Chapter 1 Condition Index, Behavioural Study and Oxygen Consumption
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Chapter 1 Condition Index, Behavioural Study and Oxygen Consumption Chapter 1: Condition Index, Behavioural Study and Oxygen Consumption Chapter 1: Condition Index, Behavioural Study and Oxygen Consumption 1.1 General Introduction Class Bivalvia of phylum Mollusca includes soft-bodied forms which are enclosed within a bivalve shell. Molluscs form the second largest group of animals after insects in the whole animal kingdom. They are highly adaptive and occupy all possible habitats except aerial. Most family members of this class are mussels, clams, oysters etc. Lamellidens are the most common freshwater mussels found in India, inhabiting the surfaces layers of the muddy beds of the lakes, rivers and streams. They form one of the largest groups in terms of biomass amongst the filter feeding organisms in many freshwater ecosystems (Mickael, 1995). They are important calcium and carbon accumulators; they link primary producers (bacteria and phytoplankton) with higher organisms in aquatic food-chains. The freshwater mussel plays a vital role in freshwater environment. They are often described as sessile, primary consumers which can be used to assess different routes of contaminant exposure (Farris, 2007). The freshwater mussels, Unionid mussels in particular, are more sensitive to chemical exposure and a variety of other environmental stressors, as compared to other organismal groups (Surwase, 2009). They need to be considered as a part of whole freshwater community. Mussels are filter-feeding sedentary species, especially prone to the accumulation and concentration of contaminants. As filter feeders, their vulnerability to different contaminants, chemicals and other pollutants in aquatic habitats seem to be high. Freshwater bivalves are highly sensitive to toxic contaminants. Exposure assessment is essential to understanding the potential effects of contaminants to non-target animal populations. Recent surveys have indicated declining diversity and density of mussels because of different contaminants in aquatic ecosystems. Mussels readily bio accumulate both organic (Moore M, 1985) and metal pollutants (Vairengo et.al., 1985) thereby rendering them more accurate in situ bio-indicators of pollution than fish (Cajaraville et. al., 2000). 12 Chapter 1: Condition Index, Behavioural Study and Oxygen Consumption Human beings also consume mussels and therefore, any pollutant which can accumulate in mussel tissues has the potential to enter higher tropic level organisms via the food chain. Mussels are sessile, which make them useful as bio-indicators since they stay in one place and their health status yields information relevant to a particular location (Dondero et.al., 2006). Mussels’ lifestyle is stationary, filter feeding, inhabiting the benthic environment where pollutants usually end up and accumulate. Therefore, a mussel provides many advantages as a model to assess toxic effect of environmental pollutants in organisms (Al Amri et. al., 2012). The impact of aquatic pollutants is thought to be significantly different in various organs/tissues along with their own biological function variability. For example, gill tissue and digestive gland of mussels play important roles in food collection, absorption and digestion. Chronic exposure of mussels to pollutants in water and sediments may ultimately impair their nutrient absorption ability, compromise their growth and reproduction (Smital et. al., 2004). Owing to high filtering capacity of mussels, gill tissues are not only continuously in contact with pollutants in waters, but they may also concentrate pollutants contained therein due to the remarkably high volume of water that they filter (Au, 2004). Pollution is an alarming problem that the mankind is facing today. We unknowingly do a lot of activities that terribly ruin the nature. Water contamination is one of the severe factors that affect the aquatic ecosystem. Aquatic environment is continuously being contaminated with metals and toxic chemicals from industrial, agricultural and domestic activities. Heavy metal contamination interferes with ecological balances of an ecosystem and produces devastating effects on environment quality (Farombi et. al., 2007). The freshwater mussels play a vital role in food chain and today, sadly, are among the most threatened aquatic species in the world. One of the major issues implicated in this decline is water pollution. Fresh water mussel populations have suffered a lot because of habitat disturbances & commercial demands. They need to be considered as a part of whole freshwater community, particularly because they link up various aquatic food chains involving the edible fish, which form an important resource of protein for the human population. Since mussels exist in direct contact with contaminated aquatic 13 Chapter 1: Condition Index, Behavioural Study and Oxygen Consumption sediments and are exposed to water-borne contaminants, they represent interesting specimens in eco-toxicological studies. Therefore, mussels have been used extensively as sensitive bio-indicators for aquatic pollutants (Livingstone et.al., 1992) they represent interesting specimens in eco-toxicological studies since they exist in direct contact with contaminated aquatic sediments and they are exposed to water-borne contaminants. 1.2 Aim and objectives of the present study There are several advantages of mussels, as they occur in most of the water bodies and they have stationary, filter feeding life style, inhabiting benthic environment, where different contaminants accumulate. Mussels readily bio accumulate both organic (Moore, 1985) and metal (Viarengo, 1985) pollutants. The present work is proposed to study biological effects of contaminants like cupric chloride dihydrate, PAH (Anthracene) individual and combined exposure of both together for seven (T1) and fourteen days (T2) on mussels. The objectives of the present study: 1. Estimation of level of metals in water & in tissues of bivalves, collected from different reservoirs of water 2. Assessment of health of animals at the time of collection from their natural reservoirs 3. Analyses of acute toxicity of contaminants such as - Cupric chloride di-hydrate (A), Anthracene (B) and combined effect of mixture (C=A+B) on Lamellidens corrianus in laboratory 4. Periodic observations of behavioural changes on exposure to A, B and C 5. Determination of rate of oxygen consumption 6. Determination of neurotoxicity by analysis of AChE activity 7. Determination of alterations in biomarkers of oxidative stress, such as MDA, SOD, CAT, GST and GR 8. Assessment of histopathological changes in gills after exposure to different contaminants 9. Estimation of Metallothionein content upon exposure to cupric chloride di-hydrate (Cucl2.2H20) in bivalves 14 Chapter 1: Condition Index, Behavioural Study and Oxygen Consumption 10. Quantification of metal bioaccumulation in soft tissues of bivalve 11. Assessment of genotoxicity in the gill cells by using comet assay 12. Detection of chromosomal damage with the help of micronuclei formation, MN test 13. Assessment of recovery potential of animals following the A, B and C exposures 1.3 Selection of experimental animal The present study deals with the investigation and assessment of toxicity of cupric chloride dihydrate and anthracene individual and their combined effects of both together on non-target fresh water bivalve Lamellidens corrianus in laboratory, this species selected for the present study because it fulfils most of the criteria for a standard test species (Fishes, mollusks, and crustaceans) as described by Adelman and Smith (1976). i. Test species should be capable of being maintained in the laboratory, with healthy conditions for at least one month, ii. It must be available throughout the year with proper and required size, iii. It should be easy in handling, collection and transportation, iv. It should be available in sufficient numbers for repeating the toxicity tests, v. It must have a constant response to toxicants tested under similar controlled conditions of exposure. It has been observed that bivalves are the most suitable and are easily harvestable from the water bodies, they are widely used as ecological indicators because they are sensitive to ecological stress and being filter feeders they accumulate contaminants from water bodies which makes them highly suitable as test organism. Since L. corrianus fulfils above criteria and in environmental conditions it gets constantly exposed to toxic contaminants directly or indirectly through food chain, hence it is considered as an ideal test species in toxicological studies. 15 Chapter 1: Condition Index, Behavioural Study and Oxygen Consumption The taxonomic classification of L. corrianus is as under, 1.3.1 Systematic Position Phylum: Mollusca Class: Bivalvia Subclass: Paleoheterodonta Order: Trigoinoida Superfamily: Unionoidea Family: Unionidae Subfamily: Ambleminae Tribe: Amblemini Genus: Lamellidens Species: corrianus Figure 1.1: Lamellidens corrianus 16 Chapter 1: Condition Index, Behavioural Study and Oxygen Consumption 1.3.2 Collection and Maintenance in the laboratory The freshwater mussels L. corrianus, were collected from the reservoirs around Pune and Ozar Tal. Junner Dist. Pune (shell- length 7-9 cm). This species is found at the bottom of ponds, rivers, lakes and other fresh water bodies. They are mostly found buried in mud in stagnant and slow running water hence can be easily collected. The water from the site of collection was examined to ascertain any traces