ABSTRACT
Chironomids have been widely used in environmental monitoring and toxicity assays. Assessment criteria for chironomids range from the molecular-biochemical level to the population level. However, the use of markers to evaluate cellular and anatomical changes in organs and systems of individuals under contaminant exposure is still incipient. In this study, we conducted a histological description of the main systems and organs of immature Chironomus columbiensis. We used fourth instar larvae, obtained from a standardized culture, and confirmed this state through morphological and molecular methods. Larvae were fixed in Duboscq solution for insects during 48 hr, dehydrated in increasing dilutions of ethanol, and embedded and mounted in historesin to obtain 3 µm sections. The digestive, nervous, excretory, and integumentary systems and the fat body and testicles were analyzed through longitudinal and transversal sections, stained with hematoxylin-eosin (HE), and further photographed and described through light microscopy. This is the first description of internal morphology performed for C. columbiensis and can help in future histopathological analysis, because through bioaccumulation some of these organs are contaminant targets. It can even be a great taxonomic tool, since the characteristics of the histological pattern of C. columbiensis presented differences compared to other descriptions made in chironomids.
Subject(s)
Chironomidae/anatomy & histology , Environmental Monitoring/methods , Larva/anatomy & histology , Animals , Ecosystem , Histological Techniques , Specimen Handling , Staining and LabelingABSTRACT
ABSTRACT Changes in physiology of the nervous system and metabolism can be detected through the activity of acetylcholinesterase (AChE), alpha esterase (EST-a) and beta esterase (EST-ß) in chironomids exposed to pollutants. However, to understand the real effect of xenobiotics on organisms, it is important to investigate how certain factors can interfere with enzyme activity. We investigated the effects of different temperatures, food stress and two steps of the enzymatic protocol on the activity of AChE, EST-a and EST-ß in Chironomus sancticaroli. In experiment of thermal stress individuals from the egg stage to the fourth larval instar were exposed to different temperatures (20, 25 and 30 °C). In food stress experiment, larvae were reared until IV instar in a standard setting (25 °C and 0.9 g weekly ration), but from fourth instar on they were divided into groups and offered different feeding regimes (24, 48 and 72 h with/without food). In sample freezing experiment, a group of samples was processed immediately after homogenization and another after freezing for 30 days. To test the effect of centrifugation on samples, enzyme activity was quantified from centrifuged and non-centrifuged samples. The activity of each enzyme reached an optimum at a different temperature. The absence of food triggered different changes in enzyme activity depending on the period of starvation. Freezing and centrifugation of the samples significantly reduced the activity of three enzymes. Based on these results we conclude that the four factors studied had an influence on AChE, EST-a and EST-ß and this influence should be considered in ecotoxicological approaches.
ABSTRACT
Low malathion concentrations influence metabolism in Chironomus sancticaroli (Diptera, Chironomidae) in acute and chronic toxicity tests. Organophosphate compounds are used in agro-systems, and in programs to control pathogen vectors. Because they are continuously applied, organophosphates often reach water sources and may have an impact on aquatic life. The effects of acute and chronic exposure to the organophosphate insecticide malathion on the midge Chironomus sancticaroli are evaluated. To that end, three biochemical biomarkers, acetylcholinesterase (AChE), alpha (EST-α) and beta (EST-β) esterase were used. Acute bioassays with five concentrations of malathion, and chronic bioassays with two concentrations of malathion were carried out. In the acute exposure test, AChE, EST-α and EST-β activities declined by 66, 40 and 37%, respectively, at 0.251 µg L-1 and more than 80% at 1.37, 1.96 and 2.51 µg L-1. In chronic exposure tests, AChE and EST-α activities declined by 28 and 15% at 0.251 µg L-1. Results of the present study show that low concentrations of malathion can influence larval metabolism, indicating high toxicity for Chironomus sancticaroli and environmental risk associated with the use of organophosphates.
