Standard Operating Procedures for Bull Management (1088.47 KB)

More documents

Recommendations

Info

v. Place the slide on the biotherm (warm stage at 37 degrees Celsius) of the phase contrast microscope and examine the motility (movement of individual sperms) at a magnification of 100/200 times (10X eyepiece and 10/20X objective). Descriptive assessment of individual motility- Very good 80-100% motile Good 60-79% motile Fair 40-59% motile Poor 40% motile Visual microscopic analysis of individual progressive motility is somewhat subjective even when performed by very skilled people and becomes tedious when large numbers of sample must be analyzed. Computer Assisted Semen Analysis systems have the potential to increase objectivity of analysis and reduce worker fatigue. w. Semen samples selected for freezing should have a minimum of 70% initial motility. x. Technique of Semen dilution- TWO STEP DILUTION AND PACKING AT 4° C Dilute the semen slowly with a small volume of warm Fraction A extender in a 50 ml dilution tube. Depending on the number of straws to be processed, the ejaculate could be than extended with Fraction A to a volume of 50 ml or less in the same 50 ml tube. The extended semen should be maintained at 28-32°C .Fraction A extender temperature and the semen temperature should be the same Cool the extended semen tube slowly to 4°C over a minimum of 2 hours by placing the tube inside a warm water bath (i.e. 250 ml –beaker with water at the same temperature as the extended semen), and then transferring the water bath to a cold handling cabinet or a cool room maintained at 4°C .Note: A 300ml volume of liquid at a temperature of 30-35°C will cool to 4°C over a period of 2 hours. After the cooling period, the extended semen is transferred to a flask that is precooled at 4°C At this time the remaining pre-cooled volume of Fraction A is added to the flask. The volume of Fraction A extender plus semen should be 50% of the planned final volume. The other half volume is Fraction B containing the glycerol that must be precooled at 4°C. Fraction B should be added gradually to the semen extended with Fraction A over a period of 30 minutes until a one to one ratio is reached. The final volume of extended semen will contain 7% glycerol concentration. The extended semen should be allowed to equilibrate at 4°C for at least 4 hours. During this time, the straws may be filled, sealed and placed on racks for freezing and counting
ONE STEP DILUTION Match extender temperature to semen temperature and maintain them in water bath, usually this is at 28-32°C . Dilute the semen slowly with a small volume of extender in a 50 ml dilution tube. Depending on the number of straws to be processed, extend the ejaculate to a volume of 50ml or less in the same 50ml tube. Cool the extended semen tube slowly to 4°C over a minimum of 2 hours by placing the tube inside a warm water bath (i.e. 250 ml –beaker with water at the same temperature as the extended semen), and then transferring the water bath to a cold handling cabinet or a cool room maintained at 4°C .Note: A 300ml volume of liquid at a temperature of 30-35°C will cool to 4°C over a period of 2 hours. After the cooling period, the extended semen is transferred to a flask that is precooled at 4°C Add the remaining amount of pre-cooled extender required to extend the ejaculate to the final processing volume. After the initial cooling period, the extended semen should be allowed to equilibrate at 4°C for at least 4 hours. During this time, the straws may be filled, sealed and placed on racks for freezing and counting ONE STEP DILUTION AND PACKING AT ROOM TEMPERATURE (20°C) Match extender temperature to semen temperature and maintain them in water bath, usually this is at 34°C. Dilute the semen slowly with a small volume of extender (1:1) in a collection tube. Do the final dilution by adding the pre-diluted semen to the extender. Leave the flasks of diluted semen at room temperature (20°C) for 10-15 minutes. Pack the semen at 20°C and placed it on racks Semen straws should be allowed to equilibrate at 4°C for at least 4 hours in cold handling cabinet . y, Dilution and packing work should be done under a LAFU. z.All used glass wares are immersed in a tub containing a neutral detergent to prevent drying and ease in cleaning. Printing, filling and sealing of straws a. Sterilized French mini straws (0.25 cc) of reputed companies are used. b. Printings of straws is done either before filling or after filling by use of an automatic straw printing machine / Jet printer and the straws are imprinted with the details of bull number / name / breed / name of the organization / batch number / year etc. c. The clarity of printing on all straws should be checked. d. Unused straws shall be repacked air tightly under the LAFU and sealed in a sealing unit for future use.
Page 1 and 2: TABLE OF CONTENTS Sl.No. C h a p t
Page 3 and 4: Standard Operating Procedures for B
Page 5 and 6: Except for very tender and succulen
Page 7 and 8: In places where the individual bull
Page 9 and 10: Preparation of bulls for collection
Page 11 and 12: Hygiene and Precautions required by
Page 13 and 14: g. Hold the AV before and after eac
Page 15 and 16: . Standard Operating Procedures (SO
Page 17: d. No staff from the semen collecti
Page 21 and 22: d. A ply wood/acrylic sheet is plac
Page 23 and 24: Chapter - 4 Standard Operating Proc
Page 25 and 26: Definition Description 0- None No m
Page 27 and 28: B. Wet technique From the thawed fr
Page 29 and 30: -B- 0.25 ml of frozen thawed semen
Page 31 and 32: Stock solution B 1. Nacl -9.01 gm A
Page 33 and 34: Volume) inactivated and sterilized
Page 35 and 36: 2. Water bath capable of being main
Page 37 and 38: SELECTING A SOLUTION FOR DILUTION:
Page 39 and 40: Area of 80 small squares = 1/400X80
Page 41 and 42: -Prepare semen smear and allow it t
Page 43 and 44: 6-cleaning and drying After equilib
Page 45 and 46: Sl. No Date of completion of Quaran
Page 47 and 48: Annexure-5 Q.C. test conducted duri

Standard Operating Procedures for Bull Management (1088.47 KB)

Create successful ePaper yourself

Delete template?

Save as template?