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. 2025 Feb 16;15(4):570.
doi: 10.3390/ani15040570.

Evaluating the Effectiveness of Probiotic and Multivalent Vaccination Strategies in Mitigating Bacterial Chondronecrosis with Osteomyelitis Lameness Using a Hybrid Challenge Model

Amanda Anthney  1 , Khawla Alharbi  1   2 , Ruvindu Perera  1   2 , Anh Dang Trieu Do  1   2 , Andi Asnayanti  1 , Reginald Onyema  3 , Sara Reichelt  3 , Antoine Meuter  4 , Palmy R R Jesudhasan  1   2   5 , Adnan A K Alrubaye  1   2
Affiliations

Evaluating the Effectiveness of Probiotic and Multivalent Vaccination Strategies in Mitigating Bacterial Chondronecrosis with Osteomyelitis Lameness Using a Hybrid Challenge Model

Amanda Anthney et al. Animals (Basel). .

Abstract

Bacterial chondronecrosis with osteomyelitis (BCO) is caused by several bacteria, including Salmonella, Staphylococcus spp., Escherichia coli, Enterococcus spp., and Mycoplasma spp., and BCO is a significant animal health and welfare issue in broiler production, causing 1-2% of bird condemnation at marketing age and resulting in annual losses of tens of millions of dollars. This study evaluated the efficacy of a probiotic program alone and combined with a multivalent electron beam (eBeam)-inactivated vaccine in reducing BCO lameness. The probiotic program included an Enterococcus faecium spray (E. faecium 669, at 2 × 109 CFU/bird) at hatch and a triple-strain Bacillus-based product (B. subtilis 597, B. subtilis 600, and B. amyloliquefaciens 516 at 1 × 109 CFU/bird/day) in drinking water from day 1 to day 56. An aerosol transmission challenge model simulated commercial bacterial exposure. Birds were divided into five groups: a positive control (PC) group (T1) and a negative control (NC) group (T2) receiving no treatment and three treatment groups receiving the probiotic program (T3), the multivalent vaccine (T4), or both the probiotic program and the multivalent vaccine (T5). Data analyzed via ANOVA (p < 0.05) showed T3, T4, and T5 had significantly lower lameness (43.7%, 40.3%, and 40.7%) than T2 (71.0%) and T1 (83.0%). T5 resulted in reductions comparable to T4, indicating no significant synergistic effect. These findings show that probiotics alone or with a vaccine effectively mitigate BCO lameness, enhance broiler welfare, and reduce economic losses.

Keywords: BCO; broilers; eBeam-inactivated vaccine; lameness; probiotic.

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Conflict of interest statement

Reginald Onyeme and Sara Reichelt are employees of Aviagen North America. Antoine Meuter is an employee of Novonesis Group. The authors declare that this study received funding from Aviagen North America and Novonesis Group. Aviagen North America provided fertile eggs, and Novonesis Group supplied feed additives. The funders were not involved in the study design, data collection, analysis, interpretation, manuscript writing, or the decision to publish this manuscript. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure A1
Figure A1
The experimental treatment layout diagram Treatments are marked as follows: PC = positive control, T2 = negative control, T3 = GALLIPRO® Hatch/Fit, T4 = multivalent eBeam vaccine, and T5 = GALLIPRO® Hatch/Fit and multivalent eBeam vaccine.
Figure 1
Figure 1
Classifications of femoral head lesions for BCO lesion development. The arrows indicate the affected regions in each stage of lesion progression: (A) N (normal); (B) FHS (femoral head separation); (C) FHT (femoral head transitional degeneration); (D) FHN (femoral head necrosis).
Figure 2
Figure 2
Classifications of tibial head lesions for BCO lesion developments. The arrows indicate the affected regions in each stage of lesion progression: (A) N (normal); (B) THN (tibial head necrosis); (C) THNS (tibial head necrosis severe); (D) THNC (tibial head necrosis caseous); (E) TD (tibial dyschondroplasia).
Figure 3
Figure 3
Percentage cumulative lameness by treatment groups from d36–57 of this study. (a–c) Values within a category with different letters vary significantly at p < 0.05 on day 56 (GLM analysis). Treatments are marked as follows: NC = negative control, PC = positive control, T3 = GALLIPRO® Hatch/Fit, T4 = multivalent eBeam vaccine, and T5 = GALLIPRO® Hatch/Fit and multivalent eBeam vaccine.
Figure 4
Figure 4
Distribution of femoral and tibial head lesion severity across treatment groups. Lesion categories, based on Wideman (2016) [2], include normal femoral and tibial head (N), femoral head separation (FHS), femoral head transitional lesions (FHTs), femoral head necrosis (FHN), tibial head necrosis (THN), tibial head necrosis severe (THNs), tibial head necrosis caseous (THNC), and tibial dyschondroplasia (TD). Treatment groups are labeled as follows: T1 = positive control, T2 = negative control, T3 = GALLIPRO® Hatch/Fit, T4 = multivalent eBeam vaccine, and T5 = GALLIPRO® Hatch/Fit combined with multivalent eBeam vaccine.
Figure 5
Figure 5
Weekly cumulative mortality across treatment groups. Treatment groups are labeled as follows: NC = negative control, PC = positive control, T3 = GALLIPRO® Hatch/Fit, T4 = multivalent eBeam vaccine, and T5 = GALLIPRO® Hatch/Fit combined with multivalent eBeam vaccine.
Figure 6
Figure 6
Distribution of bacterial identifications across different tissue types of lame birds in all treatments.
Figure 7
Figure 7
Total isolate count (%) of bacterial species isolated from blood, femur, and tibial of lame birds.
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