SC Pullorum Typhoid Plate Testing & Banding Procedures (aka Pullorum Testing & Banding for Dummies)

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1 (for South Carolina Dr. Julie Helm, , 1. Items needed: a. SC NPIP wing or leg bands with pliers b. Pullorum Typhoid antigen c. Test plate d. Needles (22, 20 gauge) with syringe e. Collection loop f. Cup of water g. Paper towel h. SC NPIP Pullorum Typhoid Testing Report 2. Band the bird. a. All birds that are Pullorum tested must be identified with a SC NPIP numbered leg or wing band and recorded on the official Pullorum Typhoid Testing Report. If the bird already has an official state ID band from SC or any other state NPIP program, then the existing official band will be used and a new band does not need to be applied. b. Wing bands i. Wing bands are versatile, quick and easy to use. Wing bands can be used on larger sized Old English Bantam chickens up to adult turkeys and geese. If the band is locked in correctly, the band will stay in the wing for years and will remain clean and easy to read. Most small bantam owners will still want to use the small leg bands. ii. The wing band is loaded into the wing band pliers. The pointed end of the wing band tucks behind the bulge end of the pliers. The flat end of the wing band sits 1

2 on the circle imprint on the other end of the pliers. The pliers do not lock in the band like cow tag pliers, the pliers just hold it (barely). iii. The wing band is placed in the wing web of the bird. The largest surface area in the wing web is straight across from the elbow. Open up the wing, look on the underside of the wing, find the elbow, pluck the downy feathers in the wing web if necessary, and apply the band directly across from the elbow. I always look at the underside of the wing as I apply the band that way I don t hit any arm muscles (especially on the small sized birds) compared to blindly doing it from the top outer side of the wing. iv. Place the pliers/band so that the band number (on pointed side of the band) will be facing out, and that the locking end of the band is on the inside of the wing (less feathers to interfere with locking the band). Align the band along the wing web edge across from the elbow with the back end of the band just touching the wing web edge. Do not lock the band on too deep into the webbing that it is tight and pulling against the wing web edge (especially in growing birds) and do not apply it so loose that half of the band is hanging off the wing which may get caught up in the cage/pen wire. Once you are properly aligned, just close the pliers with a quick, firm motion like using a paper hole punch. Hand Shoulder Elbow 2

3 c. Regular leg bands i. Regular leg bands take a little longer to use, usually fall off within a year and get dirty and hard to read later on. SC NPIP leg bands come in small and regular sizes and require 2 separate banding pliers to lock the band (small and regular leg band pliers). ii. Before applying the leg band, bend the band into a circle. The more you mold the band into the size of the leg, the easier it is to lock with the pliers. iii. Place the band on the leg with the nipple underneath the holes. Align the nipple up through the correct hole. The flat end of the pliers goes under the band and the nipple end of the pliers is on top over the band. Place the plier s nipple into the nipple of the band and squeeze flat. If the band nipple is perfectly set within the band s hole, it will roll over and lock the band if not, the band comes off. iv. If there is an excess amount of the band, you can gently pinch one end to tighten the circle around the bird s leg. v. You may notice that the leg band pliers you purchase are too thick in order to easily fit the pliers between the bird s leg and band, especially with the small bird bands. You may need to grind down the outer edges of the pliers to make it thinner, as shown with the left pliers. 3

4 d. Tiny cinch or butt end leg bands i. These cinch bands are for smaller sized birds (quail, etc.) and should stay on the bird better than regular leg bands. SC cinch bands come in multiple sizes and each size requires its own separate banding pliers to lock the band. a. Size 6 small doves*, button quail? b. Size 8 bobwhite quail, doves* c. Size 10 homing pigeons* d. Size 12 OE bantam hens (not roosters), larger pigeons* *For traceability banding, pigeons and doves are not Pullorum tested. ii. Open up band before applying it. Place the closed band on the tip of the closed pliers and open up the pliers to crack open the band into a C shape, see picture A. iii. Place the band on the leg and close the band with the pliers as shown in picture B. (A) To open the cinch bands. (B) To close the cinch bands. 3. Perform the Pullorum Typhoid whole blood plate agglutination test. a. Place 1 drop of Pullorum Typhoid antigen on the test plate. Do not touch the end of the dropper to the plate or you will get more than 1 drop. Just let 1 drop fall from the dropper onto the plate. b. I like to use a needle and syringe rather than the poker that is on the loop tool to pierce the wing vein, since this edge becomes dull quickly. You can use the same needle for birds before it becomes dull and then you can change out the needle. I use 22 gauge needles for bantam sized birds and 20 gauge needles for anything larger. c. Use the needle to pierce the cutaneous ulnar vein as it comes across the elbow. The elbow bone serves a good hard surface to press against. Hold the needle/syringe perpendicular to the vein and punch straight down and up a drop of blood with come out. Adult male birds will have thicker skin over the vein making it harder to see and harder to pierce make sure you have a new sharp needle. Black skinned birds (silkies) 4

5 are also very hard to actually see the blue vein you can see the vein ridge as it crosses the elbow bone. You can also clip a toe nail and get fresh blood to do the test if you cannot get it from the elbow. Birds have very fragile veins and will develop a hematoma (blood under the skin) just warn the owner ahead of time it will heal up in about 2 weeks. Vein crossing elbow Vein crossing elbow d. Use the loop to collect the drop of blood and mix it in the antigen on the plate. Stirring is the most important part of this test! Stir the drop of blood into a quarter size circle about 12 times and then also pick up your plate and slowing tip the plate back and forth. e. Any agglutination or clumping seen within 2 minutes is a reactor. Any agglutination or clumping that forms after 2 minutes is not a reactor. If you are unsure if a reaction occurred within 2 minutes or longer, retest the bird and time it. The middle test is showing a (mild) positive agglutination that occurred within 2 minutes. 5

6 f. Rinse the needle and loop in the cup of water and pat dry on the paper towel before using on the next bird. Make sure there is no water remaining in the loop before you collect blood on the next bird or you will not get the correct amount of blood. 4. Record the information on the SC NPIP Pullorum Typhoid Testing Report a. Get a testing report form from Dr. Helm. b. Fill out the owner information and check the box for the reason for testing. c. Fill out the bird type, (NPIP bird code number only if this is for NPIP breeder Pullorum Clean certification), number of birds tested, number of pullorum reactors, and the band numbers. Separate the band numbers between different species of birds put chickens and their band numbers together, turkeys and their band numbers together, guineas and their band numbers together, etc. d. At the bottom, fill out the total number of birds tested, (breeder information only if testing is for NPIP breeder Pullorum Clean certification), sign your name as the tester, write your NPIP license number if you are a private licensed tester or your USDA accreditation number if you are a Type II veterinarian, and the date of testing. The flock owner only signs and dates the form if testing is for NPIP breeder Pullorum Clean certification and after agreeing to what is listed on the back side of the form. e. The top white copy is returned to Dr. Helm, the owner receives the 2 nd yellow copy, and the tester keeps the 3 rd pink copy for their records. 5. Pullorum plate reactors a. This plate test is a great screening test, but does not confirm that the bird has been exposed to Pullorum Typhoid. We do see false plate reactors. If you get a plate reactor, contact Dr. Helm. Blood needs to be collected from the reactor bird and the serum sent to the Clemson Diagnostic Veterinary Lab to perform another more specific test (Pullorum microagglutination test). There is no charge for this test for plate reactors. If you cannot collect the blood from the brachial vein along the humerus bone, then contact Dr. Helm to collect the sample. 6

7 Collecting blood from the brachial vein use a 3ml syringe and gauge needle. b. All Pullorum plate reactors and all the rest of the birds from that flock are restricted back at the owner s premises until further testing is completed. c. If the lab microagglutination test is also positive for antibodies, then the reactor bird must be submitted to the Clemson Diagnostic Veterinary Lab, euthanized and the internal organs cultured for Salmonella in order to confirm if the bird is positive for Pullorum Typhoid. d. Pullorum Typhoid Disease is a Reportable Disease to the State Veterinarian, so we have to complete the follow through testing in order to confirm it. 7

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