This study examined differences in antibiotic-resistant soil bacteria and the presence and quantity of resistance genes in soils with a range of management histories. We analyzed four soils from agricultural systems that were amended with manure from animals treated with erythromycin and exposed to streptomycin and/or oxytetracycline, as well as non-manure-amended compost and forest soil. Low concentrations of certain antibiotic resistance genes were detected using multiplex quantitative realtime PCR (qPCR), with tet(B), aad(A), and str(A) each present in only one soil and tet(M) and tet(W) detected in all soils. The most frequently detected resistance genes were tet(B), tet(D), tet(O), tet(T), and tet(W) for tetracycline resistance, str(A), str(B), and aac for streptomycin resistance, and erm(C), erm(V), erm(X), msr(A), ole(B), and vga for erythromycin resistance. Transposon genes specific for Tn916, Tn1549, TnB1230, Tn4451, and Tn5397 were detected in soil bacterial isolates. The MIC ranges of isolated bacteria for tetracycline, streptomycin, and erythromycin were 8 to >256 g/ml, 6 to >1,024 g/ml, and 0.094 to >256 g/ml, respectively. Based on 16S rRNA gene similarity, isolated bacteria showed high sequence identity to genera typical of soil communities. Bacteria with the highest MICs were detected in manure-amended soils or soils from agricultural systems with a history of antibiotic use. Non-manure-amended soils yielded larger proportions of antibiotic-resistant bacteria, but these had lower MICs, carried fewer antibiotic resistance genes, and did not display multidrug resistance (MDR).

This study examined the effects of tetracycline and streptomycin on microorganisms in three different soil habitats: forest soil, agricultural soil, and compost. These antibiotics are commonly used in both medical and veterinary therapy as well as in the production of plant biomass and until quite recently, the production of animal biomass. Microcosms were used as model systems in which the number of microorganisms in environments containing different amounts of antibiotics was analyzed. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of tetracycline and streptomycin were determined. The MIC and MBC values of tetracycline against the tested strains were 20 to 180 microg/ mL and 30 to 300 microg/mL, respectively, and of streptomycin, 360 to 500 microg/mL and > or =500 microg/mL, respectively. Resistant bacterial strains were identified and their physiological profiles assessed. Streptomycin and tetracycline were found to reduce the number of bacteria in the studied soils by between 50 and 80%. Soil bacteria were found to be more resistant to streptomycin than to tetracycline. The bacterial species showing the highest resistance to tetracycline were Rhizobium radiobacter, Burkholderia cepacia, Brevundimonas vesicularis, and Pasteurella multocida. Most soils with high concentrations of streptomycin (5 mg/kg) contained Rhizobium radiobacter, Burkholderia cepacia, and Sphingomonas multivorum, among others. The strains most resistant to tetracycline were isolated from agricultural soil that is constantly subjected to tetracycline pressure from animal manures and biosolids. Among resistant strains, opportunistic pathogens were identified.

The present study collected bacterial samples from water and bottom sediments from fish farms located in a nature reserve area in Poland with no recorded history of antibiotic use. The aim of the study was to determine the initial states of tetracycline, streptomycin, and erythromycin resistance before a potential increase of intensive aquaculture and application of antimicrobial agents in that region. With this in mind, the diversity and antibiotic resistance phenotypes and genotypes of isolates from the bottom sediments and water in five of the 13 fish ponds in Raszyn were evaluated. A total of 58 (sediment, n = 24; water, n = 34) non-repetitive and non-susceptible isolates were affiliated to 14 genera. Among the sediment isolates, Pseudomonas spp. and Bacillus spp. were isolated most frequently, and from the water, Stenotrophomonas spp. and Pseudomonas spp. Phenotypically resistant isolates selected by disk diffusion were further screened by polymerase chain reaction (PCR) and amplicon sequencing. The isolates derived from the water showed a greater percentage of phenotypically resistant isolates to each of the three antibiotics. The most common tetracycline resistant genes detected in isolates from both the water and sediment were tet(A), tet(T), tet(W), and tet(34). On the other hand, the genes tet(X), tet(H), tet(M), and tet(BP) were the most frequent among sedimentary isolates, while tet(B), tet(C), tet (D), and tet(32) were prevalent in aquatic isolates. The most prevalent streptomycin resistance genes among the aquatic isolates were aac(6')-I, str(A), and str(B). The erythromycin resistance genes detected in all isolates included msr(A), erm(X), erm(V), erm(F), and erm(E).

The utilization of pig manure as a source of nutrients for the dissimilatory reduction of sulfates present in phosphogypsum was investigated. In both types of media used (synthetic medium and raw pig manure) increased utilization of sulfates with growing COD/SO4(2-)ratio in the medium was observed. The percent of sulfate reduction obtained in synthetic medium was from 18 to 99%, whereas the value for cultures set up in raw liquid manure was from 12% (at COD/SO4(2-) of 0.3) up to as high as 98% (at COD/SO4(2-) equal 3.80). Even with almost complete reduction of sulfates the percent of COD reduction did not exceed 55%. Based on the results obtained it was concluded that the effectiveness of removal of sulfates and organic matter by sulfate-reducing bacteria (SRB) depends to a considerable degree on the proportion between organic matter and sulfates in the purified wastewaters. The optimal COD/SO4(2-)ratio for the removal oforganic matter was between 0.6 and 1.2 whereas the optimal ratio for the removal of sulfates was between 2.4 and 4.8.