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Analysis of Lamb Vaccination Methods

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Serum group

Number of individuals

Number of serum samples investigated

Characteristics

Additional notes

Vaccinated lambs

4

4

Lambs from the vaccine safety trial received prototype C. pecorum vaccine at the time of birth and 12 weeks post-birth

Serum samples were tested at 24 weeks post-birth

Naturally infected lambs (Clearance of  Infection at ten months)

13

39

Lambs from a commercial farm in NSW, Australia with endemic C. pecorum infections, vaginal or rectal or ocular swabs positive for C. pecorum by qPCR and serology positive by CFT

Samples were tested for three different time points of the study period i.e. at two, six and 10 months of age

Naturally infected lambs (Recurrent infections)

5

15

Lambs from a commercial farm in NSW, Australia with endemic C. pecorum infections, vaginal or rectal or ocular swabs positive for C. pecorum by qPCR and serology positive by CFT

Samples were tested for three different time points of the study period i.e. at two, six and 10 months of age

Naturally infected sheep with polyarthritis

19

19

Sheep and lambs across Australian flocks in NSW with clinical evidence of polyarthritis and C. pecorum serology positive by CFT

Cross-sectional sampling

Negative and specificity control groups

4

4

Uninfected lambs from a commercial farm in NSW, Australia with endemic C. pecorum infections, vaginal, rectal and ocular swab negative for C. pecorum by qPCR and serology negative by CFT

Cross-sectional sampling

Negative and specificity control groups

4

4

Pre-immunized lambs from the vaccine safety trial received  prototype C. pecorum vaccine

Samples were tested at the time of birth when lambs received their first vaccine dose

Total

41

77

Table 1. Description of origin, C. pecorum status and sampling characteristics of serum panel used in the study.

Table 2. Characteristics of C. pecorum detection by PmpG IgG ELISA (EPT), qPCR (copy no.) and CFT (serotiter), percentage positivity, percentage distribution of lambs exhibiting low, intermediate and high titer/copy no. detected by PmpG IgG ELISA, qPCR and CFT[SB1].

Groups

PmpG IgG positive

qPCR positive

CFT positive

Cleared infections

69.23% (27/39)

33.33% (13/39)

46.15% (18/39)

Recurrent infections

93.33% (14/15)

60% (9/15)

53.33% (8/15)

Polyarthritis

89.47 (17/19)

NA

100% (19/19)

Total

79.45% (58/73)

40.74% (22/54)

61.6% (45/73)

Titer/copy no.

PmpG IgG EPT

qPCR Load/copy no.

CFT serotiter

Low

20.54% (15/73)

22.22% (12/54)

8.88% (4/45)

Intermediate

43.83% (32/73)

7.40% (4/54)

66.66% (30/45)

High

15.06% (11/73)

11.11% (6/54)

24.44% (11/45)

Anti-PmpG IgG EPT cut-off >500, low <2000, intermediate <10,000, high ≥10,000; PCR Load/copies cut-off >10, low <100, intermediate <1000, high ≥1000; CFT serotiter cut off >8, low <32, intermediate <128, high ≥128.

Table 3. Epitopes recognised with 100% frequency, <100% ,≥70% frequency by vaccinated lambs, naturally infected lambs, naturally infected lambs that are high responders of anti-PmpG IgG, low responders of anti-PmpG IgG and naturally infected lambs with polyarthritis.

Serum group

N

Epitopes with 100% frequency

Epitopes with <100% ,≥70% frequency

Mean ± SD absorbance of epitopes (100% FD)

Mean ± SD absorbance of epitopes (<100% ,≥70% FD)

Mean ± SD absorbance of test/control peptides

Vaccinated lambs

4

72

11,15,16,17,79

P72 0.966±0.366

P11 0.791±0.307, P15 0.982±0.499, P16 1.573±0.807, P17 1.111±0.432, P72 0.966±0.366, P79 1.046±0.559

0.157±0.023

Naturally infected lambs

8

None

9,10,16,17,21,72

None

P9 0.575±0.284, P10 0.537±0.551, P16 0.854±0.461, P17 0.552±0.349, P21 0.955±0.47, P72 1.923±0.321

0.157±0.023

Naturally infected lambs;

High responders of anti-PmpG IgG

4

21

10,16,17,22,23,72

P21 0.861±0.656

P10 1.081±0.835, P16 0.990±0.694, P17 0.878±0.521, P22 0.755±0.486, P23 0.829±0.708, P72 0.604±0.470

0.157±0.023

Naturally infected lambs;

Low responders of anti-PmpG IgG

4

9,16,17,72,78

10,21,33,34,35,77,79

P9 0.429±0.248, P16 0.294±0.086,       P17 0.350±0.107, P72 0.275±0.079,     P78 0.218±0.046

P10 0.283±0.139, P21 0.257±0.040, P33 0.289±0.106, P34 0.290±0.047, P35 0.270±0.073, P77 0.237±0.068, P79 0.22±0.058

0.136±0.053

Naturally infected sheep with polyarthritis

4

17,21

9,16,22,23,24,32,33,72,79

P17 0.473±0.128, P21 0.415±0.162

P9 0.613±0.189, P16 0.489±0.137, P22 0.335±0.088, P23 0.364±-.072, P24 0.393±0.123, P32 0.394±0.073, P33 0.428±0.089, P72 0.36±0.044, P79 0.435±0.217

0.188±0.095

Epitopes common across all groups

12

None

72 (91.7%), 16, 17 (83.3%)

None

P72 0.552±0.413, P16 0.969±0.689, P17 0.661±0.431


[SB1]Analyses to be carried out on sensitivity, specificity and lag. Can be incorporated in this table.


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