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The spread of antibiotic resistance is closely related with selective pressure in the environment. Wastewater from industrialized regions is characterized by higher concentrations of these pollutants than sewage from less industrialized areas. The aim of this study was to compare the concentrations of contaminants such as antibiotics and heavy metals (HMs), and to evaluate their impact on the spread of genes encoding resistance to antimicrobial drugs in samples of wastewater, sewage sludge and river water in two regions with different levels of industrialization. The factors exerting selective pressure, which significantly contributed to the occurrence of the examined antibiotic resistance genes (ARGs), were identified. The concentrations of selected gene copy numbers conferring resistance to four groups of antibiotics as well as class 1 and 2 integron-integrase genes were determined in the analyzed samples. The concentrations of six HMs and antibiotics corresponding to genes mediated resistance from 3 classes were determined. Based on network analysis, only some of the analyzed antibiotics correlated with ARGs, while HM levels were correlated with ARG concentrations, which can confirm the important role of HMs in promoting drug resistance. The samples from a wastewater treatment plant (WWTP) located an industrialized region were characterized by higher HM contamination and a higher number of significant correlations between the analyzed variables than the samples collected from a WWTP located in a less industrialized region. These results indicated that treated wastewater released into the natural environment can pose a continuous threat to human health by transferring ARGs, antibiotics and HMs to the environment. These findings shed light on the impact of industrialization on antibiotic resistance dissemination.

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The starch content of triticale and oat grains provides much of their readily available energy. Synchronizing energy and nitrogen in the rumen is important in optimizing profitability; for this reason, ammonia processing of these grains was evaluated for its potential to modify ruminal fermentation and to improve milk production performance. A mixture of ground triticale and oats (CONG, in a 60:40 ratio 40 by DM) was treated with urea (5 kg/1000 kg) and urease additive (20 kg/1000 kg) containing 200 g/kg of moisture, for 2 wk (UREG). The urea treatment enhanced the pH and CP content of grains by 34% and 52%, respectively. In a batch culture study, CONG or UREG as the only substrate was incubated in a buffered ruminal fluid. Compared to CONG, UREG increased pH, total VFA concentration, total gas, and disappearance of DM, while reducing CH4 production, whereas NH3 concentration increased and entodiniomorph counts tended to increase. In the in vivo study, cows were randomly allocated to two dietary groups (n = 24) and were offered TMR based on maize and grass silage, containing either 155 g/kg of CONG and 80 g/kg of soybean meal (CONT) or 155 g/kg of UREG and 59 g/kg of soybean meal (URET) for 31 d. Ruminal fluid was collected (n = 10) using rumenocentesis. The relative abundances of Streptococcus bovis decreased, but Megasphaera elsdenii, methanogens, and ammonia-producing bacteria increased by URET. Entodiniomorph and holotrich counts were decreased by URET. Feeding with URET increased ruminal pH and concentrations of total VFA, acetate, branched-chain VFA, and NH3. Feeding with URET also increased milk yield. These results demonstrate that replacing untreated triticale and oat grains with urea-treated grains can beneficially modulate ruminal microbiota and fermentation, consequently improving production performance and profitability.

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