Abhishek Gupta.ppt

THREE MAIN PROBLEMS

HUGE AMOUNT OF POULTRY WASTE GENERATED EVERY YEAR

MOSQUITO BORNE DISEASES ARE MAJOR CAUSE OF MORTALITY IN TROPICAL COUNTRIES

HARMFUL EFFECT OF COMMERCIALLY AVAILABLE MOSQUITO REPELLENT

 According to Vogt 2006, 18million thousand tons of feather waste is generated all over the world.  Feathers are mainly composed of -keratin. Keratins are resistance to biodegradation.  Thus they are polluting the environment.

 Mosquito borne diseases such as malaria, filariasis, yellow fever and dengue are the major cause for morbidity and mortality and are a major economic burden within disease-endemic countries. According to Snow et al 2005, each year, about 300 million people are estimated to be affected by malaria, a major killer disease.  Such diseases are major threat to 2,400 million (about 40%) of the world’s population.  There are many cases of resistance development of mosquitoes towards chemical insecticides

Data Source: World Health Organization Map Production: Public Health Information and Geographic Information Systems (GIS)

Data Source: World Health Organization Map Production: Public Health Information and Geographic Information Systems (GIS)

 Mosquito repellent are composed of harmful chemical compounds like, • Pyrethrins • N,N-diethyl-meta-toluamide (DEET) • Permethrin • P-mentane-3,8-diol • Allethrin • Formaldehyde According to liu et al 2003, burning one mosquito coil would release the same amount of particulate matter mass as burning 75–137 cigarettes. The emission of formaldehyde from burning one coil can be as high as that released from burning 51 cigarettes.

COMPOUND

• EFFECTS

Pyrethrins

• Headache, Nausea and Dizziness

DEET

• Neurotoxin and Skin irritation

Permethrin

• Carcinogenic and Eye irritation

P-mentane-3,8diol

• Skin redness and Swelling

Allethrin

• Complications in pregnant women

Formaldehyde

• Irritation of upper respiratory tract

 Poultry wastes are mostly dumped near villages.  I went to Chittor for a survey of various poultry farms.  Poultry farms are near villages of Chittor.  Areas near the poultry farms were full of chicken feather waste.  Even the dead birds were thrown on which nearby fields, on which flies were breeding.  Whole scenario was looking full of disease.

The effluent from the factories and industries built near rural areas are the major breeding place for the mosquito.  Beside this many other terrains are present which support stagnant water, which allow mosquito breeding. This may lead to the spread of many mosquito borne diseases in the rural area, which has increased the rate of mortality.  People in villages are more likely to sleep on terrace and open ground, which exposes them to mosquito borne diseases to larger extent.

VILLAGE: JAGAT MAAJHA

 This survey was performed at village in U.P. India (14-27 June 2011) at: Gram: Jagat maajha Post: Karota District: Deoria  Survey was done for 10 houses.  All were with joint families with a mean of 14 members per house.  Around 8-10 members sleep on terrace or open ground.  They don’t use any commercial mosquito repellent but only use traditional techniques.  These includes burning of dried leaves or cow dung cakes to produce smoke, which are very irritating to them also.  Sarpanch told me that the budget allotted to them is not sufficient so as to use commercial methods like fumigation and mosquito repellent spray.  He also added that if any cheap method is developed they can invest money for the benefit of there villages.  This made me to develop an idea for developing cheap mosquito repellent.

 Feathers from poultry wastes are called Chicken Feather Wastes (CFW).  CFW are very cheap bioorganic waste, which can be used as a substrate for the production of mosquitocidal toxin.  -keratin present in feathers are resistance to degradation. But keratinase producing bacteria can be used to degrade them. One such bacterium is Bacillus sphaericus (Bs).  Bacillus sphaericus is an entomopathogenic, aerobic, rod-shaped, endospore forming gram positive soil bacterium.  Bs is effective against Culex spp. and Anopheles spp.  The main advantage of Bs is its low environmental toxicity and ability to overcome the development of resistance.

Collection of chicken feather waste from various poultry farm Washed, dried, powdered and stored at room temperature 10 gm powder + 1.5 litre water (pH 7.5) Autoclave the culture media Inoculated with pre culture entomopathogenic bacteria Kept in orbital shaker for constant agitation (till sporulation) Cell mass separated by centrifugation (10,000 rpm for 30 min at 4°C) Washing with NaCl solution and ddH2O

Cell biomass Sonication

1g Alumina

10µl Endotoxins

Limulus amoebocyte lysate React for 30 min @ 37°C

Gelation takes place

Estimation of Endotoxins by UV spectrophotometer (280 nm)

Limulus Amoebocyte Lysate (LAL) Assay Method

 A positive reaction is characterised by gel formation.  Whereas, in negative reaction the mixture remains liquid.  Distinction is made by tilting the plate at an angle, positive gel remains immovable whereas the negative drop flows.

 According to Zerdani et al 2004, feather waste is composed of protein (81%), fat (1.2%), ash (1.3%) and dry matter.  Also according to Suneetha et al, 2004, 2006 and 2010, feather consists of protein having a lot of sulphur containing amino acids (cysteine).  The feather waste is looked as a balanced medium. So, it does not need other nutrients for culturing the degrading bacteria (Bs).

CHICKEN FEATHER WASTE

81% PROTEIN

1.2% FAT

1.3% ASH

DRY MATTER

 The growth curve of entomopathogenic bacteria has 1 hr lag phase.  Rapid multiplication of bacterial cell was till 48 hrs.  After the cell lysis, spores containing endotoxins are released in medium and reaches to its peak concentration at 72 hrs.

 According to Poopathi and Abidha 2010, when the crystal toxins from Bs are ingested by mosquito larvae, and after solubilisation and proteolytic cleavage, the activated toxin interacts with the midgut epithelium leading to the death of larvae.  The receptor of this toxin is not available in larger animals including humans, thus they have no or very little ill effect on ecosystem.  Crystal toxin contain two major polypeptides called as BinA (42kDa) and BinB (51kDa).  BinB is responsible for binding to the mid gut receptor and the toxic effect is shown by BinA.

Raw Materials

• Free of cost. • Rs. 1000 per ton for transportation.

Manufacturing Cost

• Cost of production: Rs. 2000 per reactor. • Yield: 20 liters toxin per 10 kg CFW per reactor.

Packaging and Distribution

• Cost: Rs. 600 per 60 liters. • Will be distributed to Gram Sabha at villages.

Usage Cost per month

• Each liter will be diluted 1/10. • 30 liters of toxins will be sprayed in the village’s water bodies for 3 times a month. • Including all the costs Rs. 350 per month per village will spent excluding the equipment cost.

Mosquito Coil Rs. 25 for 10 coils i.e. Rs. 75 per month per house. But with it comes lots of harmful compounds.

Mosquito Spray Rs. 129 per 320 ml which last for one month per house. But with it comes lots of harmful compounds.

Cost of Commercial Mosquito Repellent AllOut liquid refill Rs. 81 for 90 nights + Rs. 50 machine i.e. approx Rs. 50 per month per house. But with it comes lots of harmful compounds

Good Night mat Rs. 60 for 30 mats + Rs. 20 machine i.e. approx Rs. 62 per month per house. But with it comes lots of harmful compounds

 Fumigation is generally used in urban areas to kill mosquitoes. But this kills other insects also like honey bee, ants, butterflies, etc. These innocent insects play a crucial role in ecosystem as pollinator, scavenger, etc. In urban areas there removal don’t have much effect but can create a great imbalance in ecosystem if used in rural areas. In villages the main source of income is agriculture and due to lack of pollinator there may be a great loss.

In villages the commercial repellent will not work in open areas and will have more severe ill effect when used indoor due to small rooms. Thus there is need of a mosquitocidal toxin which kills the roots of mosquitoes i.e. larvae. In India the total number of villages are 638596 (Census of India 2001). Due to lack of budget such a large numbers of villages are neglected and they need a cheap mosquito repellent. Since the toxins produced by us are safe and cheap it can be afforded by the Gram Sabha of villages. We will cost 7 thousand per 60 liters (sufficient for 60 spray for 20 month) of concentrated toxin with which a stainless steel knapsack sprayer will be provided free of cost to Gram Sabha.

 In India total number of villages estimated according to Census of India (2001) is 638596.  The number of families per villages ranges from 50 to 1630.  The cost of mosquito repellent used per family in rural area ranges from Rs. 15 to Rs. 25 per month  So if we estimate the cost spent on mosquito repellent per village the figure will be Rs. 12600 to Rs. 21000 per month.  Compared to this figure our mosquitocidal toxin is very cheap. For whole village it will only cost Rs. 350 per month. It is very much feasible in rural area. As it is very much eco friendly it can also be used in cities and town.

 Endotoxins from Bs provides effective alternatives to broad range of larvicides in many situations with little or no environmental impact.  Taking into account environmental benefits including safety for humans and other non-target organisms, reduction of pesticide residues in the aquatic environment, increased the activity of most other natural enemies and increased biodiversity in aquatic ecosystems.  It effectively reduces the population of mosquito, thus the overall consumption of harmful mosquito repellent is reduced. This reduces many human problems which arise from mosquito repellent.  Using mosquitocidal toxin produced from chicken feather waste (CFW) is very cost effective. The culture media preparation is cost free as it utilises the feather which is a waste.

TRIPLE BENEFITS TO VILLAGES

COMPLETE UTILIZATION OF CHICKEN FEATHER WASTE FROM VILLAGES

PRODUCTION OF MOSQUITOCIDAL TOXIN TO REDUCE MOSQUITO BORNE DISEASES IN VILLAGES

REDUCTION IN OVERALL CONSUMPTION OF HARMFUL MOSQUITO REPELLENT

1) V. Suneetha et al, Bioremediation of poultry waste, Advance biotech, Vol. 10, PP. 7-9, 2010 2) S. Poopathi, S. Abidha, International Biodeterioration & Biodegradation Vol. 62, PP. 479-482, 2008 3) V. Suneetha and Z. A. Khan, Soil Enzymology, Soil Biology, Springer-Verlag BerlinHeidelberg, PP. 259-269, 2011 4) S. Poopathi and S. Abidha, J. Physiology and Pathophysiology, Vol. 1(3), PP. 22-38, 2010 5) S. N. Hussanini and H. T. Hassanali, J. Med. Microbiology, Vol. 24, PP. 89-90, 1987 6) Liu et al, Environ. Health Perspectives, Vol. 111, No. 12, 2003