Dr. Jeremy Van Boening - Field Perspective – Sample Handling & Collection


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Field Perspective – Sample Handling & Collection - Jeremy Van Boening, DVM, Republican Valley Animal Center P.C., Alma, NE; Anthony Smith, Ph.D, BioMed Diagnostics, from the 2014 NIAA Annual Conference titled 'The Precautionary Principle: How Agriculture Will Thrive', March 31 - April 2, 2014, Omaha, NE, USA.

More presentations at http://www.trufflemedia.com/agmedia/conference/2014_niaa_how_animal_agriculture_will_thrive

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  • Slide to Talk About Where we are and what we do.
  • Location to demonstrate our location close to the Kansas Border and the difficulties associated with moving breeding cattle back and forth. We also sent cattle or embryos to Kansas, Colorado, Texas, Ohio, Indiana, Missouri, Iowa, South Dakota, California, Montana, Utah, New Mexico, Wyoming, Oklahoma, as well as Canada, Australia, and New Zealand.
  • In 2008 Nebraska began to have Trich issues from a problem that we believe came from dumping infected bulls to salebarns in Nebraska from South Dakota’s outbreak west river. Cases in Nebraska increased for about 2 years. Nebraska enacted import rules. Nebraska Cattleman began discussing how to best protect our herds.
    Discovered the issue was very complicated:
    Commerece: Nebraska Salebarn Example
    Cows coming to Nebraska must be 4 months bred or have a calf at side or they cannot come into the state.
    Creates issues with a Kansas producer who wants to sell a the Nebraska sale barn because if they are bred under 4 months they must be sold for slaughter. Creates confusion!!!!!
    Near and Dear to those that had herds have trich or close proximity to trich
    Discussions from testing every bull upon every sale, all cull bulls must sold for slaughter, to neighbor notification.
    Discovered that a great deal of education was needed. Many producers have no idea what trich is.
    Discovered that legislation could impact many aspects of the industry.
    The different parts of the industry did not always agree with the other on proposed legislation
    Seedstock Producers Hate testing yearling bulls. One reported an 8% dropout of bulls attributed to testing.
  • Discoveries from Committee

    Many different opinions on testing methods.
    Many don’t believe testing is reliable or accurate.
    Most did not want regulation (statutory regs)
    Most believed the bad actors would circumvent any laws.
    Most wished all states would have similar regs.
    All wanted a better test.
  • # 1-5
  • I would use whichever method you are comfortable with and your D-lab accepts
  • I think we notice after culture on a dirty sample changes in the medium. It becomes less viscous and more fluid. What is killing the trich in here?
  • Some Labs will only do PCR from the in pouch. Having enough when a client calls with 200 head of yearlings to check.
  • Different responses from each lab of the 10 we surveyed.

    Very confusing for vets!!!!
  • Jeff Ondraks with Texam A and M Study needed.

    US Postal Services is NOT Reliable and in many cases not Timely.

    Need access to UPS or Fex Ex or other courier services.

    Survey Results: Confusion is created because labs can vary greatly on how they expect to receive samples.
    1. Results from 10 labs ranged from on ice, to one ice but not touching, to room temperature and ranged from needing the samples overnight up to 120 hours.
  • Dr. Jeremy Van Boening - Field Perspective – Sample Handling & Collection

    1. 1. Trichomoniasis Field Perspectives Jeremy VanBoening, DVM
    2. 2. Trichomoniasis Involvement • Clinical Practice • Nebraska Cattlemen-NCBA • Nebraska Veterinary Medical Association
    3. 3. Sample Collection Texas A&M Trich Sample Collection Video
    4. 4. Sample Collection Challenges Survey of 10 D-Labs 10-14-2013 Lab ID Do the samples need to be incubated? Do the samples need to be put on ice when shipped? How many samples can be pooled? Preferred collection medium? A Yes 24hrs Yes Up to 5 tubes B No No, dark and warm packaging Up to 5 InPouch C No, room temp No Up to 5, but not recommended InPouch D You can if you want, just state on submission form if you incubated samples or not. Have to receive samples within 72hrs of collection. If cold outside put a thawed ice pack. If hot outside put ice pack but not directly on samples Up to 5 InPouch or tubes E No, room temp, ship same day No No pooling InPouch F Yes or No, state on form if you did. Has to arrive at lab within 120hrs after collection. No Up to 5 InPouch G No cold pack Up to 5 Scraping in 5ml of Saline placed in tube. H No No Up to 5 InPouch I No No, room temp No pooling InPouch J No, ship overnight Yes Up to 3 wash in red top tube with saline
    5. 5. Summary of PCR Related Questions Incubation Yes= 2 No=6 Your Choice=2 Ship on Ice Yes=2 Room Temp=6 Other=2 Pooling Up to 5=7 No Pooling=2 Up to 3=1 Preferred Collection Media In Pouch=6 Saline=2 Tubes=2
    6. 6. Sample Collection Challenges Restraint and Safety
    7. 7. Is this safe??
    8. 8. What Collection Method 1. preputial scraping with pipette 2. preputial scraping using thrichit 3. saline rinse 4. swap or scraping of the penis NO BLOOD for Either PCR or Culture
    9. 9. Sample Collection Challenges Cleanliness  Field Collection Area  Scissors (Disinfect or Not and with what?)  Access to saline for rinsing
    10. 10. Dirty Samples • Rarely if Ever Return Positive • Recent Study Hustun, etal. MSU – 20 Samples w/ bacteria injected at increasing #’s – 20 Samples clean (Only T. Foetus Added) – 0 of 20 with bacteria ever recorded as positive – 20 of 20 w/out bacteria had at least one positive recording
    11. 11. Sample Collection Challenges Proper Collection Media
    12. 12. Sample Preparation  Depends on the D-Lab!!  Collection Medium (In-Pouch, Tubes, Saline)  Incubate or Not • Depends on D-Lab, PCR or Culture,  How should the sample be packaged? • On Ice, Room Temperature, Or Depends on the Weather
    13. 13. Transportation Issues • Rural Access to Timely Reliable Transportation • How to package and send samples? – How long can they be in transit and at what temperature? – Sensitivity and Specificity go down for both culture and PCR Hustun etal MSU
    14. 14. How Do We Get Better???? Standardization 1. Adopt the Best Diagnostic Testing Available (qPCR) Using Chemical-Lysis and IAC. 2. Promote SOP’s for Sampling, Sample Prep, and Shipping 3. Increased Veterinarian/Producer Awareness 4. Standardized Regulations