify their single copy status and investigate possible chromosomal linkage. The described markers are used in different studies with regard to population and ecological genetics and evolutionary ecotoxicology of Chironomus.     

Polymorphism and divergence of multilocus DNA markers in sibling species Chironomus riparius Meigen and Chironomus piger Strenzke (Diptera, Chironomidae)

Intra- and interspecific variation and divergence of multilocus markers for genomic DNA of the sibling species from the thimmi group, Chironomus riparius and C. piger, were studied by PCR with arbitrary primers (RAPD). A high level of RAPD polymorphism was determined in both laboratory and natural populations of these species. The genetic distances were estimated between the C. riparius populations and between the sibling species C. riparius and C. piger. The genetic distance between C. riparius and C. piger was 4 to 5 times higher than that between the C. riparius populations. A comparison of the variation and divergence for the RAPD markers with those for other genomic markers--enzyme-coding genes and chromosomes (linked gene groups)--showed that different components of the genome differed in their contribution to the genome divergence.     

Karyotype structure in species of Propsilocerus genus (Diptera, Chironomidae)

Karyotype structure and polytene chromosome banding patterns were studied in two Orthocladiinae siblings--Propsilocerus akamusi (China) and Propsilocerus jacuticus (Russia). Both species have haploid number of chromosome typical for Orthocladiinae (n = 3). An unusual structure of centromeric regions was observed in all three chromosomes of karyotypes in both species. Photomaps of polytene chromosomes are presented. A comparison of karyotypes of P. akamusi and Propsilocerus jacuticus revealed a high level of homology in their banding sequences, however, the presence of fixed paracentric inversions in chromosomal arms IR, IIR, IIIR of Propsilocerus jacuticus has shown a clear-cut phylogenetic divergence. No chromosomal polymorphism was found in both species.     

DNA barcoding supports reclassification of Japanese Chironomus species (Diptera: Chironomidae)

Non‐biting midges (Diptera: Chironomidae) adapt to species‐specific environmental conditions and hence are promising bioindicators for aquatic and ecotoxicological monitoring. Although their utility for these purposes was historically limited by difficulties in their morphological identification, DNA barcoding offers a possible solution. Here, eight Japanese species of the genus Chironomus, which is characterized by its worldwide distribution and abundance among Chironomidae, were subjected to DNA barcoding using cytochromec oxidase subunit I (COI). To examine whether this DNA barcode is a useful indicator for Japanese species of Chironomus, we calculated genetic distances within and between the COI sequences of Chironomus species both from this study and worldwide and constructed phylogenetic trees. Based on 415 bp COI sequences and the Kimura two‐parameter model, the average genetic distances within 37 species and between 72 species were 2.6% and 17.2%, respectively. Although the ranges of genetic distances within and between species overlapped from 0.8% to 17.3%, 99.7% of average genetic distances between species were >3.0%. Some of this overlap is attributable to distances within species that were “too large” as well as those between species that were “too small”. Of eight Japanese species examined, two showed genetic distances between species that were below a 3.0% threshold, and four had distances within species that were greater than 3.0%. These results suggest a possible reclassification of these species and the need for further sampling to unveil biogeographic variations among different countries and regions.     

Cytogenetic characteristics of species of the Chironomus plumosus group (Chironomidae, Diptera) in Finland

The cytogenetic characteristics of species of the Chironomus plumosus group in Finland were examined. The species included C. balatonicus Devai, Wülker, Scholl, C. entis Shobanov, C. plumosus L., C. muratensis Ryser, Scholl, Wülker, and two karyotypes of unknown species. All belonged to the thummi complex having 2n = 8 chromosomes, with arm combinations of AB, CD, EF, G. In C. balatonicus arms A to G the band sequences corresponded with those of other populations. A new band sequence was found in arm B. In C. entis an arm A had a band sequence similar to those in C. agilis. A large pericentric inversion was observed in chromosome AB. The arms B to G had band sequences typical for C. entis. The chromosome arms A and B in C. plumosus from Lake Marsjon had band sequences corresponding to those of C. agilis and C. entis, respectively. C. plumosus from Helsinki may be a more divergent population with a large amount of centromere heterochromatin. C. muratensis was not distinguishable by band sequences from those of other Palaearctic populations. Two new karyotypes similar to those of species of the plumosus group have been described.     

A new species of Chironomus Meigen, 1803 (Diptera,Chironomidae) from the southeast of Brazil

The article describes the larva, pupa, adults and karyotype of Chironomus reissi, sp. n., which live on leaf detritus beds of small temporary waters in the southeast of Brazil. The species belongs to the pseudothummi cytocomplex with the chromosome arm combination AE, BF, CD, G.     

Subsidies for production of Chironomus calligraphus larvae (Chironomidae,Diptera) in the laboratory

The objective of this study was to evaluate the effect of temperature, type of mineral substrate, and type and amount of feed on the growth of Chironomus calligraphus as a subsidy for the production of Chironomus larvae to feed the fish. Egg production was evaluated at 22, 25, and 28?°C on fine (0.074–0.210?mm), or silty sand (0.002–0.250?mm), and no mineral substrate as control. For larval production, 0.10, 0.20, and 0.30?mg/larvae of fish feed, poultry feed, or turkey manure were used as food. The highest number of egg masses was produced when mineral substrates and 25?°C were used, but the higher number of eggs/mass was registered in the control. Therefore, the total number of eggs and hatched larvae produced was the same in the presence or in the absence of mineral substrates (p?>?0.05). The best results for survival and growth were recorded for larvae reared with fish or poultry food. The increase in the amount of feed was correlated with weight gain only for larvae fed with poultry feed, with no reduction in survival.     

A dated molecular phylogeny for the Chironomidae (Diptera)

We provide the first highly sampled phylogeny estimate for the dipteran family Chironomidae using molecular data from fragments of two ribosomal genes (18S and 28S), one nuclear protein‐coding gene (CAD), and one mitochondrial protein‐coding gene (COI), analysed using mixed‐model Bayesian and maximum likelihood inference methods. The most recently described subfamilies Chilenomyiinae and Usambaromyiinae proved elusive, and are unsampled. We confirm monophyly of all sampled subfamilies except Prodiamesinae, which contains Propsilocerus Kieffer, previously in Orthocladiinae. The semifamily Chironomoinae is confirmed only if Telmatogetoninae is included, which is closer to Brundin's original suggestion. Buchonomyiinae is excluded from Chironomoinae: it is a sister group to all remaining Chironomidae, conforming more to Murray and Ashe's argumentation. Semifamily Tanypodoinae is a grade and unsupported as monophyletic: the austral Aphroteniinae alone is sister to all Chironomidae (less Buchonomyiinae). Podonominae is weakly supported as the next sister group, in contrast to some estimates that place this subfamily as sister group to Tanypodinae alone. In Diamesinae, the southern African Harrisonini is confirmed as a member, but embedded within austral tribe Heptagiini, which is confirmed as sister to the undersampled Diamesini. Tribe Pentaneurini and ‘non‐Pentaneurini’ taxa are reciprocally monophyletic in Tanypodinae. Recent molecular findings concerning Podonominae are substantiated, with a monophyletic tribe Podonomini, Boreochlini forming a grade and Lasiodiamesa Kieffer placed as sister to all other Podonominae, but with uncertainty. In Orthocladiinae, a postulated two‐tribe system of Orthocladiini and Metriocnemini can be supported after exclusion of a Corynoneura group and a Brillia group, which is revealed as sister to Stictocladius Edwards. The marine Clunio Haliday and Thalassosmittia Strenzke & Remmert (given high rank in the past) are clearly embedded deep in Orthocladiinae. The finding of Shangomyia Sæther & Wang + Xyiaomyia Sæther & Wang as sister group to all other Chironominae justifies high rank, as their authors suggested. Pseudochironomini (untested by sampling shortfall) is sister to a monophyletic Tanytarsini (with a weakly supported inclusion of the enigmatic Nandeva Wiedenbrug, Reiss & Fittkau). The tribe Chironomini can be supported only by excluding Shangomyia + Xyiaomyia, and a postulated monophyletic clade comprising several taxa such as Microtendipes Kieffer, with six‐segmented larval antennae and alternate Lauterborn organs, that is sister group to Pseudochironomini + Tanytarsini. The tempo of diversification of the family, deduced by divergence time analysis (beast ), shows Permian origination with subfamily stem‐group origination from the mid–late Triassic to the early Cretaceous. Crown‐group origination ranged from Podonominae on a short stem originating in the mid Jurassic to long‐stemmed Aphroteninae from the late Cretaceous. Node dates allow inference of some vicariance via Gondwanan fragmentation, including certain nodes involving southern Africa.     

The role of glycogen during the ontogenesis of Chironomus anthracinus (Chironomidae, Diptera)

Large fluctuations in glycogen content were found in larvae, pupae and adults of Chironomus anthracinus (Zetterstedt) from the profundal zone of Lake Esrom, Denmark. In 2nd, 3rd and 4th instar larvae the glycogen concentration (expressed as percentage of dry weight) increased during periods of aerobic conditions to a maximum of 25%, but decreased in periods of hypoxia longer than two months to 10–12% in young larvae. A further decrease to about 5% took place, when moulting from 2nd to 3rd or from 3rd to 4th instar occurred after overturn. Prior to pupation the glycogen concentration was restored to 26–28%. The glycogen concentration approximated 22% in young pupae, but decreased during the pupal stage and newly hatched adults contained 12–15%. Finally, the glycogen store of both males and females was further reduced during the swarming period. Thus, glycogen seems to be an important energy source (1) during periods with hypoxic conditions, (2) during periods with high internal energy requirement such as ecdyses and metamorphosis, and (3) during the non-feeding adult life stage.     

Ecological synopsis for Chironomus flaviplumus (Diptera: Chironomidae) in the Jung-rang stream

In this study, we conducted field sampling to assess the relative influences of water and substrate quality on benthic macroinvertebrate communities living in the Jung‐rang stream, Korea. We collected macroinvertebrates and assessed physicochemical variables from three sites in the stream between May 2001 and January 2002. Sites were located approximately within 20 km from the headwater. The structure of the benthic macroinvertebrate communities may be strongly affected by the physical conditions inherent to the environment in which they live. In this stream, we detected profound differences in water temperature (18 ～ 19.75°C), the concentrations of suspended solids (3.935 ～ 7.87 mg/L), and demand for chemical oxygen (10.575 ～ 14.425 mg/L). Nonylphenol concentrations ranging from 0.375 to 0.55 ng/mL were found in the water, and the sediments were measured to contain between 2.45 and 3.425 ng/mL. We identified a total of 20 macroinvertebrate species, including seven species of Chironomidae, the most abundant of which was Chironomus flaviplumus. At none of the sites did we find any significant differences in the structure of the communities. Using canonical correspondence analysis for the relation of species and environmental variables, chemical oxygen demand and suspended sediment gradients (SS) had significant preferences for site 1 with SS. The results of our study suggest that physico‐chemical variables exerted complex effects on the structure of the benthic community in the Jung‐rang stream. This study supports the contention that physico‐chemical analyses as well as community analysis are valuable tools to assess the effect of pollution on the ecological condition of a stream. Chironomids, in particular, showed a high degree of tolerance against contaminants.     

Chromosomal variability in natural populations of Chironomus cingulatus meigen (Diptera, Chironomidae)

Chromosomal polymorphism has been studied in seven natural populations of Chironomus cingulatus from Western Europe, Western Siberia, and the Republic of Sakha (Yakutia). The banding sequences pool of the species includes 15 banding sequences. Chromosomal polymorphism was revealed in five out of seven chromosomal arms. Arm B is the most polymorphic with four banding sequences. There are three banding sequences in arm A. Arms D, E, and G have two banding sequences. None of the chromosome rearrangements were revealed in arms C and F. The populations of C. cingulatus differ clearly in their number and frequency of banding sequences, which indicates that different gene sequences are adaptive in different populations.     

Biological and molecular responses of Chironomus riparius (Diptera,Chironomidae) to herbicide 2,4-D (2,4-dichlorophenoxyacetic acid)

2,4-Dichlorophenoxyacetic acid (2,4-D) is an agricultural contaminant found in rural ground water. It remains to be determined whether neither 2,4-D poses environmental risks, nor is the mechanism of toxicity known at the molecular level. To evaluate the potential ecological risk of 2,4-D, we assessed the biological parameters including the survival rate, adult sex ratio of emerged adults, and mouthpart deformities in Chironomus riparius after long-term exposure to 2,4-D. The larvae were treated with 0.1, 1 or, 10 μg L^? 1 of 2,4-D for short- and long-term exposure periods. The sex ratio was changed in C. riparius exposed to only 10 μg L^? 1 of 2,4-D, whereas mouthpart deformities were observed as significantly higher in C. riparius exposed to 0.1 μg L^? 1 of 2,4-D. Survival rates were not significantly affected by 2,4-D. Furthermore, we evaluated the molecular and biochemical responses of biomarker genes such as gene expression of heat shock proteins (HSPs), ferritins and glutathione S-transferases (GSTs) in C. riparius exposed to 2,4-D for 24 h. The expressions of HSP70, HSP40, HSP90 and GST levels in C. riparius were significantly increased after exposure to a 10 μg L^? 1 concentration of 2,4-D, whereas ferritin heavy and light chain gene expressions were significantly increased at all concentrations of 2,4-D exposure. Finally, these results may provide an important contribution to our understanding of the toxicology of 2,4-D herbicide in C. riparius. Moreover, the 2,4-D-mediated gene expressions may be generated by 2,4-D is the causative effects on most probable cause of the observed alterations. These biological, molecular and morphological parameters and the measured parameters can be used to monitor 2,4-D toxicity in an aquatic environment.     

Locomotory modes in the larva and pupa of Chironomus plumosus (Diptera, Chironomidae)

The locomotory kinematics of Chironomus plumosus larvae and pupae were investigated in order to determine how different locomotory techniques may be related to (a) possible underlying patterns of muscle activation and (b) the particular lifestyles and behaviours of these juvenile stages. Larvae display three independent modes of motile activity: swimming, crawling and whole-body respiratory undulation. Swimming and respiratory undulation involve the use of metachronal waves of body bending which travel in a head-to-tail direction. Whereas swimming is produced by side-to-side flexures of the whole body, respiratory undulation employs a sinusoidal wave. Crawling appears to result from an independent programme of muscle activation. Instead of a longitudinally transmitting metachronal wave of body flexure, a simultaneous arching of the body, combined with the alternating use of the abdominal and prothoracic pseudopods as anchorage points, produces a form of locomotion analogous to caterpillar-looping. Larval swimming has a set speed and rhythm and is an 'all-or-nothing' locomotory manoeuvre, but the neural programme controlling larval crawling is adaptable; switching from a less to a more slippery substrate resulted in a shorter, faster stepping pattern. The pupa displays two swimming modes, somersaulting and eel-like whole-body undulation, the former being principally a brief, escape manoeuvre, the latter being a faster form of locomotion employed to deliver the pupa to the surface prior to adult emergence. Comparison with the pupa of the culicid Culex pipiens shows that this insect also uses the somersault mechanism but at a higher cycle frequency which produces a faster swimming speed. This appears to be related to differences in lifestyle; the surface-living culicid pupa is exposed to greater predator threat than the bottom-dwelling chironomid pupa, and consequently needs a faster escape.     

Cytogenetic studies of Chironomus circumdatus from India (Diptera: Chironomidae)

Chironomus circumdatus is one of the most common and wide-spread species in India. Cytogenetic studies pertaining to the mitotic and polytene chromosomes, nucleolar organizer regions, C-banding and naturally occurring chromosomal polymorphism have been carried out for the first time in this Indian species. Altogether seven inversions comprising six paracentric and one pericentric are detected in the Indian populations. The distribution of inversions in relation to different environmental conditions is discussed.