Title of Invention

A METHOD FOR PREVENTION OF DIAPAUSE IN BIVOLTINE SILKWORM EGGS BEFORE EGG LAYING

Abstract

"A method for preventing diapause in bivoltine silkworm eggs before egg laying" This invention relates to a method for preventing diapause in bivoltine silkworm eggs before laying of eggs comprising in mating the bivoltine females with males of a specially developed silkworm race KS-10, wherein the pupae of this race is subjected to a specific temperature of 23-25°C and humidity of 75-80% for 6-8 days. Dated this 4th day of September 2006 BRIEF ABSTRACT OF INVENTION Silkworm {Bombyx mori L.) races are classified on the basis of voltinism as Univoltines, Bivoltines and Multivoltines (or polyvoltines), depending upon the number of generations possible in a year, under natural conditions. In tropics generally, all the silkworms producing diapausing type of eggs are referred to as bivoltines and the non-diapausing as multivoltine or polyvoltines. Generally, polyvoltine eggs under tropical conditions hatch in 10 to 11 days of laying whereas in bivoltine silkworms do not hatch. They are artificially hatched by acid treatment or low temperature hibernation schedule. Acid treatment is the process in which, bivoltine silkworm eggs are treated with Hydrochloric acid of a particular specific gravity (ranging from 1.076 to 1.1) at a definite temperature (ranging from 25 to 48.6°C) for a specific period of time (ranging from 5 to 90 minutes). KSSRDI has developed a novel method of hatching bivoltine eggs alternate to the artificial hatching currently in vogue.

Full Text

A method for preventing diapause in bivoltine silkworm eggs before egg laying. Field of Invention: This invention relates to a method of preventing diapause in bivoltine silkworm eggs before laying of eggs. Background of the invention: Under natural conditions, in the diapausing type of eggs the embryo grows to a particular stage and enters diapause. Hence in silkworms, it is referred to as embryonic diapause. Once the embryo enters diapause, it hibernates through a specific period of time in low temperature. When it experiences favourable temperature, it resumes its growth and then hatches. However, after the eggs are laid, if they are subjected to an artificial treatment at appropriate embryonic developmental stage, it is possible to stimulate further growth of the embryo. This artificial treatment of the embryo leading to its hatching is called Artificial Hatching. Many methods were found partially or completely effective in artificial hatching. In physical method, temperature, friction, hot water treatment, air pressure, electric stimulus, ultra violet treatment, supersonic treatment, etc have been attempted. In chemical treatment sulphuric acid, nitric acid, hydrochloric acid (HCI), enzyme, ozone, perchloric acid treatment have been attempted. Among these, hydrochloric acid treatment is found to be of economic and of practical importance and referred to as "Acid Treatment". Though the mechanism of prevention of diapause through HC1 is yet to be understood, the technique is standardized. At present acid treatment is applied to eggs before they enter diapause. This requires expertise, precision and proper equipment. There are two methods of acid treatment. 1) Hot acid treatment: In this method 15-22 hours old eggs are treated with hydrochloric acid of 1.076 specific gravity for 5-6 minutes, at46.1°C. 2) Cold acid treatment: Here, 15-22 hour old eggs are treated with hydrochloric acid of 1.1 specific gravity for 35-90 minutes, at room temperature. Short term chilled eggs are also treated in this method. In physical method, diapausing silkworm eggs are subjected to low temperature, similar to the prevailing conditions of temperature regions. In India, this method is standardized under five categories and termed as "Hibernation method". 1) Short term Chilling: 30-35 hour old eggs are subjected to low temperature of 5°C and relative humidity (RH) of 70-80% for 40-70 days. After short term chilling eggs are cold acid treated. 2) Ordinary Chilling: 42-50 hour old eggs are subjected to low temperature of 5°C and RH of 70-80% for 60-120 days. After chilling, eggs are cold acid treated. 3) 6 Months hibernation schedule: 10-20 days old eggs (during this period they are kept at 25°C with 75-80% RH for aestivation) are subjected to low temperature of 5°C and relative humidity of 70-80% for six months. 4) 9 Months hibernation schedule: 20-30 days aestivated eggs are subjected to low temperature of 5°C and RH of 70-80% of nine months. 5) 11 Months hibernation schedule: 30-50 days aestivated eggs are subjected to low temperature of 5°C and 2.5°C and RH of 70-80% for eleven months. In each of the schedules while the eggs are brought to low temperature, they are subjected to intermediary temperatures of 20°, 15° and 10°C for a definite period. Similarly at the time of release the eggs are subjected to 10°, 15° and 20°C for a specified period before acid treatment and/or incubation. In absence of any other effective methods, either acid treatment or hibernation method is followed. Many studies have indicated that dipping of eggs in acid for considerable duration of time exerts stress on the developing embryo and weakens it. Acid treatment requires infra structure such as acid treatment bath, room heaters, hydrometer, thermometer, acid washing chambers, skilled man power, etc. Disposed waste acid causes pollution. Hibernation method requires huge infrastructure of cold storage units with facilities for maintaining required temperature and humidity with power back up facilities. During cold storage certain degree of damage could be caused to the silkworm eggs, because of human error or duration factor or power fluctuation. As a result, blue dead eggs, dried eggs, white floppy eggs and irregularly hatched eggs are often noticed. Objects of the Invention: An object of this invention is to propose a method for prevention of diapause in bivoltine silkworm eggs before they are laid. Another object of this invention is to propose a method for prevention of diapause in bivoltine silkworm eggs before such eggs are laid wherein the eggs need not be handled for diapause prevention process after they are laid. Yet another object of this invention is to propose a method for prevention of diapause in bivoltine silkworm eggs before such eggs are laid to avoid damaging of eggs due to handling and acid burning. Further object of this invention is to propose a method for prevention of diapause in bivoltine silkworm eggs before they are laid to avoid formation of damaged eggs due to error in the cold storage treatment. Another object is to propose a method for prevention of diapause in bivoltine silkworm eggs before they are laid which is easy, convenient, ecofriendly and economically advantageous. Further objects and advantages of this invention will be more apparent from the ensuing description. Detailed Description of Invention: In accordance with the invention, prevention of the diapause is effected before the egg laying by the mother moth unlike in alternative methods, where bivoltine eggs are processed for prevention or termination of diapause after they are laid. The method comprises in mating the bivoltine females with males of a specially developed silkworm race KS-10. The pupae of this race is subjected to a temperature and humidity treatment. The males emerge from such processed pupae of the race, KS-10 has a unique ability to convert bivoltines into polyvoltines. These males when crossed with bivoltine females, triggers the process of hatching in the eggs laid by these bivoltine females, as a natural phenomenon without acid treatment or any other diapause breaking treatment. Fig. 1: The actual sample of hatched shell is shown in the egg card. The females used in this case is a bivoltine race called NB4D2. The males are of KS-10. The eggs have been hatched without any treatment of acid or cold storage. Fig, 2: The actual sample of hatched eggshell shown in the egg card. The female used in this case is a bivoltine race called CSR2. The males are of KS-10. The eggs have been hatched without any treatment of acid or cold storage. Processing of Pupae: The pupae of this race is subjected to a specific temperature and relative humidity regimen. Male and female pupae can be identified morphologically by naked eye. The male pupae of KS-10 are handpicked and kept inside an incubator. The incubator temperature is preconditioned for 23-25°C and humidity for 75-80% (Relative humidity). The pupae are thus processed for 6-8 days. Later they are removed and kept in trays under normal temperature and humidity conditions. After this process the moths will emerge in another 3-4 days. Such processed males are mated with any bivoltine female. Steps involved in Crossing: Bivoltine females as well as KS-10 males are hand picked and put together so as to facilitate mating between them. They are placed inside a ring called 'Cellule" for about 3-4 hours for completion of mating. By this time the sperms are transferred from males to the 'Spermathecae* of female. The females are then separated from males manually and kept on a 'egg-card' for egg laying. The eggs laid by these bivoltine females hatch under normal conditions in about 10-11 days without requiring acid treatment or cold treatment. We claim: 1. A method for preventing diapause in bivoltine silkworn eggs before laying of eggs comprising in mating th 2. The mating as claimed in claim 1, wherein the KS-10 males and bivoltine females are placed inside a ring "Cellule" for about 3-4 hours for completion of mating. 3. The pupae of KS-10 as claimed in claim 1 are kept inside a preconditioned incubator having temperature of 23 to 25°C and humidity of 75-80%. 4. The processing of pupae as claimed in claim 1 wherein they are removed after the treatment to normal temperature and humidity conditions for 3-4 days. 5. The specially developed silkworm race KS-10 as claimed in claim 1, wherein they mate with any bivoltine females triggers the process of hatching the eggs laid by these bivoltine females. 6. The triggering of hatching as claimed in claim 5 is a natural phenomenon without any diapause breaking treatment. Dated this 4th day of September 2006

Documents:

1263-che-2005 complete specification as granted.pdf

1263-che-2005 correspondance others.pdf

1263-che-2005 abstract-12-08-2009.pdf

1263-che-2005 claims-12-08-2009.pdf

1263-che-2005 correspondence others-12-08-2009.pdf

1263-che-2005 description (complete)-12-08-2009.pdf

1263-che-2005-abstract.pdf

1263-che-2005-claims.pdf

1263-che-2005-correspondnece-others.pdf

1263-che-2005-description(complete).pdf

1263-che-2005-description(provisional).pdf

1263-che-2005-drawings.pdf

1263-che-2005-form 1.pdf

1263-che-2005-form 5.pdf

EXAMINATION REPORT REPLY.PDF