Since there are differences between sample types and sizes (e.g. chicken vs. turkey carcasses), be sure to check the specific program protocol before using this procedure. Note: BPW may be substituted for the Buffered Phosphate Diluent (BPD).
b. Aseptically transfer the carcass to a sterile Stomacher
equivalent), draining as much excess fluid as possible during the transfer.
Note: Larger (24 x 30-36 in.) bags will have to be used with turkeys.
c. Add 400 ml (chickens) or 600 ml (turkeys) of BPD to the carcass in the bag. Pour approximately one-half the volume into the interior cavity of the bird and the other half over the skin.
d. Rinse the bird, inside and out, with a rocking motion for 1 min at a rate of approximately 35 forward and back swings per minute. This is done by grasping the carcass in the bag with one hand and the closed top of the bag with the other. Rock with a reciprocal motion in an approximately one-foot arc, assuring that all surfaces (interior and exterior) are rinsed.
e. Aseptically remove the carcass from the bag, draining excess rinsed liquid into the bag, dispose of the carcass.
f. Pipette 10 ± 0.1 ml of the stomached 1:10 dilution into a 90 ml dilution blank to make a 1:100 dilution. Repeat this procedure to prepare serial dilutions of 10-3, 10-4, 10-5, and 10-6. More dilutions may be required if a sample is suspected of having a high microbial load > 107 CFU/g.
g. Shake all dilutions 25 times in an approximate a one-foot arc. Use a separate 10 ml pipette to prepare each dilution.
The analyst should strive to minimize the time from when the sample is stomached or blended until all the dilutions were placed in or on the appropriate medium; this time should not exceed 20 minutes.
Since there are differences between sample types and sizes (e.g. chicken vs. turkey carcasses), be sure to check the specific program protocol before using this procedure. Note: BPW may be substituted for the Buffered Phosphate Diluent (BPD).b. Aseptically transfer the carcass to a sterile Stomacherequivalent), draining as much excess fluid as possible during the transfer.Note: Larger (24 x 30-36 in.) bags will have to be used with turkeys.c. Add 400 ml (chickens) or 600 ml (turkeys) of BPD to the carcass in the bag. Pour approximately one-half the volume into the interior cavity of the bird and the other half over the skin.d. Rinse the bird, inside and out, with a rocking motion for 1 min at a rate of approximately 35 forward and back swings per minute. This is done by grasping the carcass in the bag with one hand and the closed top of the bag with the other. Rock with a reciprocal motion in an approximately one-foot arc, assuring that all surfaces (interior and exterior) are rinsed.e. Aseptically remove the carcass from the bag, draining excess rinsed liquid into the bag, dispose of the carcass.f. Pipette 10 ± 0.1 ml of the stomached 1:10 dilution into a 90 ml dilution blank to make a 1:100 dilution. Repeat this procedure to prepare serial dilutions of 10-3, 10-4, 10-5, and 10-6. More dilutions may be required if a sample is suspected of having a high microbial load > 107 CFU/g.กรัมจับโค้งหนึ่งเท้า dilutions ทั้งหมด 25 ครั้งในการประมาณ ใช้เปตต์ 10 ml แยกต่างหากเพื่อจัดเตรียมเจือจางแต่ละการวิเคราะห์ควรพยายามลดเวลาจากเมื่อตัวอย่าง stomached หรือผสมจนกว่า dilutions ทั้งหมดมีอยู่ใน หรือ บน สื่อที่เหมาะสม เวลานี้ไม่ควรเกิน 20 นาที
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