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Technique for mass rearing of Indian Chironomus species

Authors:
  • Savitribai Phule Pune University Pune India
NATH& GODBOLE:187-193 Studia dipterologica 5 (1998) Heft 2
Technique for mass rearing of Indian Chironomus species
(Diptera, Nematocera, Chironomidae)
[Eine Technik zur Massenzucht von indischen Chironomus-Arten
(Diptera, Nematocera, Chironomidae)]
by
Bimalendu B. NATH and Narayan N. GODBOLE
Pune (India)
Key words
At present, no standard methodologies are available to colonize and rear tropical
midges of the family Chironomidae (Diptera). We present here a simple. inexpensive
laboratory method of mass rearing Indian Chirono/lllls, based on locally available
food. Morphometric analysis to monitor larval growth has been carried out to establish
suitability of the rearing methodology. The rearing technique described in this paper,
provides a continuously breeding population of three Indian species of Chirono/lllls.
mass rearing, Chironomidae, Chirono/lllls, India
Abstract
Stichwiirter
Derzeit existiert keine Standartmethode, mit der tropische Zuckmlicken (Diptera,
Chironomidae) unter Laborbedingungen zu halten und zu vermehren sind. Es wird
eine einfache, wenig kostenintensive Labormethode zur Zucht yon Chirono/lllls-Ar-
ten der indischen Fauna vorgestellt, die auf derVerwendung eines einheimischen Zucht-
substrates beruht. Eine Uberwachung des Wachstums der Larven li\;)erMessungen der
Kopfkapselbreite wurde angewendet, urn die Brauchbarkeit der Zuchtmethode abzu-
sichern. Die hier geschilderte Methode wurde erfolgreich bei der kontinuierlichen
Haltung yon drei indischen Chirollollllls-Arten verwendet.
Massenzucht, Chironomidae, ChiroI:.0/llIIS,(ndien
Zusammenfassung
Introduction
Chironomid midges constitute a major biotic component in most of the freshwater eco-
system (OLIVER1971, PINDER1986). For decades, a great deal of research had been done
in cytogenetics and pollution biology using Chironomus as a system (MURRAY1980,
ARMITAGEet al. 1994). In the past, several workers described methods of rearing that
suited to their experimental conditions. It was however experienced that none of the
available methods were suitable either because of nonavailability of food components or
because of different conditions required by tropical midges. Hence a new method has
been evolved suitable for maintaining tropical Indian midges as a continuous culture
under laboratory con~itions.
A method for the propagation of Chironomus tentans FABRICIUS,1805, was reported by
SADLER(1935). However, alternative methods were utilized by subsequent workers for
rearing C. tentans (BENTIVEGNA& COOPER1993, FIRLING& KOBILKA1979, MEYERet al.
1983). BIEVER(1965) studied different species of Chironomus, namely C. attenuatus
WALKER, 1848; C. fulvipilus REMPEL,1939; C. monochromus VANDERWULP, 1874; C.
stigmaterus SAY, 1823; C. califomicus JOHANNSEN,1905 and described techniques for
rearing in the laboratory (reviewed by SINGH& MOORE1985). BIEVER(1965) also de-
scribed composition of food, conditions for oviposition and designing of rearing cages.
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... Chironomus ramosus, a common Indian tropical midge, inhabits freshwater ecosystems and is adapted to thrive under adverse environmental conditions (Armitage et al., 1995;Nath and Godbole, 1998). These primitive groups of dipteran insects, show tolerance to wide range of biotic (hypoxia, heat) and abiotic (water pollutants) stress factors (Choi et al., 2000;Yoshimi et al., 2002;Callaghan et al., 2002;Haas et al., 2005;Hassell et al., 2006;Nowak et al., 2007;Vogt et al., 2007). ...
... Thirty days old fourth instar larvae of the tropical insect species of midge C. ramosus were reared in insectary of Bhabha Atomic Research Centre, Mumbai, India. The laboratory cultures were maintained using mass rearing technique (Nath and Godbole, 1998) at 25 ± 2°C with relative humidity 75 ± 10% and 14 h:10 h light and dark photoperiod cycle. ...
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A battery of enzymes from the eukaryotic antioxidant defense system was measured in salivary gland and in whole body extract of fourth instar larvae of Chironomus ramosus with an objective of finding any clue for the dipteran insect's capacity to tolerate heavy doses of ionizing radiation. Levels of activity of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GSH-Px) were quantified in 30 days old larvae exposed to LD 20 dose of gamma radiation. Compared to controls, activity of Cu,Zn-SOD increased 3 to 4 fold and catalase 2 fold in response to ionizing radiation stress, while activities of GR and GSH-Px enzymes were decreased. Among the other SOD isoenzymes, our results showed comparable levels of Mn-SOD and Cu,Zn-SOD activity in control and irradiated groups of larvae. The increase in levels of the Cu,Zn-SOD isoenzyme was also confirmed by Western blot and zymography supported by densitometric quantification. No evidence of Fe-SOD was found in C. ramosus larvae. These findings could help to explain the persistence of natural populations of Chiro-nomus in radioactively contaminated regions.
... Previously, several works on HSPs and heat shock inducible chromosomal loci were reported mainly in two European species, namely, Chironomus tentans Fabricius and Chironomus thummi (Kieffer) (Tanguay and Vincent 1981;Lezzi 1984;Carretero et al. 1991;Sass 1995;Martinez et al. 1997;Morcillo et al. 1997) and in one Indian species, Chironomus striatipennis Kieffer (Nath and Lakhotia 1989). In the present study, a tropical Indian species, Chironomus ramosus Chaudhuri, Das and Sublette, colonized in the laboratory (Nath and Godbole 1998) has been chosen to study behavioural response to heat shock. ...
... Larvae were originally obtained from an isofemale line of S/S wild type population following a breeding scheme described by Hardikar and Nath (2001). All developmental stages were maintained in the laboratory by using a mass rearing technique (Nath and Godbole 1998) and fourth instar larvae from the synchronous culture were used for the present study. ...
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All living cells respond to temperature stress through coordinated cellular, biochemical and molecular events known as “heat shock response” and its genetic basis has been found to be evolutionarily conserved. Despite marked advances in stress research, this ubiquitous heat shock response has never been analysed quantitatively at the whole organismal level using behavioural correlates. We have investigated behavioural response to heat shock in a tropical midge Chironomus ramosus Chaudhuri, Das and Sublette. The filter-feeding aquatic Chironomus larvae exhibit characteristic undulatory movement. This innate pattern of movement was taken as a behavioural parameter in the present study. We have developed a novel computer-aided image analysis tool “Chiro” for the quantification of behavioural responses to heat shock. Behavioural responses were quantified by recording the number of undulations performed by each larva per unit time at a given ambient temperature. Quantitative analysis of undulation frequency was carried out and this innate behavioural pattern was found to be modulated as a function of ambient temperature. Midge larvae are known to be bioindicators of aquatic environments. Therefore, the “Chiro” technique can be tested using other potential biomonitoring organisms obtained from natural aquatic habitats using undulatory motion as a behavioural parameter.
... Larvae collected from the Mula river were validated for taxonomic identification using morphological and cytotaxonomic keys [26,27] and reared in non-toxic plastic tubs (Ø = 35 cm) containing sterilized beach sand at the bottom (S2A Fig). The rearing procedure was described previously [28,29]. In brief, to initiate rearing,~100 late fourth instar larvae were transferred to a separate plastic tub. ...
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... Chironomus ramosus larva (acetocarmine staining) showing Balbiani ring regions identified using a previously published cytological reference map (Nath and Godbole, 1998). Fig. 4. Venn diagram constructed to illustrate the diversity and occurrence of silk proteins among the three well studied Chironomus species (C. ...
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The larvae of the aquatic Chironomid midge (Diptera, Chironomidae) are known to survive in freshwater habitats exposed to stress especially that of hypoxic stress. Hence, a study of the effect of alterations in the levels of dissolved oxygen on the physiology of the larvae could help in establishing them as potential sensitive bio indicators for monitoring such freshwater habitats. A simple inexpensive and non-cumbersome experimental setup unlike commercially available sophisticated set-ups was designed and fabricated in the lab. The validation of this set-up was carried out and alterations in the hemoglobin levels in the larvae of the tropical midge species Chironomus ramosus exposed to hypoxic conditions were studied. It was observed that there was an increase in the level of hemoglobin on exposure to hypoxic conditions and the findings were further validated by studying the expression of hemoglobin gene at various time points during exposure to hypoxia. These findings suggest that the device designed could be used as an inexpensive and effective method for generating and maintaining the desired levels of dissolved oxygen as compared to several of the chemical and physical methods generally used. Further, this would be useful in studies to be done for the monitoring of hypoxia in freshwater habitats using Chironomus hemoglobin as a biomarker.
... Rearing of midge species in the insectary is essential for scientific investigations. Above all, midge species was maintained in the insectary following Nath and Godbole (1998), Habashy (2005), Nath et al. (2009) and Bhaduri et al. (2012). Midge larvae procured from the insectary were used as vehicle for delivering probiotics during experimental period. ...
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... 73 ° 51 ′ 48.06 ′ ′ E) inhabited by the natural population of C. ramosus . Culture was maintained by mass rearing technique (Nath and Godbole 1998) at 25 Ϯ 2 ° C temperature, relative humidity of 50 Ϯ 10%, and a photoperiod of 14 h light and 10 h darkness in the insectary of Bhabha Atomic Research Centre (BARC), Mumbai. Th irty-day-old fourth instar larvae were put in a glass beaker containing tap water and exposed to a dose of 2200 Gy a lethal dose for 20% of the midge population (LD 20 ) determined by dosimetric studies for the determination of percent mortality of irradiated larvae followed by Probit analysis (Datkhile et al. 2009a) from a cobalt 60 Co source (dose rate 5.5 Gy/min, Gamma Cell 220, Atomic Energy of Canada Ltd, Ottawa, Canada). ...
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