Toxicity and Histopathological effect of different organic waste on the earthworm (Eudrillus eugeniae and Eisenia fetida) under laboratory conditions

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ISSN No.2347-2901(Print) 2347-291X(Online) International Journal of Ethnomedicine and Pharmacological Research journal homepage: www.ijepr.

Eisenia fetida) under laboratory conditions Vimala Bangarusamy 1*,S.Karpagam 1 and P. Martin 2 1 Department of Botany, Queen Mary s College (autonomous), Chennai-600 004.

1 ISSN No (Print) X(Online) International Journal of Ethnomedicine and Pharmacological Research journal homepage: by the International Journal of Ethnomedicine and Pharmacological Research Toxicity and Histopathological effect of different organic waste on the earthworm (Eudrillus eugeniae and Eisenia fetida) under laboratory conditions Vimala Bangarusamy 1*,S.Karpagam 1 and P. Martin 2 1 Department of Botany, Queen Mary s College (autonomous), Chennai Department of Zoology, Govt. Arts College for Men, Nandanam, Chennai History of Article: Received 11March 2014 Revised from 18 March 2014 Accepted 21 March 2014 Available online 25April 2014 INTRODUCTION Soil is a pivotal component of the environment and it support diverse and numerous life form among which earthworm are considered to be representative organisms. Soil is also a medium for the spread of pollutants and plays a critical role in determining the toxicity of pollutants. Soil dwellers such as earthworm are inevitably subject to the toxicity effect of pollutants. On the other hand earthworms also have an impact on soil properties, structure and pollution. All these form the complex links between soil, earthworm and environmental pollution (Yasmin et al., 2007). Earthworms called the Intestine of earth by Aristotle are very important soil organisms that aid in the ABSTRACT The toxicity and histopathological effect of the industrial waste to the earthworm species Eudrillus eugeniae and Eisenia fetida were studied under laboratory conditions. Earthworm species were exposed to different organic waste (Garbage, Sugarcane baggase, Paper and polythene, Textile dye sludge, Herbicide) for 40 days and the mortality was recorded every 10 days. In addition one sections of the worm were made after 96 hours for microscopic examination. There was no mortality in the control group but in the experiment groups throughout the study. The histopathological manifestations that textile dye sludge and herbicide waste gradually increase damages of tissue. The soft peritoneal membrane lining intestinal epithelium is more affected. The result of the study showed that both mortality and histopathology data could be used in environmental risk assessment of textile dye sludge and herbicide. Keywords: Eudrillus eugeniae, Eisenia fetida and organic waste Page 18 decomposition of all organic matter. It represents a major component of the soil biomass (Townsend Lee et al., 2007). They are also important contributors to the recycling of carbon and nitrogen in the ecosystem. They make them one of the most suitable bio indicators organisms for testing chemicals in soil (Kavitha et al., 2008). All human activities generate some kind of byproducts of wastes which are apparently of no use to us and have to be discarded (Reinecke et al., 2007). Rapid urbanization, industrialization, unplanned population growth,misuse and abuse of the environment have led to an increased accumulation of solid wastes materials (Padma et al., 2002).The solid wastes produced by the town and cities are mainly the organic wastes causes fly breading,rodent menace, transmission of pathogens,soil * Corresponding author: Department of Botany, Queen Mary s College (autonomous), chennai , Tamilnadu, India vimalaphd@gmail.com, s.karpagam98@gmail.com.

2 pollution,bad odors, surface and ground water pollution (Joshi et al., 2008). Lack of awareness among the people, lack of cooperation from the public and lack of funds are the main causes for the improper disposal of solid wastes in towns and cities (Yasmin et al., 2010). The vermicomposting of the organic waste turns them into valuable agricultural inputs and minimize environmental problems (Rao, 2004). The term vermicomposting is coined from the Latin word vermis meaning to be worms. Vermicomposting a mesophilic process called by earthworms, Involves ingestion, digestion and absorption of organic waste followed by excretion of castings through the worms metabolic systems, enhancing the levels of plant nutrients of organic waste during the biological activities (Edward et al., 1988). The most frequently used earthworm species for vermicomposting are Eudrillus eugeniae and Eisenia fetida. The earthworm Eudrillus eugeniae is commonly known as African worm or Night crawls. It is a large composting worm, less tolerant to cold temperature and best suited to tropical conditions. Eisenia fetidia is commonly found in compost heaps, forests, gardens, under stones and logs. Eisenia fetida an epigeic earthworm species, feeds on inorganic wastes (Garg et al., 2005). Hence this study was undertaken with view to investigating the toxicity of different organic waste to two earthworm species, Eudrillus eugeniae and Eisenia fetida as well as histopathological effects. MATERIAL AND METHODS The soil collected from agriculture land in padappai, Chennai, Tamil Nadu. The soil was air-dried, ground and screened through a nylon fiber sieve to remove stones, plant root and other large particles. Two type of earthworm species were collected from Hand in Hand organization in Puthuperungulathur, Chennai, Tamil Nadu. All earthworms were kept in a moist soil mixed with cow dung. Distilled water was given to reach 60% of maximum water holding capacity. Soil was changed every four weeks and earthworms were maintained until required for experimentation. Five different type of waste was collected from different areas in and around Chennai The collected waste were chopped into small size and allow to decomposition for 15 days. The histology of gut of earthworm was studied adopting the routine paraffin method (Humason, 1979).Gut of earthworm, dissected out from the control and experimental animals, were blotted free of mucus, washed thoroughly in physiological saline, cut into pieces of desired size and fixed in Bouins fluid fixative immediately after autopsy. Fixation was carried out at room temperature for 24 hr, after which the tissues were transferred to 70% alcohol. Several changes of 70% alcohol were given until the yellow colour disappeared from the tissues. The tissues were then dehydrated by passing through ascending grades of alcohol, cleared in xylene, infiltrated with molten paraffin, and finally embedded in paraffin wax (58 C MP).Tissue section of 5-µm thick transverse and longitudinal sections were obtained using a rotary microtome (Leica, Germany). The sections, thus obtained, were stained in Harris hematoxylene and eosin, dehydrated using alcohol, cleared in xylene and mounted using dihydroxy phthalate xylol (DPX). The stained slides were observed in a CarlZeiss (Germany) Axio-2 Plus research microscope. RESULT AND DISCUSSION Histological studies in gut region of the earthworm E. eugeniae treated with different organic wastes a) control - No abnormalities detected b) Garbage waste- Increased goblet c) Sugarcane baggase - Pyknotic nucleus seen on gut epithelial d) Paper & Polythene Waste - Swollen gut epithelial and mucosal epithelial degeneration e) Textile dye sludge - Necrotic gut epithelial f) Herbicide - Pyknotic nucleus seen on gut epithelial Page 19

3 International Journal of Ethnomedicine and Pharmacological Research (2014), Vol.2 Issue 1, P Fig-1. Histological studies in gut region of the earthworm E. eugeniae treated with different organic wastes A B C D E F Fig-2. Histological studies in gut region of the earthworm E. fetida treated with different organic wastes A B C D E F Page 20

4 Histological studies in gut region of the earthworm E. fetida treated with different organic wastes a) Control - No abnormalities detected b) Garbage waste- Necrotic gut epithelial c) Sugarcane baggase - Necrotic gut epithelial d) Paper & Polythene Waste - Pyknotic nucleus seen on gut epithelial & increased goblet e) Textile dye sludge - Necrotic gut epithelial f) Herbicide Pyknotic nucleus seen on gut epithelial The observed pathological changes in the intestines were characterized by villous atrophy, enteritis with cellular infiltration and formation of characteristic granulomas. In contrast, there were no significant changes in the liver and kidneys. Such findings may be interpreted in the ground that the infection did not reach the liver and kidneys because the parasite cannot pass to the circulation. These observation and others were also reported by (Pinto et al.,2008) who demonstrated that the pathological changes in the intestines were characterized by villous atrophy, enteritis with cellular infiltration and formation of characteristic granulomas, and the same findings were confirmed by (Lalchhandama,2010). Cell death or necrosis was observed in this period. According to Bowen and Lochshin (1981) cell death is not a single entity but heterogenous structure, mechanism and biological function. Cell death or necrosis is characterized by pyknotic nuclei, cytoplasmic swelling and mitochondrial damage which results from failure in osmotic regulation caused by loss of cellular energy supplies. By the 40th day of exposure there was observed recovery of the epithelial lining. According to Stephenson (1930) recovery could be brought by the chloragogen. These are known to migrate to the wound or lost tissue and regenerate them. It is well known fact that earthworms have a great power of regeneration (Leblond and Walker, 1956 and Hammana et al., 1995). Conclusion The histology studies on earthworm the textile dye sludge showed higher damages in gut tissues. No abnormalities were observed in control category of earthworm. Textile dye sludge found to be having negative effect on the earthworm when compared to the other waste. REFFERENCE 1. Bowen, I D and Lockshin, R. A., In cell death in biology and pathology, pp London: Chapman and Hall. 2. Edwards C.A, and Fletcher K.E., Interactions between earthworms and microorganisms in organic matter breakdown, Agric. Ecosyst. Environ., 1988;24: Environment- the biotechnology of composing. - Advanced Biotech, 2: Garg V K, Kaushik P. Vermistabilization of textile mill sludge spiked with poultry droppings by an epigeic earthworm Eisenia foetida. Bioresour. Technol. 2005;96: Ismail S A., Joshi P. and Grace A. The waste in your dust bin is scaring theenvironment- the biotechnology of composing. Advanced Biotech. 2004;2: Joshi N. and Dabral M., Life cycle of earthworms Drawida nepalensis, Metaphire houleti and Perionyx excavatus under laboratory controlled conditions, Life science Journal, 2008;5: Kavitha V., Ramalingam, R. and Anandi, V., Effect of endosulfan on the bacterial and fungal populations in the gut of the Indian earthworm Lampito mauritii (Kinberg). J. Sci. Trans. Environ. Technov., 2008; 2( 2): Mall A.K., A. Dubey and S. Prasad. Vermicompost: An inevitable tool of organic farming for sustainable agriculture. - Agrobios Newsletter, 2005;3: Padma U., Rao R.S, and Srinivas N., Eco-friendly disposal of vegetable wastes. Fundamental Appl. Sci., 2002;1(1): Rao J V., Toxicity of Azodrin on the morphology and acetyl choliesterase Page 21

5 activity of the earthworm Eisenia fetida, Environ, Res., 2004;96: Reinecke S A. and Reinecke A J., The impact of organophosphate pesticides in orchards on earthworms in the western Cape, South Africa, Ecotoxicol. Environ. Safety, 2007;66: Stephenson J., The oligochaeta. Oxford Clarendon press.through vermitechnology. - J. Ecobiol. 1930:14: Townsend Lee and potter Dan (Entomology) and Powell A.J. (Agronomy), Earthworms: Thatch Busters, college of Agriculture, university of Ketucky, Vijaya, T M., Middha S K., Usha T, Saini D and G. Govindaraj, Morphological and Histological Studies on the Vermicomposting Indian Earthworm Eudrilus eugeniae, World Journal of Zoology, 2012;7(2): Yasmin S. and D souza D., Effect of pesticides on growth and reproduction of earthworm: A Review, Applied and Environmental soil science, 2010; Yasmin S. and D souza D., Effect of pesticides on the reproductive output of Eisenia foetida, Bull. Environ.Contam. Toxicol., 2007;79(5): Page 22

COMPARATIVE STUDIES ON THE LEVELS OF VITAMINS DURING VERMICOMPOSTING OF FRUIT WASTES BY EUDRILUS EUGENIAE AND EISENIA FETIDA

- 57 - COMPARATIVE STUDIES ON THE LEVELS OF VITAMINS DURING VERMICOMPOSTING OF FRUIT WASTES BY EUDRILUS EUGENIAE AND EISENIA FETIDA 1 M. LAKSHMI PRABHA, 2 INDIRA A. JAYRAAJ, 3 JEYARAAJ, R AND 1 SRINIVASA