## 4 Conclusions  
The results of this study demonstrate that Tb-PAW can effectively reduce a range of bacteria involved in the development of DD. The level of reduction is depending on several parameters, including the exposure time and the treated bacterial species. Gram-positive bacteria (_S. aureus_ and _C. sporogenes_) were more resistant to the Tb-PAW than Gram-negative bacteria (_E. coli_, _F. necrohorum_ and _P. levii_). As the Tb-PAW in the present study could achieve a fast and high inactivation of the bacterial strains despite a neutral ph-value, the antimicrobial effect of the Tb-PAW seems to be only due to the action of reactive species \begin{table}
\begin{tabular}{c c c c} \hline \hline \multicolumn{4}{c}{**Storage (hours)**} \\ \hline
**Treatment** & **4** & **8** & **12** & **24** \\ \hline
7 \({}^{\circ}\)C treatment temperature & & & & \\ Control 1  & 4.93 \(\pm\) 0.81 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.02 \(\pm\) 0.74 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.71 \(\pm\) 0.59 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.05 \(\pm\) 0.63 \({}^{\mathrm{A}\,\mathrm{A}}\) \\ Tb-PAW 2  & 1.23 \(\pm\) 0.50 \({}^{\mathrm{B}\,\mathrm{B}}\) & 2.40 \(\pm\) 0.39 \({}^{\mathrm{B}\,\mathrm{B}}\) & 2.40 \(\pm\) 0.80 \({}^{\mathrm{B}\,\mathrm{B}}\) & 2.43 \(\pm\) 0.77 \({}^{\mathrm{B}\,\mathrm{x}}\) \\ \hline
21 \({}^{\circ}\)C treatment temperature & & & & \\ Control & 5.31 \(\pm\) 0.17 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.56 \(\pm\) 0.58 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.71 \(\pm\) 0.47 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.27 \(\pm\) 0.49 \({}^{\mathrm{A}\,\mathrm{A}}\) \\ Tb-PAW & 1.14 \(\pm\) 0.42 \({}^{\mathrm{B}\,\mathrm{B}}\) & 1.18 \(\pm\) 0.83 \({}^{\mathrm{B}\,\mathrm{B}}\) & 2.94 \(\pm\) 0.57 \({}^{\mathrm{B}\,\mathrm{B}}\) & 2.09 \(\pm\) 0.88 \({}^{\mathrm{B}\,\mathrm{x}}\) \\ \hline
30 \({}^{\circ}\)C treatment temperature & & & & \\ Control & 5.65 \(\pm\) 0.22 \({}^{\mathrm{A}\,\mathrm{A}}\) & 6.03 \(\pm\) 0.28 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.95 \(\pm\) 0.32 \({}^{\mathrm{A}\,\mathrm{A}}\) & 5.76 \(\pm\) 0.30 \({}^{\mathrm{A}\,\mathrm{A}}\) \\ Tb-PAW & 1.16 \(\pm\) 0.80 \({}^{\mathrm{a}\,\mathrm{B}}\) & 0.96 \(\pm\) 0.45 \({}^{\mathrm{a}\,\mathrm{B}}\) & 2.25 \(\pm\) 0.42 \({}^{\mathrm{a}\,\mathrm{B}}\) & 5.03 \(\pm\) 0.22 \({}^{\mathrm{b},\mathrm{B}}\) \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Means and standard deviation of _E. coli_ counts (log CFU/mL) following 1 min Tb-PAW treatment. within the Tb-PAW. Furthermore, Tb-PAW is maintaining high efficacy levels even under the presence of organic substances. Thus, our results indicate that Tb-PAW might constitute a potential DD-therapy in dairy cows. Due to the high time stability (12 h), even under elevated temperatures with up to 30 \({}^{\circ}\)C, Tb-PAW seems to be a promising tool being able to perform under real-life conditions in agricultural holdings, e.g., as a claw bath or spray solution. Further work in progress is focusing on the analysis of Tb-PAW applied directly to skin samples or in situ on wounds infected with DD. The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/app122312325/s1](https://www.mdpi.com/article/10.3390/app122312325/s1)]([https://www.mdpi.com/article/10.3390/app122312325/s1](https://www.mdpi.com/article/10.3390/app122312325/s1)), Table S1: Means and standard deviation of _Escherichia coli, Fusobacterium uncomporum and Porphomonas levii_ counts (log10 CFU/mL) after 1 min treatment of Tb-PAW, Tb-PAW with bovine serum albumin and Tb-PAW with sodium azide. Conceptualization, M.P., G.A., L.S., C.K., M.H. ([PERSON]) and V.G.-P.; methodology, V.G.-P., L.S. and G.A. ; software, V.G.-P.; validation, V.G.-P., L.S., M.P. and C.K. ; formal analysis, V.G.-P.; investigation, V.G.-P.; resources, C.O., M.H. ([PERSON]) and R.O. ; data curation, V.G.-P.; writing--original draft preparation, V.G.-P.; writing--review and editing, L.S., C.K., G.A., M.P., M.H. ([PERSON]), C.O., M.H. ([PERSON]), R.O., B.A., K.A.R., W.V. and L.t.B. ; visualization, V.G.-P.; supervision, M.P., L.S., C.K. and G.A.