Successfully reported this slideshow.
Your SlideShare is downloading. ×

Prevention and control of Mycoplasma sinoviae without vaccination

Prevention and control of Mycoplasma sinoviae without vaccination

Download to read offline

A presentation covering basic aspects regarding the prevention and control of Mycoplasma sinoviae (a poultry pathogen) without the use of vaccination.

Presented at the 2014 Biochek Seminar in Taiwan by Dr. Rafael Monleon

Contact me in LinkedIn for any question: www.linkedin.com/rafaelmonleon

A presentation covering basic aspects regarding the prevention and control of Mycoplasma sinoviae (a poultry pathogen) without the use of vaccination.

Presented at the 2014 Biochek Seminar in Taiwan by Dr. Rafael Monleon

Contact me in LinkedIn for any question: www.linkedin.com/rafaelmonleon

More Related Content

Related Books

Free with a 30 day trial from Scribd

See all

Related Audiobooks

Free with a 30 day trial from Scribd

See all

Prevention and control of Mycoplasma sinoviae without vaccination

  1. 1. Rafael Monleon, DVM, MSpVM, ACPV, PAS Business Unit Manager (Poultry) 16th January 2014 Taiwan Prevention and Control of MS Without Vaccination
  2. 2. Contents • Mycoplasma Synoviae • Diagnosis & Monitoring • Control – Cases
  3. 3. Mycoplasma Synoviae(MS) 1. Introduction 1) Most frequently occurs as a subclinical upper respiratory infection 2) Air sac lesions combined with ND, IB 3) Systemic infection with synovitis and CRD in chickens and turkeys 4) Not a virus and real bacteria, pleomorphic coccoid, approximately 0.2μm, fried egg type of colonial morphology
  4. 4. 2. History and Incidence 1) 1954 – Enlarged joint condition in poultry caused by an infectious agent(Olson et al) 2) 1964 – A respiratory form of MS infection occurs(Olson et al) 3) 1972 – Air sacculitis in broilers from MS combined with ND, IB(Kleven et al) 4) World wide distribution
  5. 5. 3. Characteristics 1) Generally weak pathogen of poultry 2) Strains vary in virulence 3) Economically damaging disease A. Layers: 5-10 eggs loss per year (MG: 10-20 eggs) B. Egg drop in lay C. Decreased hatchability and increased condemnations D. May cause mortality with respiratory disease, peritonitis and synovitis
  6. 6. 4) Chickens will remain infected after clinical signs have disappeared 5) Survival of MS (Room temperature) A. feather: 3 days B. nasal cavity: 12 hours C. other materials: less than 1 days 6) Eggshell Apex abnormality (EAA, since 2000) (South Africa, Japan, Holland, etc)
  7. 7. 4. Host 1) Chickens, turkeys, partridge, pheasants, peafowl, quail, guinea fowl, ducks and pigeons 5. Transmission 1) Vertical and horizontal infection 2) Faster spread than MG 3) Egg transmission rate appears to be highest during first 4-6 weeks after infection.
  8. 8. Diagnosis • Serology – Plate agglutination test – ELISA test – HI test • Molecular Tools • Isolation • Monitoring
  9. 9. 1) Plate agglutination test(or RSA) A. simple equipment B. fast procedure and detection of IgM (7DPI) C. become more difficult to get antigen D. Many false positives: after oil vaccination frozen serum DOC serum(dehydrated blood) just after eating up(gelatinized, lipoid change) hemolysis
  10. 10. 2) ELISA test A. Most common due to easy test and cheaper cost B. MS single or MG/MS combo kits is available C. Low ammount false positives sometimes D. Advantage to get massive test (3-5 plates per time) E. Mostly detect IgG / some IgM F. Biochek detects from 7 dpi and some previously undetectable variants
  11. 11. Mycoplasma Proficiency Results MS Field Isolate in 4 Wk Old SPF Chickens, 7 DPI* 2010 2011 RPA Ms** 33% 17% Mg/Ms(r) ELISA 100% 100% Other Mg/Ms ELISAs** 17% 44% % positives Assay *From the International PTS for Mg and Ms Report; GD B.V. Animal Health Services Ltd. , Netherlands ** various manufacturers Proficiency testing conducted by Animal Health Institute suggested early detection using this recombinant based Ms antigen in the Ms/Mg ELISA
  12. 12. Results (MS WVU1853 Inoculated) 0 20 40 60 80 100 0 7 9 14 16 %Positive Days Post-Challenge Mg/Ms(r) ELISA Mg/Ms (c) ELISA RPA (1&2) RPA (3)
  13. 13. Results (MS K5664 Inoculated) 0 20 40 60 80 100 0 7 9 14 16 %Positive Days Post-Challenge Mg/Ms(r) ELISA Mg/Ms (c) ELISA RPA (all 3)
  14. 14. 3) HI test A. used to be the golden standard to confirm positive of RSA or ELISA B. HI test set up is rare Need live antigen (mycoplasma) C. more labs are using PCR for confirmation of serological test
  15. 15. 2. PCR 1) Become more common in many countries 2) Not perfect, but more accurate than serological test 3) Be careful for cross contamination 4) Re-check with serological test after 3-4 weeks for re- confirmation 5) Tracheal, orbital, choanal swabbing 6) Keep samples into PBS and deliver to lab on the sampling day with ice
  16. 16. 7) Sequencing for epidemiology A. If there is frequent infection, sequencing will be very helpful e.g. K1858(US origin), EsPk1UAF08(Pakistan), B45/04(UK), 94011(US), VlhA2.28.1, WVU1853, etc.
  17. 17. 3. Isolation 1) Difficult and skillful procedure to perform 2) Only few Mycoplasma researchers have the set up 3) Most of confirmation is finished at PCR with serological background
  18. 18. Monitoring Program 1) Monitor every 3-4 weeks life of flock 2) Seroconversion in general is not well developed before 6-7 weeks old However if infections occurred before 6-7 weeks, further monitoring is required 3) 3 weeks interval will be matched to incubation period of embryo, so preventing distribution of positive DOC to customers 4) Example (weeks old): 1,7,10,14,18,22,26,30 (4 weeks interval),33,36,39,42,45,48,51,54,57,60,63 (3 weeks interval), 19 times
  19. 19. 5) At least 18-23 sera from each house, if take sera from only one house, you may miss first infection 6) Always thinking about Cost-Effective
  20. 20. Case of infection 1. Case 1: faster seroconversion 1) ELISA result (BioChek, Cutoff = 668) 39weeks old 42 weeks old House 1 2 3 4 1 2 3 4 1 207 1 50 128 330 11,742 55 64 2 63 4 260 82 414 8,407 99 153 3 75 252 182 439 38 8,903 261 7 4 44 29 85 168 178 8,127 394 38 5 332 214 118 32 69 13,931 245 239 6 69 101 75 47 47 12,608 93 84 7 172 10 172 85 108 5,464 354 93 8 394 66 60 145 72 4,757 297 141 9 420 368 853 182 1 3,056 105 87 10 391 53 168 32 5 14,314 344 527 11 175 1,312 41 168 20 13,782 404 120 12 38 158 186 214 47 6,626 1 1 13 688 18 1 175 330 13,875 102 226 14 23 47 23 72 108 2,349 15 129 15 214 179 114 20 6,674 384 Positive 1/14 1/15 1/15 0/15 0/15 15/15 0/14 0/15 PCR Negative PCR Positive Phase 1: 12-21 days(1st Ab) Phase 2: 1-21days(5-10%) Phase 3: 7-32days(90-95%) Phase 4: 3-19days(100%) From 23 to 93days
  21. 21. 2) Presumption of infection period Phase 1: 12-21 days(1st Ab) Phase 2: 1-21days(5-10%) Phase 3: 7-32days(90-95%) Phase 4: 3-19days(100%) From 23 to 93days From 11 to 72days after 1st Ab Infection Aug,28 Sep,4 Sep,11 Sep,18 Sep,25 Oct,4 Oct,11 1st Ab detection 100% Positive Less than 21days Between Aug,21 and Sep,1 ? PCR +ve, Oct,8 EsPk1UAF08
  22. 22. 3) Epidemiology A. PCR –ve when 1st antibody detected B. Investigate visitor book and disinfection record C. Contact with poultry people D. Wild bird E. Repair procedure F. Sequencing: EsPk1UAF08 strain < Visitor book >
  23. 23. 4) Route of infection A. Failure to find source of infection in many case B. assuming very high dust in house #2 may facilitate MS infection 5) Treatment A. Enrofloxacin application to postpone spread to other houses B. Change order of visit from farm to farm
  24. 24. 2. Case 2: Slower seroconversion 1) ELISA result 22 weeks old 26 weeks old 30 weeks old 33 weeks old House 1 2 3 1 2 3 1 2 3 1 2 3 1 94 10 10 6 2,713 10 5,947 784 551 481 2,315 373 2 10 10 10 10 2,063 10 4,056 4,003 336 507 755 969 3 53 10 74 12 930 10 633 1,618 518 4,331 809 341 4 75 10 10 167 407 10 374 944 138 1,503 350 672 5 10 741 10 9 1,604 10 273 755 41 1,731 7,061 429 6 10 203 10 38 1,136 10 5,185 485 1,126 4,243 6,700 318 7 12 259 10 10 6 10 981 1,049 711 274 2,012 944 8 10 10 10 4 569 86 148 792 198 1,149 2,012 1,019 9 71 10 10 610 497 10 303 4,356 268 1,597 5,559 304 10 113 10 10 10 10 10 1,101 1,980 10 2,336 5,914 1,166 11 10 45 10 10 2,890 10 205 956 2,517 993 5,789 1,435 12 10 29 10 10 6,382 10 237 842 804 186 6,193 1,197 13 10 10 10 10 1,107 3,153 84 534 1,513 1,167 12,842 745 14 10 10 59 10 4,758 99 2,263 1,087 1,019 1,002 2,418 354 15 10 10 10 48 493 40 1,265 142 1,990 1,807 2,864 Positive 0/15 1/15 0/15 0/15 8/15 1/15 7/15 12/15 7/15 11/15 13/14 9/15 % 0% 6.7% 0% 0% 53.3% 6.7% 46.7% 80.0% 46.7% 73.3% 92.8% 60.0%
  25. 25. 2) Presumption of infection period Phase 1: 12-21 days(1st Ab) Phase 2: 1-21days(5-10%) Phase 3: 7-32days(90-95%) Phase 4: 3-19days(100%) From 23 to 93days From 11 to 72days after 1st Ab Infection Nov,7 Nov,17 Dec,8 Jan,5 Feb,2 Feb,23 Mar,24 1st Ab detection 22wks old 92.8% Positive 33wks old 77days (11weeks) Between Nov,7 and 17 ? PCR +ve, B45/04
  26. 26. 3) Epidemiology A. No relationship with previous strain (new strain) B. 12 visits from out of farm C. Less visitors and disinfection than laying farm 4) Route of infection A. Failure to find a source of infection 5) Treatment A. Enrofloxacin application to postpone spread to other houses B. Change order of visit from farm to farm
  27. 27. Control • Biosecurity • Treatment – Cases • Vaccination
  28. 28. 1.- Biosecurity 1) Visit Authorization 2) Shower-in, shower-out 3) Fumigation 4) Vehicle disinfection unit 5) Feed transporting system 6) Wild bird control
  29. 29. Vehicle disinfection Hand sanitation Authorization 1) Authorization
  30. 30. Change shoes and hand disinfection Spraying disinfectant mist and boots disinfection Take off private clothes Shower Put on working clothes Inside of farm 2) Shower-in, shower-out
  31. 31. All things from outside to inside of farm, must be disinfected with peracetic acid through procedure of spray of 20um particle-> disinfection -> exhausting for 30 minutes 3) Fumigation
  32. 32. 4) Vehicle disinfection unit
  33. 33. 5) Feed transporting system
  34. 34. 6) Wild bird control A. Remove all trees inside of farm B. Remove all vegetation inside of farm C. Remove feed spilt
  35. 35. 2.- Treatment 1) Generally antibiotics treatment is totally successful 2) But, we can find several success story with antibiotics A. Cost a lot B. Cage system in laying period C. Floor system in rearing period D. Floor system in laying period (Fiorentin et al, 2003)
  36. 36. Eradication in cage laying system A. History - open-sided house, 4 lines of A type cage - 6 houses can hold 10,000 birds respectively - multi-age, artificial insemination
  37. 37. B. Multi-aged flocks - New flock is replaced every 4~6 month at the age of 12 to 14 weeks old. - Jun 2002 flock* => Dec 2002 => Mar 2003 => July 2003** => Mar 2004 => Jun 2004 => Oct 2004 *Red colored flocks are MS positive ** Black colored flocks are MS negative < House arrangement and flock held situation> 4th: Dec 2002 flock => Mar 2004 flock 3rd: Dec 2002 flock => Mar 2004 flock 2nd: Mar 2003 flock => Jun 2004 flock 1st: Mar 2003 flock => Jun 2004 flock
  38. 38. C. Treatment - Feed additives : 10mg/kg of Chlortetracycline - Drinking water : 10mg/kg Doxycycline 10mg/kg Enrofloxacin 7.5mg/kg Tilmicosin - Tilmicosin : every 6 weeks administration program - Others: between Tilmicosins
  39. 39. D. Results Week Flock 2003 2004 15 20 25 26 29 34 38 42 47 51 2 5 7 14 18 20 22 24 26 28 31 Jun, 2002 + - - - Dec, 2002 + - + + + - + + + - - + - Mar, 2003 - - - + + + - - - - - - - Jul, 2003 + + - - - - - - - - - - - - Mar, 2004 - - - - Mean ELISA titer of each flock 1 10 100 1000 10000 Jun-03 Jul-03 Aug-03 Sep-03 O ct-03 Nov-03 Dec-03 Jan-04 Feb-04 M ar-04 Apr-04 M ay-04 Jun-04 Jul-04 Date Titer Jun, 2002 Dec, 2002 Mar, 2003 July, 2003 Mar, 2004 Positive cutoff 594
  40. 40. E. Summary 2003 Antibiotics (for 3days, mg/kgB.W) Flock DateJun,2002 Dec,2002 Mar,2003 July,2003 Mar,2004 age PCR ELISA age PCR ELISA age PCR ELISA age PCR ELISA age PCR ELISA 15wks Til. 7.5 43 + 18 + 4/14 20wks Til. 7.5 48 - 22 - 5/19 22wks CTC 10 50 9/10 24 10/10 6/2 25wks CTC 10(14d) 53 - 27 + 13 - 6/23 26wks Enro. 10(5d) 54 - 10/10 28 + 10/10 14 - 0/10 6/30 Doxy. 10(5d) 29wks 31 + 9/10 17 0/10 7/21 34wks Depleted 36 - 10/10 22 - 0/10 8/25 36wks Doxy. 10(5d) 9/8 38wks Enro. 7(5d) 40 + 10/10 26 + 1/10 9/22 42wks Til. 7.5 44 + 10/10 30 + 3/10 15 + 10/20 47wks Til. 7.5 49 + 10/10 35 + 7/10 20 + 0/10 11/24 51wks 53 - 10/10 39 - 7/10 24 - 0/10 12/25 2004 1wks Til. 7.5 56 42 26 1/7 2wks Doxy. 10 57 - 10/10 43 - 9/10 27 - 0/10 1/14 5wks Enro. 10 60 + 46 - 30 - 2/4 7wks Til. 7.5 62 - 10/10 48 - 10/10 32 - 0/10 2/18 9wks 64 50 34 3/2 13wks Til. 7.5 Depleted 54 10/10 38 0/10 3/31 14wks Doxy. 10 55 - 10/10 39 - 0/10 4/7 18wks Enro. 10 59 - 43 - 5/5 19wks Til. 7.5 60 44 5/12 20wks 61 - 8/10 45 - 0/10 5/19 22wks Depleted 47 - 6/2 24wks 49 - 0/10 14 - 0/10 6/16 26wks 51 - 16 - 6/30 28wks 53 - 18 - 7/14 31wks 56 - 0/10 21 - 0/10 8/5
  41. 41. A. History - Transovarian infection is suspected (GP flock is seroconverting) - 3 houses of 30,000 birds - closed house Eradication in floor system Case #1
  42. 42. B. Treatment - Drinking water: 3 days/week, 5.0mg/kg BW - From 1 week to 20 weeks old
  43. 43. C. Result - Mother flock’s ELISA: 34.1% Positive Seroconversion is in progress Age(W) House 1 2 3 PCR 1 5/15(33%) -ve 5 0/15 0/15 0/15 -ve 7 0/15 0/15 1/15 10 0/15 0/15 0/15 -ve 14 0/15 0/15 0/15 -ve 18 0/15 0/15 0/15 -ve 22 0/15 0/15 0/15 -ve
  44. 44. D. Comments - 2 flocks successful in a row - 3rd flock was failed(isolate same strain with mother flock) - 0.3USD/bird
  45. 45. Eradication on floor rearing Case #2 • Day 0 – MS positive • Tylosin @ 3d for 5 days • Tylosin @ 3 wks every 4 weeks • Results seems to have eliminated serological evidence by 3 & 10 weeks
  46. 46. Eradication on floor rearing #2 DOC
  47. 47. Eradication on floor rearing #2 3wks National  Chiayi  University Report:    Histogram/Blockdiagram Dr.  Kuo  Lab ResultSample  ID Well Raw  OD S/P  Ratio Titer Titer  Group Reason  : Housenumber  : smallCode  : Type  : 21  Day(s)Age  : 2013/10/30Samplingdate  : Company  : G.P.Customer-­Name  : 2013/10/30    07:24:37Lab  code  : Assay: Bleedingdate: Lotnumber: Testdate: 0 2013/10/30 Ms 2013/10/30 10/0/0Neg/Sus/Pos: 10Mean  Titer: Min-­Max  Titer: GMT: %CV: 50 1  -­ 138 23 88 No.  Samples: Target  Titer: Target  %CV: VI  Index: 0 Target  Range  VI: Interpretation  VI: RF10  (2910)Meantiter  Ref.Controls RF10  (1500-­4500)Titer  Range  Ref.Controls Positive  Cuttoff  S/P: 0.5>= 0.584D01+ 0.608C01+ 0.125B01-­ 0.104A01-­ 01 F01 0.191 0.159 138 0 neg 02 G01 0.157 0.088 65 0 neg 03 H01 0.164 0.103 79 0 neg
  48. 48. Eradication on floor rearing #2 10wks
  49. 49. Serological Profile Following Treatment
  50. 50. Serological Profile Following Treatment
  51. 51. Serological Profile Following Treatment
  52. 52. Control 3.- Vaccination 1) Killed and live vaccine are licensed A. Killed vaccine can’t protect infection 2) MSH Live vaccine is from Bioproperties now registered in more and more countries A. No vertical transmission evidence B. Not pathogenic for chickens C. Chemical mutagenesis treated
  53. 53. Contents • Biochek Software – Field Interface – Multiuser-Cloud Database • VDP • Collaborative Research Projects
  54. 54. Acknowledgements • Dr. Young Ho Hong • Ms. Wendy Wu
  55. 55. THANK YOU rafaelmonleon@biochek.com

Editor's Notes

  • 8
  • 27
  • 53
  • 54
  • 55

×