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
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. 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. 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. 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. 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. Diagnosis
• Serology
– Plate agglutination test
– ELISA test
– HI test
• Molecular Tools
• Isolation
• Monitoring
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. 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. 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
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. 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. 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. 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. 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. 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
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. 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. 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
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. 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
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. 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
34. 6) Wild bird control
A. Remove all trees inside of farm
B. Remove all vegetation inside of farm
C. Remove feed spilt
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. 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. 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. 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
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
44. D. Comments
- 2 flocks successful in a row
- 3rd flock was failed(isolate same strain with mother
flock)
- 0.3USD/bird
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
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