1. United States Department of Agriculture Animal and Plant Health Inspection Service Surveillance for Bovine Tuberculosis in U.S. Livestock Tuberculosis Eradication Program Ruminant Health Programs Updated April 28, 2009

2. United States Department of Agriculture Animal and Plant Health Inspection Service Overview  Affected herds  Slaughter surveillance  Slaughter inspection  Granuloma lesions submitted for histopathology  Mexico origin cases  Live animal surveillance  Tuberculin skin testing  Gamma interferon testing  M. bovis in swine, Molokai Island, Hawaii

3. United States Department of Agriculture Animal and Plant Health Inspection Service Affected Herds* FY 2000–2009, United States *80 herds - 64% beef, 28% dairy, 2% mixed, 5% captive cervid

4. United States Department of Agriculture Animal and Plant Health Inspection Service Affected Herds By State, FY 2000–2009 Affected Herds *81 herds - 64% beef, 28% dairy, 2% mixed, 5% captive cervid

5. United States Department of Agriculture Animal and Plant Health Inspection Service Live Animal vs. Slaughter Surveillance — Finding Affected Cattle Herds, U.S., FY 2000 – 2009 Michigan, Minnesota (54) Rest of U.S. (28) Total affected cattle herds: 75 *No herd: 7 adult slaughter cases from California, Texas, Nebraska, New Mexico, South Dakota — no affected herd was detected

6. United States Department of Agriculture Animal and Plant Health Inspection Service Granuloma Submissions from U.S. Slaughter Plants, 2000–2008 Submissions Fiscal Year

7. United States Department of Agriculture Animal and Plant Health Inspection Service Granuloma Submission Rates for Slaughtered Adult Cattle, 2000–2008 Submissions continue to exceed target! Target: 5/10,000 adult cattle slaughtered Target Rate Fiscal Year

8. United States Department of Agriculture Animal and Plant Health Inspection Service Granuloma Lesion Submission in “Top 40” Adult Cattle Slaughter Plants, FY 2008 VI AKHI PR < 5 lesions/10,000 killed (7) - 133% increase from FY07  5 lesions/10,000 killed (33) Modified accredited advanced status Modified accredited status

9. United States Department of Agriculture Animal and Plant Health Inspection Service M. bovis Positive Cattle From Slaughter Tissue Sample Submissions, 2001–2009 358 TB cases in 9 years!! Updated 4/28/2009

10. United States Department of Agriculture Animal and Plant Health Inspection Service  18 positive by histology  17 cattle cases  11 confirmed M. bovis  PCR or culture  5 – other Mycobacterium  M. avium (4), M. kansasii (1)  1 – PCR negative, no culture  1 confirmed M. bovis – captive elk Confirmed Cases of M. bovis from Slaughter Surveillance, FY 2009 Updated 4/28/2009

11. United States Department of Agriculture Animal and Plant Health Inspection Service  6 adult cases  3 from 1 MN buyout herd  North Dakota – infection not confirmed in herd  PA slaughter case – no ID, multiple traces/states under investigated  NE adult beef cow under investigation (new)  1 captive elk, NE – herd confirmed infected  5 cases in fed cattle  2/5 (40%) Mexican origin  1 – Chihuahua  1 – Veracruz  1 – roping steer SE US  2 – under investigation Confirmed Cases of M. bovis from Slaughter Surveillance, FY 2009 Updated 4/28/2009

12. United States Department of Agriculture Animal and Plant Health Inspection Service  42 cattle positive by histology  Of these,  34 – confirmed M. bovis (PCR, culture)  6 – other Mycobacterium  M. avium [5], M. intracellulare [1]  2 – negative Confirmed Cases of M. bovis from Slaughter Surveillance, FY 2008

13. United States Department of Agriculture Animal and Plant Health Inspection Service  1 adult case, CA  33 cases in fed cattle  9/33 (27%) Mexican origin  4 – Chihuahua, 2 – Coahuila, 2 – Tamaulipas  1 – under investigation  2 from Canada  19 – Feedlot, TX  1 – Domestic case, NE  MN  3 – Investigation pending Confirmed Cases of M. bovis from Slaughter Surveillance, FY 2008

14. United States Department of Agriculture Animal and Plant Health Inspection Service 33 fed 1 adult 19 TX feedlot cases—assembled from KS/OK sales Location of 34 Cattle Positive for M. bovis Detected at Slaughter October 1, 2007–September 30, 2008

15. United States Department of Agriculture Animal and Plant Health Inspection Service Outbreak of TB in TX Feedlot FY 2008  19 TB cases in two feeder groups  Origin of cattle were old rodeo steers (many “M” branded) assembled from sales in OK and KS  Infected steers exposed 3,000 heifers backgrounding in same feedlot (shared hospital pen)  Epi tracing completed for 300– 600 heifers potentially exposed

16. United States Department of Agriculture Animal and Plant Health Inspection Service Genotyping in Epidemiologic Investigations Analysis of differences in nucleic acid sequences between strains of same species  Spoligotyping  Fast, but lacks discriminatory ability  RFLP–IS6110, PGRS (two types)  High discrimination, technically demanding, pattern comparison difficult when many strains  Mycobacteria Laboratory, NVSL is routinely genotyping M. bovis strains to assist investigations

17. United States Department of Agriculture Animal and Plant Health Inspection Service  California, three affected herds  Strain isolated from Herd 2 different from Herds 1 and 3  CA, TX slaughter cases, 2 cases from Campeche  Herds 1 and 3, strain identical  Most similar to CA slaughter case FY97 and NM affected herd FY 08  Feedlot outbreak with 19 affected cattle  All isolates identical — point source outbreak  Retired roping/rodeo cattle  Strains occurs infrequently in database — most similar to MX origin slaughter cases and untraceable SD case FY 05  New York deer  Close match to early 1990’s NY cervid strains  Comparison of MN strains, 2005–2008  From deer and cattle  All strains identical or minor variations Genotyping Results, FY 2008

18. United States Department of Agriculture Animal and Plant Health Inspection Service TB Cases Identified per 100,000 Cattle Imported from Mexico, 1995 – 2008

19. United States Department of Agriculture Animal and Plant Health Inspection Service Cattle Imports from Mexico By Import Cycle Year* 2003–2007 *Import cycle year ending August 31 Data source: Mexican Cattlemen’s Association

20. United States Department of Agriculture Animal and Plant Health Inspection Service 2 2 Origin of Eight TB Cases with Mexican Eartags, FY 2008 4 Reviewed!

21. United States Department of Agriculture Animal and Plant Health Inspection Service Time Spent in U.S. Cattle of Mexican Origin

22. United States Department of Agriculture Animal and Plant Health Inspection Service Caudal Fold Test Performance Cattle and Bison, FY 2006–2008 *As of October 15, 2008, 48 States FY2006FY2007FY2008* Total tests1,106,6691,135,7251,366,186 Responders10,91616,33620,229 Percentage, overall1.0 %1.4 %1.5% Percentage, by state (> 300 tests) 0.0 – 3.3 %0.0 – 3.2 %0.0 – 3.9 % States > 1%111512 States < 0.25%111213

23. United States Department of Agriculture Animal and Plant Health Inspection Service Caudal Fold Test Results in Cattle By State,* FY 2008 State *States with > 300 tests (n=1,366,004 tests, 47 States and Puerto Rico) Source: AWBDS and States < 0.05% in 6 States; 25,000 cattle

24. United States Department of Agriculture Animal and Plant Health Inspection Service Bovine Gamma Interferon Testing FY 2006–2008 FY 2008  15,601 tests  Includes repeat testing of individuals  94% of tests for cattle from 5 States  CA, MI, MN, NM, TX  Cattle tested from 20 states  Approved testing labs unchanged: TX, MI, CA, NVSL Annual tests by fiscal year, 2006-2008

25. United States Department of Agriculture Animal and Plant Health Inspection Service  Increase in positives beyond expected  Spring 2008, two labs, dairy cattle—CA, NM  Avian tuberculin—new lot with decreased ability to stimulate cells during assay  Above does not fully explain findings  Additional hypotheses include “real” immune phenomena in cattle, conditions after sample drawn  Invalid results – cells do not stimulate  Rare, but requires repeated gamma testing or CCT to resolve  Manufacturer (Prionics) has identified need to find new source for stimulating bovine and avian antigens  Once antigen source identified, plan to evaluate new antigens side- by-side with currently used antigens  Currently, antigens sourced from Australia (CSL) Bovine Interferon Gamma Testing FY 2008

26. United States Department of Agriculture Animal and Plant Health Inspection Service Bovine Interferon Gamma Test Results By Testing Laboratory, FY 2008 LaboratoryNumber PositiveNumber TestedPercent Positive California1655,1623.2% Michigan1532,7915.5% Texas1726,7622.5% NVSL268862.9% Total51615,6013.3%

27. United States Department of Agriculture Animal and Plant Health Inspection Service Tuberculin Testing of Cervids FY 2006–2008 FY 2006FY 2007FY 2008 Total cervids tested 25,42110,35319,147 SCT Suspects 374231287 Response rate 1.5%2.2%1.5% Source: AWBDS; as of October 8, 2008 for FY 2008

28. United States Department of Agriculture Animal and Plant Health Inspection Service Surveillance for M. bovis in Feral Swine, Molokai Island, Hawaii, FY 2007–2008  2008: tissues submitted for 59 animals; 7 confirmed*  2007: 6 confirmed*  Feral swine reservoir  Cattle continue to test negative *Confirmed by PCR or culture

29. United States Department of Agriculture Animal and Plant Health Inspection Service Lymph node, culture positive for M. bovis Thank You Questions or Comments?