Since the COVID-19 pandemic started, there has been much speculation about how COVID-19 and antimicrobial resistance may be interconnected. In this study, untreated wastewater was sampled from Hospital A designated to treat COVID-19 patients during the first wave of the COVID-19 pandemic alongside Hospital B that did not receive any COVID-19 patients. Metagenomics was used to determine the relative abundance and mobile potential of antibiotic resistant genes (ARGs), prior to determining the correlation of ARGs with time/incidence of COVID-19. Our findings showed that ARGs resistant to macrolides, sulfonamides, and tetracyclines were positively correlated with time in Hospital A but not in Hospital B. Likewise, minor extended spectrum beta-lactamases (ESBLs) and carbapenemases of classes B and D were positively correlated with time, suggesting the selection of rare and/or carbapenem-resistant genes in Hospital A. Non-carbapenemase bla_VEB also positively correlated with both time and intI1 and was copresent with other ARGs including carbapenem-resistant genes in 6 metagenome-assembled genomes (MAGs). This study highlighted concerns related to the dissemination of antimicrobial resistance (AMR) during the COVID-19 pandemic that may arise from antibiotic use and untreated hospital wastewater.

The role of sorption and biodegradation in a membrane aerated biofilm reactor (MABR) were investigated for the removal of 10 organic micropollutants (OMPs) including endocrine disruptors and pharmaceutical active compounds. The influence of the biofilm thickness on the mechanisms of removal was analyzed via kinetic test at three different stages. At all biofilm stages, biodegradation was demonstrated to dominate the removal of selected OMPs. Higher OMPs rates of removal via biodegradation (K_biol) were achieved when biofilm increased its thickness from (stage T1) 0.26 mm, to (stage T2) 0.58 mm and (stage T3) 1.03 mm. At stage T1 of biofilm, heterotrophs contribute predominantly to OMPs degradation. Hydrophilic compounds removal (i.e., acetaminophen) continue to be driven by heterotrophic bacteria at the next stages of biofilm thickness. However, for medium hydrophobic neutral and charged OMPs, the combined action of heterotrophic and enriched nitrifying activity at stages T2 and T3 enhanced the overall removal. A degradation pathway based on heterotrophic activity for acetaminophen and combined action of nitrifiers-heterotrophs for estrone was proposed based on identified metabolites. Although biodegradation dominated the removal of most OMPs, sorption was also observed to be essential in the removal of biologically recalcitrant and lipophilic compounds like triclosan. Furthermore, sorption capacity of apolar compound was enhanced as the biofilm thickness grew and increased in EPS protein fraction. Microbial analysis confirmed the higher abundance of nitrifying and denitrifying activity at stage T3 of biofilm, which not only facilitated near complete ammonium removal but also enhanced degradation of OMPs.

The types and proportions of soluble sugar and organic acid in fruit significantly affect flavor quality. However, there are few reports on the crosstalk regulation between metabolism of organic acid and sugar in fruit. Here, we found that the overexpression of cytoplasmic malate dehydrogenase genes (MdcyMDHs) not only increased the malate content but also increased the sucrose concentration in transgenic apple calli and mature fruit. Enzyme activity assays indicated that the overexpression of MdcyMDH1 and MdcyMDH5 enhanced sucrose phosphate synthase (SPS) activity in transgenic materials. RNA-seq and expression analysis showed that the expression levels of SPS genes were up-regulated in MdcyMDH1-overexpressed apple fruit and MdcyMDH5-overexpressed apple calli. Further study showed that the inhibition of MdSPSB2 or MdSPSC2 expression in MdcyMDH1 transgenic fruit could reduce or eliminate, respectively, the positive effect of MdcyMDH1 on sucrose accumulation. Moreover, some starch cleavage-related genes (MdBAM6.1/6.2, MdBMY8.1/8.2, MdISA1) and the key gluconeogenesis-related phosphoenolpyruvate carboxykinase MdPEPCK1 gene were significantly up-regulated in the transcriptome differentially expressed genes of mature fruit overexpressing MdcyMDH1. These results indicate that alteration of malate metabolism mediated by MdcyMDH might regulate the expression of MdSPSs and SPS activity via affecting starch metabolism or gluconeogenesis, and thus accelerate sucrose synthesis and accumulation in fruit.

Mass gatherings (MGs), such as Hajj and Umrah, gather vast and diverse populations and potentially contribute to antimicrobial resistance (AMR) in the host country. However, assessing this impact is challenging due to the lack of a suitable baseline. During the COVID-19 pandemic restrictions and lifting, 185 sewage samples were collected from four wastewater treatment plants affected by MG and a control wastewater treatment plant, covering 3 Hajj seasons and 2 Ramadan periods. Here we show that despite no overall increase in total antibiotic resistance gene level, metagenomic analysis revealed that MGs introduced antibiotic resistance genes, including bla_PER, into the local sewage. This gene was also found in a genomic island facilitated by the IS26 array, which was identified in viable opportunistic pathogens, including Shewanella putrefaciens, Shewanella xiamenesis and Aeromonas media, indicating emerging AMR threats to the local environment. These insights are crucial for informing public health strategies and interventions, ensuring better preparedness and response to AMR dissemination during MGs.

Sludge is a biohazardous solid waste that is produced during wastewater treatment. It contains antibiotic resistance genes (ARGs) that pose significant antimicrobial resistance (AMR) threats. Herein, aerobic and anaerobic membrane bioreactors (AeMBRs and AnMBRs, respectively) were compared in terms of the volume of waste sludge generated by them, the presence of ARGs in the sludge, and the potential for horizontal gene transfer (HGT) events using metagenomics to determine which treatment process can better address AMR concerns associated with the generation of waste sludge. The estimated abundance of ARGs in the suspended sludge generated by the AnMBR per treated volume is, on average, 5-55 times lower than that of sludge generated by the AeMBR. Additionally, the ratio of potential HGT in the two independent runs was lower in the anaerobic sludge (0.6 and 0.9) compared with that in the aerobic sludge (2.4 and 1.6). The AnMBR sludge exhibited reduced HGT of ARGs involving potential opportunistic pathogens (0.09) compared with the AeMBR sludge (0.27). Conversely, the AeMBR sludge displayed higher diversity and more transfer events, encompassing genes that confer resistance to quinolones, rifamycin, multidrug, aminoglycosides, and tetracycline. A significant portion of these ARGs were transferred to Burkholderia sp. By contrast, the AnMBR showed a lower abundance of mobile genetic elements associated with conjugation and exhibited less favorable conditions for natural transformation. Our findings suggest that the risk of potential HGT to opportunistic pathogens is greater in the AeMBR sludge than in AnMBR sludge.

Water reuse is increasingly pursued to alleviate global water scarcity. However, the wastewater treatment process does not achieve full removal of biological contaminants from wastewater, hence microorganisms and their genetic elements can be disseminated into the reclaimed water distribution systems (RWDS). In this study, reclaimed water samples are investigated via metagenomics to assess their bacterial diversity, metagenome-assembled genomes (MAGs) and antibiotic resistance genes (ARGs) at both point of entry (POE) and point of use (POU) in 3 RWDS. The number of shared bacterial orders identified by metagenome was higher at the POE than POU among the three sites, indicating that specific conditions in RWDS can cause further differentiation in the microbial communities at the end of the distribution system. Two bacterial orders, namely Rhizobiales and Sphingomonadales, had high replication rates in two of the examined RWDS (i.e., site A and B), and were present in higher relative abundance in POU than at POE. In addition, MAG and ARG relative abundance exhibited a strong correlation (R² = 0.58) in POU, indicating that bacteria present in POU may have a high incidence of ARG. Specifically, resistance genes associated with efflux pump mechanisms (e.g., adeF and qacH) increased in its relative abundance from POU to POE at two of the RWDS (i.e., site A and B). When correlated with the water quality data that suggests a significantly lower dissolved organic carbon (DOC) concentration at site D than the other two RWDS, the metagenomic data suggest that low DOC is needed to maintain the biological stability of reclaimed water along the distribution network.