Title of Invention

AN IMPROVED FORMULATION OF NUCLEAR POLYHEDROSIS VIRUS AND A PROCESS FOR PREPARING THE SAME

Abstract

The present invention relates to a process for preparing a novel formulation of a Nuclear Polyhedrosis Virus (NPV) and for such formulations. The process comprises of adding and mixing NPV with a catalyst which also acts as stabilizer such as 0.01% to 4% sulphide, 0.1% to 5% chloride, 0.1% to 5% thiosulphate of sodium or potassium in water in the presence or absence of isopropyl alcohol or propylene glycol or ethylene glycol, the pH in the range of 4-9 at ambient temperature and the resultant solution is then checked for the POB (polyhedral occlusion body) count at the end of the 13 months which should be in the range of 0.5 x 108 to 2 x 108 and then packed in plastic bottle of suitable size for storage at room temperature.

Full Text

FORM 2 THE PATENT ACT 1970 & The Patents Rules, 2003 COMPLETE SPECIFICATION (See section 10 and rule 13) 1. TITLE OF THE INVENTION "AN IMPROVED FORMULATION OF NUCLEAR POLYHEDROSIS VIRUS AND A PROCESS FOR PREPARING THE SAME" 2. APPLICANT : (a) NAME SUDARSHAN CHEMICAL INDUSTRIES LTD. (b) NATIONALITY : An Indian Company registered under the provisions of the Companies Act, 1956 (c) ADDRESS 162, Wellesley Road, PUNE-411001. Maharashtra State, India 3. PREAMBLE TO THE DESCRIPTION PROVISIONAL The following specification describes the invention. 4. DESCRIPTION (starts from page 2) COMPLETE The following specification particularly describes the invention and the manner in which it is to be performed. 5. CLAIMS : Given on separate page. 6. DATE AND SIGNATURE : Given at the end of last page of specification. 7. ABSTRACT OF THE INVENTION : Given on separate page. This invention relates to a process of development of a novel clear odour less formulation of Nuclear Polyhedrosis Virus (NPV) product. This is normally used as a biocontrol agent in agriculture for two major pests namely Helicoverpa armigera & Spodoptera litura (Two pest larvae). They can attack various crops like cotton, soybean, tomato, chickpea, green pea & several others. These pests are widely distributed in India and attacks variety of cultivated & wild plants through out its distribution range. The increased chemical resistance to these pests is well known and need for biological control product has been felt. Prior Art: There are few biocontrol products for control of larvae of Helicoverpa armigera and Spodoptera litura. Use of Nuclear Polyhedrosis Virus has been tried. It is a very effective microbial biopesticide. However there are following disadvantages with the currently available formulations. 1. It needs to be stored at low temperature in refrigerator. 2. The product normally becomes turbid after few weeks and normally has very foul smell 3. The efficacy of the product also goes down after prolong storage of 6-8 months at room temperature. 4. It is not possible to store product at refrigerator at many locations (all over India) Therefore, according to the most important object of this invention is to aim to provide a effective, clear and odourless formulation of NPV which can be stored at room temperature for more than a year and have improved efficiency. It is another object of this invention to have a good quality of bioproduct formulation having a higher shelf life and higher polyhedral occlusion body (POB) count leading to improved results and providing a process for preparing the same. According to this invention, a process of development of NPV product formulation for Helicoverpa & Spodoptera pest control comprises treatment of virus solution containing a catalyst which also works as a stabilizer such as sulphide, chloride, azide, thiosulphates of sodium & potassium in water and in presence or absence of isopropyl alcohol, propylene glycol, ethylene glycol etc. They are used in pH range from 4-9 at ambient temperature. According to the present invention, therefore, a process for preparing an improved formulation of a nuclear polyhedrosis virus (NPV) comprises adding and mixing this NPV with a catalyst which also acts as stabilizer which is a combination of 0.01% to 4% sulphide, 0.1% to 5% chloride, 0.1% to 5% of azide, 0.1% to 5% thiosulphate of sodium or potassium in water in the presence or absence of isopropyl alcohol or propylene glycol or ethylene glycol, which if present are in the range of 0.01% to 0.4%, having pH in the range of 4-9 at ambient temperature and the resultant solution is then checked for the Q POB (polyhedral occlusion body) count which should be in the range of 0.5 x 10 to 3 x 109at the end of 13 months and then packed in plastic bottle of suitable size for storage at room temperature. According to further novel feature, the presence of catalyst cum stabilizer increases the efficiency of the formulation. The process is further illustrated with the examples as under. The procedure used for the production involve - 1) Larval rearing of Helicoverpa armigera & Spodoptera litura on semi synthetic diet. 2) Production of the virus on the same. It was done as mentioned below : NPV PRODUCTION PROCESS - The process can be illustrated as under. The semi synthetic diet is prepared. It contains chickpea flour, vitamins, sugars, oils and water (all mixed to blend with). The larvae hatched from the eggs are spread on the diet plate. After completion of 2 to 3 instars of the larvae they were then put on to the new diet, which is already contaminated with virus culture. This process is called diet contamination method. The virus is applied on the surface of the artificial diet. The same is used as the larvae feed. The virus then enters in to the digestive track of the pest larvae through feed. It then multiplies in the body. Due to this the metabolism of the larvae is disturbed and it stop feeding and then dies within 3-4 days period. The virus till then continuously multiply in the body of the larvae. The virus infected dead larvae from the larvae tray are then harvested after 6-8 days of infection and subsequently macerated in mixer / blender with small quantity of sterile distilled water (Sample A). The solution is then diluted with sterile distilled water to adjust the virus count (POB per ml.) to 10-8,10-9 numbers/ml (Sample B). The one larval extract will give us 0.5 to 2 ml of concentrated product which is further formulated as given below: Process of Formulation of the product - The formulation of the product (Sample B) was then done as follows. In the diluted product the catalyst cum stabilizer was then added and mixed properly. The catalyst cum stabilizer that were added include combination of sulfide, chloride, azide, thiosulphate of sodium or potassium in water and in presence or absence of isopropyl alcohol, propylene glycol or ethylene glycol. The pH varies from 4-9. Further this solution is checked for the POB count and then packed in the plastic bottles of suitable size. Example I The sample B is used as material to be formulated in this example. It has a pH of 6.5 & contains POB 3 x 108 No/ml. To this then is added 1 gm/L of sodium sulphide, 10 gm/L KCL, 10 ml propylene glycol & 5 ml isopropyl alcohol per litre. The formulation was then stored at room temperature for a period of more than one year alongwith experimental control. Sample B itself was used as control. The POB count at the end of o 13 months was 2 x 10 No/ml. Count in experimental control was 1x108 No/ml. Also control sample has got foul smell and was highly turbid. The Nuclear Polyhidrosis Virus laboratory bio assay was then conducted on the above formulations in example No.l. The formulation as above mentioned showed 60-85% mortality of the larvae on 8th day after treatment and has got no smell at all. The experimental control sample showed 0 to 55% mortality of the larvae on the 8th day. It also had very foul & decaying stnell. Example II In this experiment the sample B was used as materia/ to 6e formulated. ft has a pH of 7.5 & contains POB 3 x 108 No/ml. To this then is added 10 gm/L sodium sulphide & 5 ml/L of propylene glycol. Rest of the conditions were same as experiment I. The POB count at the end of 13 months was 1 x 108 No/ml. Count in experimental control was also 0.5 x 108 No/ml. Also control sample has got foul smell and was highly turbid. The Nuclear Polyhedrosis Virus laboratory bio assay was then conducted on the above formulation. The formulation as above mentioned showed 60-65% mortality of the larvae on 8th day after treatment and has got no smell at all. The experimental control sample (i.e. sample B) showed 0 to 55% mortality of the larvae on the 8th day. It also had very foul & decaying smell. Example III In this example all the conditions are same as in experiment I, except that here 10 gm/L of sodium azide, 2 gm/L of sodium chloride and sodium thiosulphate & 10 ml/L of propylene glycol were used. The remaining procedure followed is same as mentioned in Example no. I. The formulation was then stored at room temperature for a period of more than one year along with experimental control. Sample B itself was used as control. The POB count at the end of 13 months was 2 x 108 No/ml. Count in experimental control was also 0.5 x 108 No/ml. Also the control sample has got foul smell and was highly turbid. The Nuclear Polyhedrosis Virus laboratory bio assay was then conducted on the above formulations. The formulation as above mentioned showed 60 -85% mortality of the larvae on 8 day after treatment and has got no smell at all. The experimental control sample showed 0-55% mortality of the larvae on the 8th day. It also had very foul & decaying smell. Example IV The sample B is used as material to be formulated in this example. It has a pH of 5.9 & contains POB 3 x 108 No/ml. To this then is added 5 gm/L sodium azide, 1 gm /L sodium sulphide, 10 gm/L KCL, 10 ml/L propylene glycol & 5 ml/ L isopropyl alcohol. The formulation was then stored at room temperature for a period of more than one year along with experimental control. Sample B itself was used as control. The POB count at the end of 13 months was 2x108 No/ml. Count in experimental control was 1x108 ml/L Also control sample has got foul smell and was highly turbid. The Nuclear Polyhedrosis Virus laboratory bio assay was then conducted on the above formulations. The formulation as above mentioned showed 60-62% mortality of the larvae on 8th day after treatment and has got no smell at all. The untreated control sample showed 0 to 55% mortality of the larvae on the 8th day. It also had very foul & decaying smell. The formulation Nuclear Polyhedrosis Virus with catalyst cum stabilizer of the invention can be stored at room temperature more than a year without significantly loosing its efficacy. Further the characteristic in that the formulation developed remains clear and/ or odourless for a period of more than one year. WE CLAIM : 1. A process for preparing a novel formulation of a Nuclear Polyhedrosis Virus (NPV) comprises adding and mixing this NPV with a catalyst which also acts as stabilizer such as 0.01% to 4% sulphide, 0.1% to 5% chloride, 0.1% to 5% thiosulphate of sodium or potassium in water in the presence or absence of isopropyl alcohol or propylene glycol or ethylene glycol, the pH in the range of 4-9 at ambient temperature and the resultant solution is then checked for the POB (polyhedral occlusion body) count at the end of the 13 months which is in the range of 0.5 x 108 to 2 x 108 and then packed in plastic bottle of suitable size for storage at room temperature. 2. The process as claimed in claim 1 wherein the catalyst stabilizer is a combination of 0.01% - 3% sodium sulphide, 0.1% - 5% KCL, 0.1% - 5% propylene glycol and 0.1% - 5% isopropyl alcohol per litre. 3. The process as claimed in claim 1 wherein combination of 0.1% - 5% sodium azide, 0.1 to 5% sodium chloride and 0.01% - 5% propylene glycol as catalyst cum stabilizer are used. 4. An formulation containing a Nuclear Polyhedrosis Virus (NPV) with catalyst cum stabilizer as described in Claim 1. 5. A process for preparing a formulation as claimed in claim 1 substantially as herein described with reference to the examples in the specification. Dated this 08th Day of March, 2005 M.D.BHATE AGENT FOR APPLICANT

Documents:

310-mum-2005-abstract(14-2-2006).doc

310-mum-2005-abstract(14-2-2006).pdf

310-mum-2005-abstract.doc

310-mum-2005-abstract.pdf

310-mum-2005-cancelled pages(14-2-2006).pdf

310-mum-2005-claims(granted)-(14-2-2006).doc

310-mum-2005-claims(granted)-(14-2-2006).pdf

310-mum-2005-claims.doc

310-mum-2005-claims.pdf

310-mum-2005-correspondence(14-2-2006).pdf

310-mum-2005-correspondence(ipo)-(25-1-2006).pdf

310-mum-2005-correspondence-received-08042005.pdf

310-mum-2005-correspondence-received-14022006.pdf

310-mum-2005-correspondence-received.pdf

310-mum-2005-description (complete).pdf

310-mum-2005-form 1(22-3-2005).pdf

310-mum-2005-form 18(10-11-2005).pdf

310-mum-2005-form 2(granted)-(14-2-2006).doc

310-mum-2005-form 2(granted)-(14-2-2006).pdf

310-mum-2005-form 9(12-4-2005).pdf

310-mum-2005-form-1.pdf

310-mum-2005-form-18.pdf

310-mum-2005-form-2.doc

310-mum-2005-form-2.pdf

310-mum-2005-form-26.pdf

310-mum-2005-form-9.pdf

310-mum-2005-power of authority(28-2-2005).pdf