Leaching of antibiotic resistance genes and microbial assemblages following poultry litter applications in karst and non-karst landscapes.

Antibiotic resistance is increasingly recognized as a critical challenge affecting human, animal, and environmental health. Yet, environmental dynamics and transport of antibiotic resistance genes (ARGs) and microbial communities in karst and non-karst leachate following poultry litter land applications are not well understood. This study investigates impacts of broiler poultry litter application on the proliferation of ARGs (tetW, qnrS, ermB, sulI, and blaCTX-M-32), class 1 integron (intI1 i), and alterations in microbial communities (16S rRNA) within karst derived soils, which are crucial and under-researched systems in the global hydrological cycle, and non-karst landscapes. Using large, intact soil columns (45 cm diam. × 100 cm depth) from karst and non-karst landscapes, the role of preferential flow and ARG transport in leachate was enumerated following surface application of poultry litter and simulated rain events. This research demonstrated that in poultry litter amended karst soils, ARG (i.e., ermB and tetW) abundance in leachate increased 1.5 times compared to non-karst systems (p < 0.05), highlighting the influence of geological factors on ARG proliferation. Notably, microbial communities in karst soil leachate exhibited increased diversity and abundance, suggesting a potential linkage between microbial composition and ARG presence. Further, our correlation and network analyses identified relationships between leachate ARGs, microbial taxa, and physicochemical properties, underscoring the complex interplay in these environmentally sensitive areas. These findings illuminate the critical role of karst systems in shaping ARG abundance and pollutant dispersal and microbial community dynamics, thus emphasizing the need for landscape-specific approaches in managing ARG dissemination to the environment. This study provides a deeper understanding of hydrogeological ARG dynamics but also lays the groundwork for future research and strategies to mitigate ARG dissemination through targeted manure applications across agricultural landscapes.

Long-term impacts of conservation pasture management in manuresheds on system-level microbiome and antibiotic resistance genes.

Animal manure improves soil fertility and organic carbon, but long-term deposition may contribute to antibiotic resistance genes (ARGs) entering the soil-water environment. Additionally, long-term impacts of applying animal manure to soil on the soil-water microbiome, a crucial factor in soil health and fertility, are not well understood. The aim of this study is to assess: (1) impacts of long-term conservation practices on the distribution of ARGs and microbial dynamics in soil, and runoff; and (2) associations between bacterial taxa, heavy metals, soil health indicators, and ARGs in manures, soils, and surface runoff in a study following 15 years of continuous management. This management strategy consists of two conventional and three conservation systems, all receiving annual poultry litter. High throughput sequencing of the 16S ribosomal RNA was carried out on samples of cattle manure, poultry litter, soil, and runoff collected from each manureshed. In addition, four representative ARGs (intl1, sul1, ermB, and blactx-m-32) were quantified from manures, soil, and runoff using quantitative PCR. Results revealed that conventional practice increased soil ARGs, and microbial diversity compared to conservation systems. Further, ARGs were strongly correlated with each other in cattle manure and soil, but not in runoff. After 15-years of conservation practices, relationships existed between heavy metals and ARGs. In the soil, Cu, Fe and Mn were positively linked to intl1, sul1, and ermB, but trends varied in runoff. These findings were further supported by network analyses that indicated complex co-occurrence patterns between bacteria taxa, ARGs, and physicochemical parameters. Overall, this study provides system-level linkages of microbial communities, ARGs, and physicochemical conditions based on long-term conservation practices at the soil-water-animal nexus.

Antibiotic resistance in soil and tomato crop irrigated with freshwater and two types of treated wastewater.

Agricultural use of treated wastewater (TWW) is an effective means to reduce freshwater (FW) consumption. However, there is a growing concern regarding the potential dissemination of antibiotic resistance elements by TWW irrigation. We hypothesized that higher levels of antibiotic resistance genes (ARGs) would be detected in soil and crops irrigated with TWW compared to FW irrigation. To test our prediction, samples of water (FW, secondary TWW, and tertiary TWW), irrigated soils, and crops (tomato) surface wash were collected during two consecutive growing seasons. The ARGs conferring resistance to sulfonamide, fluoroquinolone, penicillin, erythromycin and tetracycline were quantified in the samples, alongside Class 1 integron-integrase and the bacterial 16 S rRNA encoding genes. Contrary to our hypothesis, ARGs in the irrigation water were not propagated to either the irrigated soil, or the tomato. The tomato surface wash featured a variety of ARGs that were undetected in neither the waters nor the irrigated soils. Therefore, we cautiously question the link between irrigation water quality and the soil and produce resistomes.

The dissemination of antibiotics and their corresponding resistance genes in treated effluent-soil-crops continuum, and the effect of barriers.

Irrigation with treated effluent is expanding as freshwater sources diminish, but hampered by growing concerns of pharmaceuticals contamination, specifically antibiotics and resistance determinants. To evaluate this concern, freshwater and effluent were applied to an open field that was treated with soil barriers including plastic mulch together with surface and subsurface drip irrigation, cultivating freshly eaten crops (cucumbers or melons) for two consecutive growing seasons. We hypothesized that the effluent carries antibiotics and resistance determinants to the drip-irrigated soil and crops regardless of the treatment. To test our hypothesis, we monitored for antibiotics abundance (erythromycin, sulfamethoxazole, tetracycline, chlortetracycline, oxytetracycline, amoxicillin, and ofloxacin) and their corresponding resistance genes (ermB, ermF, sul1, tetW, tetO, blaTEM and qnrB), together with class 1 integron (intl1), and bacterial 16S rRNA, in water, soil, and crop samples taken over two years of cultivation. The results showed that an array of antibiotics and their corresponding resistance genes were detected in the effluent but not the freshwater. Yet, there were no significant differences in the distribution or abundance of antibiotics and resistance genes, regardless of the irrigation water quality, or crop type (p > 0.05), but plastic-covered soil irrigated with effluent retained the antibiotics oxytetracycline and ofloxacin (p < 0.05). However, we could not detect significant correlations between the detected antibiotics and the corresponding resistance genes. Overall, our findings disproved our hypothesis suggesting that treated effluent may not carry antibiotics resistance genes to the irrigated soil and crops yet, plastic mulch covered soil retain some antibiotics that may inflict long term contamination.

Occurrence and distribution of antibiotics and corresponding antibiotic resistance genes in different soil types irrigated with treated wastewater.

Diminishing freshwater (FW) supplies necessitate the reuse of treated wastewater (TWW) for various purposes, like irrigation of agricultural lands. However, there is a growing concern that irrigation with TWW may transfer antibiotic resistance genes (ARGs) to the soil and crops. We hypothesized that TWW irrigation would increase the prevalence of antibiotic residues together with the corresponding ARGs in the irrigated soil. We further predicted that soil texture, especially pH, clay content, and organic matter variabilities, would change the antibiotic residues concentrations and thus ARGs dissemination. To test our predictions, three soils types (loamy-sand, loam, and clay) were irrigated with two water types (FW and TWW), over two consecutive seasons. We monitored physico-chemical parameters, the abundance of seven antibiotic residues, and their corresponding ARGs together with class 1 integron (intI1) in 54 water and soil samples collected at the end of the field experiments. The results revealed increase in antibiotics concentrations and ARGs relative abundance in TWW than FW. Yet, in the soil ARGs relative abundances were independent of the irrigation water quality, but dependent on the soil type, especially the clay content. Further, there were no clear associations between the targeted antibiotics or the presence of heavy metals and ARGs' relative abundance in the water or soil samples. Therefore, our results question the link between the discharge of antibiotics and heavy metals, and the dissemination of ARGs in soil environments.