Larvae Lab Report
Results
Larval Studies
Normal Honey Bee Development of the Control Plate By day two, it seemed that three of the larvae did not survive due to their small size compared to the others. By day four, it seemed that four did not survive; however, movement was present in the others. By day eight, most of the larvae died due to mold development. However, three of the larvae started to progress into the pupal stage, but some of them did not complete the transformation. At day eleven, there were signs of eye development present in the larvae who reached the pupal stage. By day thirteen it was determined that eight out of the twenty-four survived, but only three out of the eight made it to the pupal stage. At day sixteen, the body was beginning to …show more content…
Those colonies were subjected to a Gram-stain to ensure that P. larvae, a Gram-positive bacterium was isolated (Figure 7). However, there was a presence of other bacteria visible on the Gram-stain. Therefore, the catalase negative colonies were streaked onto MYPGP agar to further isolate P. larvae. The pale colonies that grew on those plates were subjected to a 3% potassium hydroxide test. No signs of cell lysis was present; therefore, P. larvae was isolated.
Isolating DNA from the Pure Colonies Grown In addition, DNA were isolated from the colonies grown and its concentration was 33.6 ng/uL and the A260/280 value was 1.84. Next, an agarose gel electrophoresis was conducted to see if any DNA fragments were present. A band was visible and that validates that genomic DNA has been isolated.
PCR Amplification to Determine Strain of P. larvae with ERIC-Specific Primers ERIC-specific primers were used to genotype the genomic DNA of P. larvae isolated from the spore solution. A faint band present at 970 bp was present within all six lanes. Also, a prominent band was migrating around 1200 bp within all six lanes. Additionally, a double band between 1500 bp and 2000 bp can be seen in each lane. This was compared to an ERICI positive control to further validate the results (Figure
Larval Studies
Normal Honey Bee Development of the Control Plate By day two, it seemed that three of the larvae did not survive due to their small size compared to the others. By day four, it seemed that four did not survive; however, movement was present in the others. By day eight, most of the larvae died due to mold development. However, three of the larvae started to progress into the pupal stage, but some of them did not complete the transformation. At day eleven, there were signs of eye development present in the larvae who reached the pupal stage. By day thirteen it was determined that eight out of the twenty-four survived, but only three out of the eight made it to the pupal stage. At day sixteen, the body was beginning to …show more content…
Those colonies were subjected to a Gram-stain to ensure that P. larvae, a Gram-positive bacterium was isolated (Figure 7). However, there was a presence of other bacteria visible on the Gram-stain. Therefore, the catalase negative colonies were streaked onto MYPGP agar to further isolate P. larvae. The pale colonies that grew on those plates were subjected to a 3% potassium hydroxide test. No signs of cell lysis was present; therefore, P. larvae was isolated.
Isolating DNA from the Pure Colonies Grown In addition, DNA were isolated from the colonies grown and its concentration was 33.6 ng/uL and the A260/280 value was 1.84. Next, an agarose gel electrophoresis was conducted to see if any DNA fragments were present. A band was visible and that validates that genomic DNA has been isolated.
PCR Amplification to Determine Strain of P. larvae with ERIC-Specific Primers ERIC-specific primers were used to genotype the genomic DNA of P. larvae isolated from the spore solution. A faint band present at 970 bp was present within all six lanes. Also, a prominent band was migrating around 1200 bp within all six lanes. Additionally, a double band between 1500 bp and 2000 bp can be seen in each lane. This was compared to an ERICI positive control to further validate the results (Figure