## 2 Materials and Methods  
In total, 76 samples were collected, consisting of milk (\(n\) = 53) and feeds (\(n\) = 23) from Farms A to E (Table 1). Sample collection was performed between June and August 2021, i.e., one year after our previous investigation [2].  
In connection with milk, we collected 5 samples from the farms' raw milk tanks (1 from each of the Farms A-E) and 48 swabs (12 from each farm, excluding Farm B, which applies manual milking) from the interior surface of the shells (teatcups) of each milking machine, after routine disinfection. Four (\(n\) = 4) of the swabs collected from each farm were taken from the bottom of different shells, and the rest (\(n\) = 8) from the area close to the shell's ring.  
With regards to feeds, this investigation was focused to own home-grown feeds stored in each farm's respective storage (\(n\) = 6, 1 in Farms A, B, D, and E, and 2 from Farm C), and to those of the ingredients produced by the respective farms (\(n\) = 17, 3 from Farm A, 1 from Farm B, 6 from Farm C, 3 from Farm D, and 4 from Farm E) (Table 1).  
Samples were processed for the detection of _E. coli_, _S. aureus_, and coagul-negative _Staphylococci_ (CoNS), as well as the assessment of drug resistance, using conventional microbiology and the polymerase chain reaction (PCR), as previously described [2]. In brief, sample processing for the isolation of targeted pathogens was conducted based on standard procedures using a 7% sheep blood agar base (Oxoid Ltd., Basingstoke, UK) for milk samples, MacConkey agar (Oxoid Limited, UK) for milk and feed samples, and Baird-Parker agar (Oxoid Limited, UK) for feed samples. Colonies phenotypically consistent with _Staphylococci_ and _E. coli_ were subcultured on Plate Count Agar (PCA, Oxoid Limited, UK) or Tryptone Soya Agar (TSA, Oxoid Limited, UK) and Tryptone Bile Xylucuronide agar (TBX agar, Oxoid Limited, UK), respectively. Selected colonies were tested with PCR targeting the _tuf_ gene for _Staphylococcus_ spp., the _nuc_ gene for _S. aureus_ and the _rfbE_ gene for _E. coli_[2].  
_Staphylococci_ isolates were tested for susceptibility to cefoxitin (FOX: 30 \(\upmu\)g), clindamycin (DA: 2 \(\upmu\)g), erythromycincin (ERY: 15 \(\upmu\)g), gentamicin (CN: 10 \(\upmu\)g), nonfloxacin (NOR: 10 \(\upmu\)g), penicillin G (P: 1 unit), tetracycline (TE: 30 \(\upmu\)g), and trimethoprim-sulphamethoxazole (SXT: 25 \(\upmu\)g). The isolates of _E. coli_ were tested for susceptibility to amoxicillin-lin-clavulanic acid (AMC: 30 \(\upmu\)g), ampicillin (AM: 10 \(\upmu\)g), cefotaxime (CTX: 5 \(\upmu\)g), cefoxitin (FOX: 30 \(\upmu\)g), ceftazidine (CAZ: 10 \(\upmu\)g), ciprofloxacin (CIP: 5 \(\upmu\)g), gentamicin (CN: 10 \(\upmu\)g), meropenem (MEM: 10 \(\upmu\)g), and trimethoprim-sulphamethoxazole (SXT: 25 \(\upmu\)g).  
Isolates were characterised as AMR if not susceptible to at least one of the antimicrobials recommended by the European Committee on Antimicrobial Susceptibility Testing, at a concentration corresponding to the determined breakpoint [4], and as MDR when non-susceptible to at least one agent from three antimicrobial categories [5].  
\begin{table}
\begin{tabular}{l c c c c c c c c c c c c c c c c c c c c c c c c c c c} \hline \hline  & \multicolumn{4}{c}{**Fam A**} & \multicolumn{4}{c}{**Fam B**} & \multicolumn{4}{c}{**Fam C**} & \multicolumn{4}{c}{**Fam D**} & \multicolumn{4}{c}{**Fam E**} & \multicolumn{4}{c}{**Total**} & \multicolumn{1}{c}{**Sulphyaccool (So)**} & \multicolumn{1}{c}{**Ec**} & \multicolumn{1}{c}{**Surn**} \\
**Type of Sample** & **T