Fate of sulfadiazine administered to pigs and its quantitative effect on the dynamics of bacterial resistance genes in manure and manured soil

The veterinary antibiotic sulfadiazine (SDZ), labelled by 14C, was administered to pigs to follow the fate of the drug and its metabolites in manure and manure-amended soil, and to investigate the dynamics of drug effects on resistance genes and bacterial communities. In the manure sampled over 10 days, more than 96% of the drug was found as parent compound or metabolites N-acetyl-SDZ and 4-hydroxy-SDZ. While the manure was stored the concentration of SDZ increased by 42% due to deacetylation of the metabolite N-acetyl-SDZ, whereas the minor metabolite 4-hydroxy-SDZ kept constant. In the soil the extractable amounts of the compounds decreased exponentially to less than 1 mg kg¹ within 11 days after manure amendment. The abundances of SDZ resistance genes sul1 and sul2 were determined by qPCR relative to 16S rRNA genes in total DNA from manure and manure-amended soil. In manure both genes increased exponentially in copy number during the first 60 days of storage, suggesting preferential growth of resistant populations. However, the abundance of sul1 and sul2 decreased below 10⁵ copies per 16S rRNA gene after 175 days. With manure high amounts of sul1 and sul2 were introduced into the soil which were reduced by more than 10 times within 24 days. Thereafter, sul1 was stably maintained in soil, while sul2 further decreased between day 60 and day 165. A mathematical model was developed that could well explain the time course of sul gene abundance by considering the cost of sul genes, horizontal gene transfer, and selection of the resistant populations in the presence of SDZ. Modelling revealed a selective effect of SDZ on sul2 even at low concentrations down to 0.15 mg kg¹ soil. Bacterial community profiles of manure and manure-amended soil were distinct, indicating that bacteria introduced with manure do not become prominent in soil. The composition of the bacterial community in manure constantly changed during storage, but mainly during the first 10 days. Profiles of soil bacterial communities revealed only a transient perturbation by manure containing SDZ.