Antimicrobial resistance (AMR) is a growing threat to human and animal health.Progress in molecular biology has revealed new and significant challenges for AMR mitigation given the immense diversity of antibiotic resistance genes (ARGs), the complexity of ARG transfer, and the broad range of omnipresent factors contributing to AMR. Municipal, hospital and abattoir wastewater are collected and treated in wastewater treatment plants (WWTPs), where the presence of diverse selection pressures together with a highly concentrated consortium of pathogenic/commensal microbes create favourable conditions for the transfer of ARGs and proliferation of antibiotic resistant bacteria (ARBs). The recent emergence ARBs and ARGs as well as their potential health effects have re-defined the role of WWTPs as a focal point in the fight against AMR. By reviewing the occurrence of ARGs in wastewater and sludge and the current technologies used to quantify ARGs and identify antibiotic resistant bacteria (ARB), this paper provides a research roadmap to address existing challenges in AMR control via wastewater treatment. Wastewater treatment is a double-edged sword that can act as either a pathway for AMR spread or as a barrier to reduce the environmental release of anthropogenic AMR. State of the art ARB identification technologies, such as metagenomic sequencing and fluorescence-activated cell sorting, have enriched ARG/ARB databases, unveiled keystone species in AMR networks, and improved the resolution of AMR dissemination models. Data and information provided in this review highlight significant knowledge gaps. These include inconsistencies in ARG reporting units, lack of ARG/ARB monitoring surrogates, lack of a standardised protocol for determining ARG removal via wastewater treatments, and the inability to support appropriate risk assessment. This is due to a lack of standard monitoring targets and agreed threshold values, and paucity of information on the ARG-pathogen host relationship and potential risk evolution. These research gaps need to be addressed and research 3 findings need to be transformed into practical guidance for WWTP operators to enable effective progress towards mitigating the evolution and spread of AMR.

This study investigated the production of biogas, volatile fatty acids (VFAs), and other soluble organic from lignocellulosic biomass by two microbial communities (i.e. rumen fluid and anaerobic sludge). Four types of abundant lignocellulosic biomass (i.e. wheat straw, oaten hay, lurence hay and corn silage) found in Australia were used. The results show that rumen microbes produced four-time higher VFAs level than that of anaerobic sludge reactors, indicating the possible application of rumen microorganism for VFAs generation from lignocellulosic biomass. VFA production in the rumen fluid reactors was probably due to the presence of specific hydrolytic and acidogenic bacteria (e.g. Fibrobacter and Prevotella).VFA production corroborated from the observation of pH drop in the rumen fluid reactors indicated hydrolytic and acidogenic inhibition, suggesting the continuous extraction of VFAs from the reactor. Anaerobic sludge reactors on the other hand, produced more biogas than that of rumen fluid reactors. This observation was consistent with the abundance of methanogens in anaerobic sludge inoculum (3.98% of total microbes) compared to rumen fluid (0.11%). VFA production from lignocellulosic biomass is the building block chemical for bioplastic, biohydrogen and biofuel. The results from this study provide important foundation for the development of engineered systems to generate VFAs from lignocellulosic biomass.

The removal efficiency of 22 selected trace organic contaminants by sequential application of granular activated carbon (GAC) and simultaneous application of powdered activated carbon (PAC) with membrane bioreactor (MBR) was compared in this study. Both sequential application of GAC following MBR treatment (MBR-GAC) and simultaneous application of PAC within MBR (PAC-MBR) achieved improved removal (over 95%) of seven hydrophilic and biologically persistent compounds, which were less efficiently removed by MBR-only treatment (negligible to 70%). However, gradual breakthrough of these compounds occurred over an extended operation period. Charged compounds, particularly, fenoprop and diclofenac, demonstrated the fastest breakthrough (complete and 50-70%, in MBR-GAC and PAC-MBR, respectively). Based on a simple comparison from the long-term performance stability and activated carbon usage points of view, PAC-MBR appears to be a better option than MBR-GAC treatment.

Effects of operational disturbance and subsequent recovery process on microbial community during a pilot-scale anaerobic co-digestion

BackgroundDengue is a rapidly spreading mosquito borne tropical viral disease affecting hundreds of millions of people across the globe annually. The dengue virus (DENV) includes four genetically distinct serotypes that cause serious life-threatening infections, including dengue hemorrhagic fever/dengue shock syndrome. Dengue vaccine development is complicated by the possibility of vaccine-enhanced severe dengue disease due to antibody-dependent enhancement by pre-existing cross-reactivity, as well as homotypic antibodies. Thus, the development of an efficacious dengue vaccine conferring simultaneous and durable immunity to each of the four DENV serotypes has not yet been developed despite years of research. For mass immunization in deeply affected resource-limited countries, oral vaccination is considered more beneficial than conventional approaches. Therefore, in a continuing effort towards designing economical and potent vaccine candidates, the current study applied yeast surface display technology to develop an oral dengue vaccine candidate using whole recombinant yeast cells displaying the recombinant fusion protein of M cell targeting ligand Co1 fused to the synthetic consensus dengue envelope domain III (scEDIII). Female Balb/c mice were orally fed with recombinant yeast cells and immunogenicity in terms of systemic and mucosal immune responses was monitored.ResultsImmunofluorescence microscopy with dengue specific antibody and fluorescein isothiocyanate-conjugated anti-mouse IgG antibody clearly showed that recombinant protein Co1-scEDIII-AGA was localized on the cell surface of the respective clones in comparison with scEDIII-Co1 and Mock cells with no fluorescence. Oral dosage applications of surface displayed Co1-scEDIII-AGA stimulated a systemic humoral immune response in the form of dengue-specific serum IgG, as well as a mucosal immune response in the form of secretory immunoglobulin A (sIgA). Antigen-specific B cell responses in isolated lymphoid cells from the spleen and Peyer’s patches further supported an elevated mucosal immune response. In addition, surface displayed Co1-scEDIII-AGA feeding elicited strong immune responses in comparison with scEDIII-Co1 and Mock following intraperitoneal booster with purified scEDIII antigen.ConclusionsSurface displayed preparations of Co1-scEDIII-AGA induced strong immunogenicity compared with non-displayed scEDIII-Co1. Prior studies have supported the neutralization potential of scEDIII constructs against all four serotypes. Thus, the oral administration of genetically engineered yeast whole cells displaying biologically active Co1-scEDIII fusion protein without any further processing shows prospective as a potent oral vaccine candidate against dengue viral infection.

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