Design of molecular markers for identification of species of the genus Chironomus (Diptera,Chironomidae)

Accurate identification and differentiation of species of the genus Chironomus based on their morphological features is a difficult problem. Unambiguous species identification by means of molecular markers is possible at any stage of the life cycle. Polymerase chain reaction (PCR) with species-specific primers was used to develop molecular markers (amplicons) for identification of Chironomus piger, Ch. dorsalis, and Ch. pseudothummi. Nucleotide sequences of the internal transcribed spacer region (ITS) of the locus coding for ribosomal RNA were used to design species-specific primers for these target species. Each of the species-specific primer pairs yielded species-specific amplicons (molecular markers) only with the DNA of target species: Ch. piger, Ch. dorsalis, and Ch. pseudothummi. Test PCRs with the DNA of eighteen Chironomus species confirmed the specificity of the primers obtained. The molecular markers produced in PCR with the designed species-specific primers permit reliable identification of Ch. piger, Ch. dorsalis, and Ch. pseudothummi and their differentiation from other species of the genus Chironomus.     

A review of the genusChironomus (Diptera,Chironomidae)

Two cytologically distinguishable species have been found in material ofChironomus australis. These species have been calledCh. australis andCh. duplex. Both belong to thepseudothummi-cytological grouping because they have the chromosome arm combinations AE, BF, CD, G.Ch. duplex shows a modified arm pattern due to a tandem fusion of arm G to arm E.—The banding patterns of the polytene chromosomes of the two species are compared to each other and to the Australian standard species,Ch. oppositus. Ch. australis is very close cytologically toCh. oppositus, whileCh. duplex, which is considered a derived species because of the tandem fusion, shows a number of inversion differences from the morphologically similarCh. australis. Ch. duplex is polymorphic for six inversions, four of which are simple inversions, the other two are complex involving also the transposition of some bands.Ch. australis appears to be monomorphic.     

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 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.     

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.     

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.     

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.     

Succession of Chironomus Meigen Species (Diptera,Chironomidae) in Newly Filled Ponds

The colonization by Chironomus spp. of two ponds filled after a dry period was investigated. Field data were collected by means of quantitative benthic samples and emergence traps. C. melanescens. C. annularius, C. riparius, C. piger, C. luridus, C. obtusidens, C. plumosus, C. dorsalis and Camptochironomus tentans were determined cytotaxonomically in ponds investigated. C. melanescens and C. annularius appear to be the most characteristic species colonizing in high numbers in newly filled ponds. Larvae of C. annularius grew more slowly in ponds compared with the former one which resulted in a characteristic succession of both larval types. Most species mentioned disappeared completely from the ponds investigated after the metamorphosis of a single cohort. They were replaced by C. plumosus which was often attended by C. annularius in smaller ponds.     

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.     

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.     

Karyotypic features of Chironomus entis and Chironomus borokensis (Diptera, Chironomidae) from Lake Kotokel (Lake Baikal Basin)

Karyotypes and chromosomal polymorphism of sympatric sibling species Chironomus entis and Chironomus borokensis from Lake Kotokel were studied. Five inversion banding sequences were found in C. entis, including p’entA16, p’entB9, and p’entF6 that are new for the species; in C. borokensis, four inversion banding sequences were revealed, borA4 being new for the species. The level of chromosome polymorphism in C. entis and C. borokensis can be considered similar: 54.2 and 53.8% heterozygosity, 0.67 and 0.73 inversions per individual, respectively. The new data are compared with those on C. entis population from Lake Kotokel obtained in 1981 and those of C. borokensis from Lake Dukhovoe (Lake Baikal basin) obtained in 1983.     

Karyotype of the midge Chironomus heterodentatus Konstantinov from the group Obtusidens (Diptera, Chironomidae)

The karyotype of Chironomus heterodentatus, belonging to the obtusidens-group described by Konstantinov from the Volga in 1956, was studied in detail. Combinations of chromosomal arms are AB, CD, EF and G (cytocomplex thummi). The Ch. heterodentatus karyotype cleary differs from those of other members of the obtusidens-group with a species-specific banding pattern in arms A, B, C and D. Chromosomal polymorphism on homo- and heterozygous inversions was found in arms, A, B, D, E and G. 19 inversion banding sequences and their 27 genotypic combinations have been recorded. The shortest arm G is highly polymorphic. Heterozygotes on the Balbiani ring activity were found in arm G along with homo- and heterozygotes on inversions. B-chromosomes with a frequency equal to 2.7-25.0% were recorded in some Volga populations.     

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.     

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.     

Function of Ventral and Lateral Processes in Larvae of Chironomus (Diptera, Chironomidae)

Oxygen consumption by larvae of five Chironomus species from the Plumosus group belonging to different larval forms was studied. Larvae of C. agilis ( f. l. reductus) were unable to utilize oxygen at its concentrations lower than 1 mg/l. Larvae of C. plumosus ( f. l. plumosus) used oxygen from water at concentrations higher than 0.7 mg/l; pupas of this species and larvae of C. borokensis ( f. l. plumosus) utilized oxygen at concentrations lower than 0.4 mg/l. C. entis and C. muratensis (both-- f. l. semireductus) utilized oxygen from water at its concentrations up to 0.05 mg/l. The lack of correlation between the rate and ability of larvae to use oxygen at low concentrations and the degree of development of processes allows suggesting that these structures are not additional respiratory organs, as is commonly accepted. At the same time, ability of species to inhabit under conditions of oxygen deficiency correlates directly with the size of the processes. Therefore, ventral and lateral processes are suggested to perform a function of excretion of anaerobic metabolism metabolites.     

Karyotype and morphology of larvae of Chironomus (Chironomidae, Diptera) from the Caspian Sea

Karyotype and morphology of Chironomus sp. larvae from the Caspian Sea have been described. 2n = 8. Chromosomal arm combination is AB, CD, EF, G (cytocomplex thummi). All chromosomes display conjugation of homologues. Centromere areas are of s-type. The nucleolar organizer and two Balbiani rings are disposed in arm C, and another Balbiani ring is in arm B. Half larvae have heterozygotic paracentric inversions in arm D. The larva belongs to salinarius form. One scleritis is colorless, an occipital scleritis is light and irregularly colored. Premandible has 4-5 cogs. Epipharingeal ridge has 18-23 dens. This new species is close to salinarius group, especially to Ch. albidus Konst., but differs from it at the larva stage in the structure of non-massive basal antenna segment, in the presence of large quantity of premandible denses, in light color of occipital scleritis, and in a series of morphometric rates. A similarity berween Ch. paraalbidus and Ch. albidus, on the one hand, and Boeotendipes, on the other one, has been found out.