Nucleic acid separation is an increasingly important tool for molecular biology. Before modern technologies could be used, nucleic acid separation had been a time- and work-consuming process based on several extraction and centrifugation steps, often limited by small yields and low purities of the separation products, and not suited for automation and up-scaling. During the last few years, specifically functionalised magnetic particles were developed. Together with an appropriate buffer system, they allow for the quick and efficient purification directly after their extraction from crude cell extracts. Centrifugation steps were avoided. In addition, the new approach provided for an easy automation of the entire process and the isolation of nucleic acids from larger sample volumes. This review describes traditional methods and methods based on magnetic particles for nucleic acid purification. The synthesis of a variety of magnetic particles is presented in more detail. Various suppliers of magnetic particles for nucleic acid separation as well as suppliers offering particle-based kits for a variety of different sample materials are listed. Furthermore, commercially available manual magnetic separators and automated systems for magnetic particle handling and liquid handling are mentioned.

Magnetic particles for separation and purification of nucleic acids

Antibiotics are widely used in animal husbandry, and various types of antibiotic resistance genes (ARGs) are frequently detected in livestock waste around the world. Conventional livestock waste treatment processes do not completely remove ARGs, resulting in their release to soil and water environments. Various exposure routes of these ARGs to humans, including inhalation and ingestion of antibiotic-resistant bacteria (ARB) that harbor them, may be contributing to the rise in resistant clinical infections that are increasingly difficult to treat with antibiotics. In this review, we assess the occurrence and variability of ARGs in livestock wastes and their potential propagation pathways to human pathogens. We also review the mechanisms and environmental factors that influence the dissemination of ARGs through these pathways, and evaluate the ARG removal efficiency of common livestock waste management approaches. Challenges and research needs for assessing and mitigating the risk of antibiotic resistance dissemination from livestock waste are also presented.

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Antibiotic resistance genes from livestock waste: occurrence, dissemination, and treatment

Intracellular versus extracellular antibiotic resistance genes in the environment: Prevalence, horizontal transfer, and mitigation strategies.

There is a growing need to mitigate the discharge of extracellular antibiotic resistance genes (ARGs) from municipal wastewater treatment systems. Here, molecularly-imprinted graphitic carbon nitride (MIP-C3N4) nanosheets were synthesized for selective photocatalytic degradation of a plasmid-encoded ARG (blaNDM-1, coding for multidrug resistance New Delhi metallo-β-lactamase-1) in secondary effluent. Molecular imprinting with guanine enhanced ARG adsorption, which improved the utilization of photogenerated oxidizing species to degrade blaNDM-1 rather than being scavenged by background nontarget constituents. Consequently, photocatalytic removal of blaNDM-1 in secondary effluent with MIP-C3N4 (k = 0.111 ± 0.028 min-1) was 37 times faster than with bare graphitic carbon nitride (k = 0.003 ± 0.001 min-1) under UVA irradiation (365 nm, 3.64 × 10-6 Einstein/L·s). MIP-C3N4 can efficiently catalyze the fragmentation of blaNDM-1, which decreased the potential for ARG repair by transformed bacteria. Molecular imprinting also changed the primary degradation pathway; electron holes (h+) were the predominant oxidizing species responsible for blaNDM-1 removal with MIP-C3N4 versus free radicals (i.e., ·OH and O2-) for coated but nonimprinted C3N4. Overall, MIP-C3N4 efficiently removed blaNDM-1 from secondary effluent, demonstrating the potential for molecular imprinting to enhance the selectivity and efficacy of photocatalytic processes to mitigate dissemination of antibiotic resistance from sewage treatment systems.

Selective Adsorption and Photocatalytic Degradation of Extracellular Antibiotic Resistance Genes by Molecularly-Imprinted Graphitic Carbon Nitride.

Quantifying the occurrence and transformation potential of extracellular polymeric substances (EPS)-associated antibiotic resistance genes in activated sludge.

Redistribution of intracellular and extracellular free & adsorbed antibiotic resistance genes through a wastewater treatment plant by an enhanced extracellular DNA extraction method with magnetic beads

Qing Bin Yuan

Papers

Redistribution of intracellular and extracellular free & adsorbed antibiotic resistance genes through a wastewater treatment plant by an enhanced extracellular DNA extraction method with magnetic beads