Evaluation of Pyrus Pashia Leaf Extract As a Disinfectant for Rainbow Trout (Oncorhynchus Mykiss) Fertilised Eggs
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Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 697-706 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 1 (2017) pp. 697-706 Journal homepage: http://www.ijcmas.com Original Research Article http://dx.doi.org/10.20546/ijcmas.2017.601.084 Evaluation of Pyrus pashia Leaf Extract as a Disinfectant for Rainbow Trout (Oncorhynchus mykiss) Fertilised Eggs Ravindra1, Suresh Chandra2*, S.K.Mallik3 and R.S.Patiyal3 1ICAR-National Bureau of Fish Genetic Resources, Canal Ring Road, P.O.-Dilkusha, Telibagh, Lucknow-226002, Uttar Pradesh, India 2ICAR-Directorate of Coldwater Fisheries Research, Experimental Field Centre, Champawat, Uttarakhand, India 3ICAR-Directorate of Coldwater Fisheries Research Bhimtal-263136, Uttarakhand, India *Corresponding author ABSTRACT K e yw or ds In hilly areas, local population with their traditional knowledge utilize several medicinal herbs and plants parts for treating injuries and aliments of Rainbow trout, fertilised egg loss, their livestock, agricultural crop and human population. Among these Pyrus hatchery incubation, pashia, commonly known as Indian pear or Mehal is hardy wild mid hill disinfect ant, fruiting tree. Crude leave extract traditionally used for treating infection of leaf extract, Pyrus pashia toe finger particularly during rainy season. Fruit juice and ripe fruits of P. pashia used for treating eye injury and mouth sours. With a view to Article Info minimize use of chemical disinfectant in hatchery and to find out a locally Accepted: available suitable herbal substitute for disinfecting trout eggs, present 29 December 2016 comparative study was undertaken to observe the effect of leaves extract of Available Online: 10 January 2017 Pyrus pashia on egg survival and hatching during incubation phase. Introduction Among cultivable coldwater fishes, rainbow egg tripping, poor egg quality, physical injury trout, Oncorhynchus mykiss contributes handling and microbial infection greatly substantially in upland aquaculture production influence fertilisation and survival of eggs in India (Vass 2012). The species is widely during hatchery incubation. Exposure of trout preferred for farming owing to its hardy eggs to direct light or higher illumination for nature, tolerance to wide temperature longer period during incubation, fungal fluctuations and simple breeding protocol infections and frequent disturbance of eggs in (FAO 2012). Fertilised egg of rainbow trout rearing troughs may also reduce survival of takes a long incubation period of 3-14 weeks eggs and fry (Bell et al., 1971; Barker et al., for hatching under varied thermal regimes 1989; Griffiths 1991; Shepherd and Bromage (Woynarovich et al., 2011). Factors like lower 1992). Environmental parameters like sperm motility, over- ripening / untimely temperature, pH, dissolved Oxygen, ammonia 697 etc. also plays important role in spread of trout egg loss in hatchery (Heikkinen et al., infections and survival of eggs. Bacteria to 2013). At the same time, studies has also some extent are responsible for salmon egg revealed that ozone (Forneris et al., 2003), loss under a complex interaction between H2O2 (Gaikowski et al., 1998) and UV eggs, microbes and water quality parameters radiation (Heikkinen et al., 2013) may (Shepherd and Bromage 1992). During decrease egg hatching percentage during trout various developmental stages of eggs in eggs incubation. To achieve better egg hatchery, under unhygienic conditions, survival, antibiotics are also used. opportunistic and pathogenic microbes inhabit Unregulated use of chemicals and antibiotics over the eggs surfaces, thus, chances of egg in fish hatcheries and farms elicit harmful damage increases many folds (Barker et effects on fish and aquatic environment. al.,1991). Although, higher survival of trout Negative effect of several chemicals and eggs in hatchery could be achieved by antibiotics used in hatcheries are well known. selection of healthy brooders, maintaining With increasing awareness towards these optimum water quality parameters effects, safe methods are being explored to (Woynarovich et al., 2011). However, during minimize such losses and their side effects egg incubation phase, heavy loss of eggs has (Srinivasan et al., 2001). Many herbal extracts been reported due to microbial infections or paste for curing health disorders are now (Anon 2013). With growing rainbow trout widely being preferred and have been tried in farming in mid hill areas, demand for rainbow treating fish health disorders (Madhuri et al., trout fingerlings is also rising day by day. To 2012a; Pandey et al., 2012b; Dey and meet increasing seed demand, more numbers Chandra 1994, 1995; Ramasamy et al., 2011). of eggs are incubated in unit area, thus In hilly areas, local population with their chances of infection also increases several traditional knowledge utilize several times (Jeff et al., 2005). Improving water medicinal herbs and plants parts for treating quality supply of hatchery and maintaining injuries and aliments of their live stock, cleanness in egg rearing troughs through agricultural crop and human population regular removal of dead and decayed eggs, (Negi et al., 2011). These plants contain shells and other substrates help in reducing active ingredients which act as antimicrobial egg loss. However, in commercial trout seed and antifungal agents. Among these Pyrus production units, practically it is not always pashia (Buch-Ham ex D.Don), commonly possible to maintain all the desired parameters known as Indian pear or Mehal is hardy wild under optimum range. To maintain optimum mid hill fruiting tree belongs to family hygienic conditions, various anti fungal and Rosaceae and found in the mid Himalayan antibacterial chemicals have been tested in areas. The tree generally grows on sandy trout hatcheries with varying degree of loamy soil at a temperature range from -10 to protection. This includes malachite green, 35 0C. The bark leafs and fruits are formaldehyde, iodine, hydrogen peroxide, traditionally used both for consumption and bronpol and sodium chloride (Wanger et al., also as medicine (Jambey et al., 2012). Strong 2008). Malachite green was earlier considered antioxidant activity with free radical as one of the strong antifungal agent for scavenging activity has also been reported in treating fish eggs. However, malachite green flower extract of P. Pashia (Jianmin Hea, was banned worldwide in year 2002 owing of 2015). Crude leave extract traditionally used its harmful effects on animal health (Van for treating infection of toe finger particularly West 2006). Exposure of incoming hatchery during rainy season. Fruit juice and ripe fruits water, by U.V radiation significantly reduce of P. pashia used for treating eye injury and 698 mouth sours (Negi et al., 2011). With a view from naturally dried fallen leaves and above to minimize use of chemical disinfectant in methodology was followed for preparing hatchery and to find out a locally available extract of the dry leaves. Stock solution of suitable herbal substitute for disinfecting trout iodine named Betadine (Povidone iodine) eggs, present comparative study was containing 0.5% w/v iodine (Win Medicare) undertaken to observe the effect of leaves was procured from the local market and extract of P. pashia on egg survival and diluted to a concentration of 100mg/l. hatching during incubation phase. Rainbow Trout Egg Striping: Mature Materials and Methods healthy rainbow trout brooders with mean weight/length of 679.90±11.97g/371.00 Plant material: P. pashia is an angiosperm ±11.10mm were randomly collected from commonly known as Mehal or Mol in Hindi. brooder raceways and brought to hatchery for The tree is widely distributed in mid stripping. Approximately 35,000 nos. of eggs Himalayan areas in between 750-2600 msl were realized from 23 nos. of female rainbow and generally grows to a height of 3-6 m in trout brooders. The milt was poured over the sandy loamy soil. Although, small pear eggs and kept for 15 minutes in dark shaped fruits are edible but, they do not fetch conditions to accomplish adequate any market price and mostly consumed by fertilization. All batches of fertilized eggs wild animals. Locally crushed green leaves were mixed together to get uniformity of egg extract is used traditionally by hill people for stock. For different treatment, initially curing infections of toes. fertilized eggs were equally divided in four trays. Preparation of disinfectants: Newly budding reddish green leaves of P. pashia were Disinfection of trout eggs: Following collected from the trees located in fish farm of standard methods of egg disinfection, ICAR-DCFR Field Centre, Champawat in fertilized eggs were exposed in three Uttarakhand state of India, located at an treatments (Anon 2009). Before adding water altitude of 1620 msl. Collected leaves were in fertilized eggs, egg of first tray/trough were washed thoroughly in tape water and then exposed in 10% green leaves extract (GL) for allowed to dry under shade for 10 days. After 15 minutes, washed thoroughly in hatchery complete drying, leaves were grinded and 200 water and equally divided in three rearing g fine powder was made. The powder was troughs @ 2500±250/trough for further soaked in 800 ml 100% ethanol and placed in incubation. Fertilized eggs of second tray a shaking incubator for 24 hours. Soaked were similarly exposed in 10% prepared dry leaves powder in the above solvent was leaves extract (DL) for 15 minutes, washed filtered through 20µ mesh size