Animal Sciences Summer Internship 2004

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Animal SciencesSummer Internship 2004

• Department of Animal Sciences
• The Ohio State University
• Columbus, Ohio

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Background of Organization

• Dr. Eastridge works in the Department of Animal
  Sciences
• Focus of research in dairy cattle nutrition
• Coordinator of Animal Sciences Extension
• Professor and researcher
• Department of Animal Sciences
• Support Extension as resource for farmers
• Promote programs to educate youth in animal
  agriculture

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Responsibilities

• Journal Articles
• Twenty-five years of Journal of Dairy Science
  articles for review
• Separate articles into specific categories for
  organizational purposes

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Responsibilities

• NDF neutral detergent fiber
• Fatty acid
• Nitrogen determination

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Responsibilities

• Feeding research cows
• Collection of blood
• Cleaning glassware
• Ohio State Fair Youth skillathons

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Responsibilities

• Rumen Protected Choline Project
• Purpose to determine the effectiveness of three
  rumen protected choline products
• Why is choline important?
• Methyl donor which spares methionine
• Precursor to acetylcholine important for nerve
  synapses
• Helps to transport triglycerides from the liver
• BUT, choline is easily degraded in the rumen and
  so rumen protected products must be used to
  prevent deficiencies in this nutrient

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Rumen Protected Choline Project

• Establish a procedure
• AOAC method for choline determination in baby
  formula
• Made many equipment modifications
• Test assay on spiked samples and standard
  samples
• Determined percent recovery of assay
• Established limitations for concentration of
  choline to be assayed

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Rumen Protected Choline Project
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Rumen Protected Choline Project
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Rumen Protected Choline Project
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Rumen Protected Choline Project

• Preparation of in situ samples
• Three products to test
• Eight different time points
• Two samples for each product at each time point
• 48 samples placed in two different cows
• 5 grams weighed into dacron bags
• Sealed with heat sealer and tied with rubber band
• Placed on fish stringers and attached to metal
  chains
• Chains in laundry bags and placed in rumen

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Rumen Protected Choline Project

• Collection of in situ samples
• At each time point six total samples were taken
  from each cow
• SAMPLING FROM THE RUMEN
• Open the cannula.

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Rumen Protected Choline Project

• 2. Find the laundry bag with samples on the
  chains.

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Rumen Protected Choline Project

• 3. Remove samples from the bag.

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Rumen Protected Choline Project

• 4. Reseal the cannula.
• 5. Wash samples thoroughly with cold water.

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Rumen Protected Choline Project

• 6. Store in ziplock bags in freezer until
  analysis.

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Rumen Protected Choline Project

• Analysis of Choline Concentration from in situ
  samples
• Determine amount of subsample for concentration
  of 10 to 40 ug
• Weighed samples into tubes, mixed with 30 ml of
  HCl, and placed into a 70oC water bath for 3 hours

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Rumen Protected Choline Project

• 3. Adjust pH to 3.5 4.0
• 4. Centrifuge at 11,000 rpm for 5 minutes
• 5. Pour off supernatent and dilute to 50 ml

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Rumen Protected Choline Project

• 6. Filter and store filtrate in the refrigerator
• 7. Mix chromogenic reagent
• 8. Combine 0.1 ml of filtrate (or standard) with
  3 ml chromogenic reagent
• 9. Heat in 37oC water bath for 15 minutes
• 10. Dispense 0.3 ml of sample into 96-well plate
  and read at 505 nm in a spectrophotometer
• 11. Use equations to determine choline
  concentration

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Rumen Protected Choline Project

• Results
• Still interpreting results
• PhD student is running assay again
• Possible problems
• Subsample with concentration above limitations of
  assay?
• Assay not adequate to test these products?
• Error somewhere else in protocol overlooked?

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What Did I Learn in 10 Weeks?

• LOTS about dairy nutrition
• How to apply knowledge from classes
• Became more comfortable with Microsoft Excel,
  creating graphs, and data analysis

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What Did I Learn in 10 Weeks?

• Shed light on areas of strengths and weaknesses
• Strengths
• Working with people
• Enthusiasm for and understanding of science
• Weaknesses
• Quickly discouraged with new things
• Doubt my ability when run into roadblocks

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What Did I Learn in 10 Weeks?

• More about my career goals
• Probably not research
• Possible work in Extension or agricultural
  education
• Veterinary School still definite possibility

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Did I Value My Internship?

• YES!
• Diverse responsibilities made it interesting
• Met numerous intelligent people
• Saw various perspectives of a career in Animal
  Sciences which was a goal for this internship

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Any Improvements?

• Only one.
• Wish I had more knowledge about choline and the
  assay before I did the project
• Graduate students were more knowledgeable on the
  subject which led to frustration on my part

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ANY QUESTIONS?