One-year stable pilot-scale operation demonstrates high flexibility of mainstream anammox application.

Mainstream nitrogen removal via anammox is widely recognized as a promising wastewater treatment process. However, its application is challenging at large scale due to unstable suppression of nitrite-oxidizing bacteria (NOB). In this study, a pilot-scale mainstream anammox process was implemented in an Integrated Fixed-film Activated Sludge (IFAS) configuration. Stable operation with robust NOB suppression was maintained for over one year. This was achieved through integration of three key control strategies: i) low dissolved oxygen (DO = 0.4 ± 0.2 mg O2/L), ii) regular free nitrous acid (FNA)-based sludge treatment, and iii) residual ammonium concentration control (NH4 + with a setpoint of ∼8 mg N/L). Activity tests and FISH demonstrated that NOB barely survived in sludge flocs and were inhibited in biofilms. Despite receiving organic-deficient wastewater from a pilot-scale High-Rate Activated Sludge (HRAS) system as the feed, the system maintained a stable effluent total nitrogen concentration mostly below 10 mg N/L, which was attributed to the successful retention of anammox bacteria. This study successfully demonstrated large-scale long-term mainstream anammox application and generated new practical knowledge for NOB control and anammox retention.

Achieving robust mainstream nitrite shunt at pilot-scale with integrated sidestream sludge treatment and step-feed.

As a promising energy- and carbon efficient process for nitrogen removal from wastewater, mainstream nitrite shunt has been extensively researched. However, beyond the laboratory it is challenging to maintain stable performance by suppressing nitrite-oxidising bacteria (NOB). In this study, a pilot-scale reactor system receiving real sewage was operated in two stages for >850 days to evaluate two novel NOB suppression strategies for achieving nitrite shunt: i) sidestream sludge treatment based on alternating free nitrous acid (FNA) and free ammonia (FA) and ii) sidestream FNA/FA sludge treatment integrated with in-situ NOB suppression via step-feed. The results showed that, with sidestream sludge treatment alone, NOB developed resistance relatively quickly to the treatment, leading to unstable nitrite shunt. In contrast, robust nitrite shunt was achieved and stably maintained for more than a year when sidestream sludge treatment was integrated with a step-feed strategy. Kinetic analyses suggested that sludge treatment and step-feed worked in synergy, leading to stable NOB suppression. The integrated strategy demonstrated in this study removes a key barrier to the implementation of stable mainstream nitrite shunt.

Survival, sublethal injury, and recovery of environmental Burkholderia pseudomallei in soil subjected to desiccation.

Environmental Burkholderia pseudomallei isolated from sandy soil at Castle Hill, Townsville, in the dry tropic region of Queensland, Australia, was inoculated into sterile-soil laboratory microcosms subjected to variable soil moisture. Survival and sublethal injury of the B. pseudomallei strain were monitored by recovery using culture-based methods. Soil extraction buffer yielded higher recoveries as an extraction agent than sterile distilled water. B. pseudomallei was not recoverable when inoculated into desiccated soil but remained recoverable from moist soil subjected to 91 days' desiccation and showed a growth response to increased soil moisture over at least 113 days. Results indicate that endemic dry tropic soil may act as a reservoir during the dry season, with an increase in cell number and potential for mobilization from soil into water in the wet season.

A multicopper oxidase is essential for manganese oxidation and laccase-like activity in Pedomicrobium sp. ACM 3067.

Pedomicrobium sp. ACM 3067 is a budding-hyphal bacterium belonging to the alpha-Proteobacteria which is able to oxidize soluble Mn2+ to insoluble manganese oxide. A cosmid, from a whole-genome library, containing the putative genes responsible for manganese oxidation was identified and a primer-walking approach yielded 4350 bp of novel sequence. Analysis of this sequence showed the presence of a predicted three-gene operon, moxCBA. The moxA gene product showed homology to multicopper oxidases (MCOs) and contained the characteristic four copper-binding motifs (A, B, C and D) common to MCOs. An insertion mutation of moxA showed that this gene was essential for both manganese oxidation and laccase-like activity. The moxB gene product showed homology to a family of outer membrane proteins which are essential for Type I secretion in Gram-negative bacteria. moxBA has not been observed in other manganese-oxidizing bacteria but homologues were identified in the genomes of several bacteria including Sinorhizobium meliloti 1021 and Agrobacterium tumefaciens C58. These results suggest that moxBA and its homologues constitute a family of genes encoding an MCO and a predicted component of the Type I secretion system.