We present a workflow, from sampling to interpretation, showing how resistance monitoring can be carried out in swine herds using a metagenomic approach. We propose metagenomic sequencing should be part of routine livestock resistance monitoring programmes and potentially of integrated One Health monitoring in all reservoirs.

Combination therapy with several antibiotics is one strategy that has been applied in order to limit the spread of antimicrobial resistance. We compared the de novo evolution of resistance during combination therapy with the β-lactam ceftazidime and the fluoroquinolone ciprofloxacin with the resistance evolved after single-drug exposure. Combination therapy selected for mutants that displayed broad-spectrum resistance, and a major resistance mechanism was mutational inactivation of the repressor gene mexR that regulates the multidrug efflux operon mexAB-oprM. Deregulation of this operon led to a broad-spectrum resistance phenotype that decreased susceptibility to the combination of drugs applied during selection as well as to unrelated antibiotic classes. Mutants isolated after single-drug exposure displayed narrow-spectrum resistance and carried mutations in the MexCD-OprJ efflux pump regulator gene nfxB conferring ciprofloxacin resistance, or in the gene encoding the non-essential penicillin-binding protein DacB conferring ceftazidime resistance. Reconstruction of resistance mutations by allelic replacement and in vitro fitness assays revealed that in contrast to single antibiotic use, combination therapy consistently selected for mutants with enhanced fitness expressing broad-spectrum resistance mechanisms.

The tetracycline resistance gene tet(K) was shown to be integrated within the predominant staphylococcal cassette chromosome mec (SCCmec) element of Danish livestock-associated methicillin-resistant Staphylococcus aureus CC398 (LA-MRSA CC398). These LA-MRSA CC398 isolates already possessed tet(M), but the acquisition of tet(K) significantly improved their fitness at sublethal concentrations of tetracycline. Because tet(K) is genetically linked to SCCmec, the use of tetracycline in food animals may have contributed to the successful spread of LA-MRSA CC398.

Crossbreeding in dairy cattle has recently become of increased interest. However, farmers in Scandinavian countries are reluctant to implement crossbreeding in their herds, and one reason is the common opinion that only herds at a poor level of management can benefit from crossbreeding. The Danish Cattle Database (SEGES, Aarhus, Denmark) provided data on 14 traits regarding milk yield, udder health, fertility traits, stillbirth, and survival. The data were collected from 103,307 pure Holstein cows and 14,832 F 1 crosses (Holstein dam and Nordic Red sire). The cows were born between 2008 and 2014 and originated from 424 herds that contributed data from at least 5 purebreds and 5 crossbreds across the years. We split the animals into 3 production levels: high, average, and low according to the herd's average production (kg) of 305-d fat plus protein in the given birth year of the cow. We estimated least squares means of breed group (purebred and crossbred) performance within each production level. Crossbred performance in 305-d fat yield in first-parity cows was greater than that of Holstein across all herd production levels; the gain was greater in high-(9 kg more than Holstein) and average-producing herds (7 kg more than Holstein) than in low-producing herds (3 kg more than Holstein). Regardless of production level or parity, crossbreds did not outperform Holstein in terms of 305-d protein yield (0 to 8 kg less). Crossbreds had relatively better udder health than Holstein in both first and second parity (up to 15% less mastitis) within any of the production levels. In terms of fertility, stillbirth, and survival, crossbreds performed better than purebreds, and improved performance was independent of herd production level. We conclude that differences in performance between F 1 crossbreds and Holstein are independent of production level.

This study simulated the consequences of crossbreeding between Swedish Holstein and Swedish Red on herd dynamics and herd profitability under Swedish conditions. Two base herds were simulated using a stochastic herd simulation model, SimHerd Crossbred. The herds reflected average Swedish conventional and organic herds having purebred Swedish Holstein. For each base herd, 3 breeding strategies were simulated: purebreeding, 2-breed terminal crossbreeding, and 2-breed rotational crossbreeding. The terminal crossbreeding strategy implied having a nucleus of Swedish Holstein and a proportion of F 1 Swedish Red × Swedish Holstein crossbred cows within the same herd. The crossbreds in this herd did not produce replacement heifers but exclusively beef × dairy cross calves. Beef semen was also used in the pure-breeding (10-20% in cows) and the rotational crossbreeding (40% in cows) strategies to retain a limited surplus of replacement heifers. To ensure an adequate number of crossbreds in the terminal crossbreeding strategy, X-sorted sexed semen was used for insemination in all the purebred heifers. The outcome was 67% purebred and 31% F 1 crossbreds in the herd. In addition, 31% heterosis was expressed compared with 67% heterosis expressed using a 2-breed rotational crossbreeding strategy. Compared with the pure-breeding strategy, crossbreeding increased the annual contribution margin per cow by €20 to €59, with the rotational crossbreeding strategy creating the largest profitability. The increased profitability was mainly due to improved functional traits, especially fertility. For the conventional production system, the replacement rate was 39.3% in the pure-breeding strategy and decreased to 35.8 and 30.1% in the terminal and rotational crossbreeding strategy, respectively. Similar changes happened in the organic production system. Additionally, the crossbreeding strategies earned €22 to €42 more annually per cow from selling live calves for slaughter due to the extended use of beef semen. Milk production was similar between pure-breeding and terminal crossbreeding, and only decreased 1 to 2% in rotational crossbreeding. These results show that crossbreeding between Swedish Holstein and Swedish Red can be profitable in both conventional and organic Swedish herds using the strategies we have simulated. However, some aspects remain to be investigated, such as the economically optimal breeding strategy, genetic improvement, and transition strategies.