Genetic Foundations of Direct Ammonia Oxidation (Dirammox) to N2 and MocR-Like Transcriptional Regulator DnfR in Alcaligenes faecalis Strain JQ135.

Ammonia oxidation is an important process in both the natural nitrogen cycle and nitrogen removal from engineered ecosystems. Recently, a new ammonia oxidation pathway termed Dirammox (direct ammonia oxidation, NH3→NH2OH→N2) has been identified in Alcaligenes ammonioxydans. However, whether Dirammox is present in other microbes, as well as its genetic regulation, remains unknown. In this study, it was found that the metabolically versatile bacterium Alcaligenes faecalis strain JQ135 could efficiently convert ammonia into N2 via NH2OH under aerobic conditions. Genetic deletion and complementation results suggest that dnfABC is responsible for the ammonia oxidation to N2 in this strain. Strain JQ135 also employs aerobic denitrification, mainly producing N2O and trace amounts of N2, with nitrite as the sole nitrogen source. Deletion of the nirK and nosZ genes, which are essential for denitrification, did not impair the capability of JQ135 to oxidize ammonia to N2 (i.e., Dirammox is independent of denitrification). Furthermore, it was also demonstrated that pod (which encodes pyruvic oxime dioxygenase) was not involved in Dirammox and that AFA_16745 (which was previously annotated as ammonia monooxygenase and is widespread in heterotrophic bacteria) was not an ammonia monooxygenase. The MocR-family transcriptional regulator DnfR was characterized as an activator of the dnfABC operon with the binding motif 5'-TGGTCTGT-3' in the promoter region. A bioinformatic survey showed that homologs of dnf genes are widely distributed in heterotrophic bacteria. In conclusion, this work demonstrates that, besides A. ammonioxydans, Dirammox occurs in other bacteria and is regulated by the MocR-family transcriptional regulator DnfR. IMPORTANCE Microbial ammonia oxidation is a key and rate-limiting step of the nitrogen cycle. Three previously known ammonia oxidation pathways (i.e., nitrification, anaerobic ammonia oxidation [Anammox], and complete ammonia oxidation [Comammox]) are mediated by autotrophic microbes. However, the genetic foundations of ammonia oxidation by heterotrophic microorganisms have not been investigated in depth. Recently, a previously unknown pathway, termed direct ammonia oxidation to N2 (Dirammox), has been identified in the heterotrophic bacterium Alcaligenes ammonioxydans HO-1. This paper shows that, in the metabolically versatile bacterium Alcaligenes faecalis JQ135, the Dirammox pathway is mediated by dnf genes, which are independent of the denitrification pathway. A bioinformatic survey suggests that homologs of dnf genes are widely distributed in bacteria. These findings enhance the understanding of the molecular mechanisms of heterotrophic ammonia oxidation to N2.

Efficacy of long-term orthokeratology treatment in children with anisometropic myopia.

AIM: To explore the efficacy of the orthokeratology lens for anisometropic myopia progression. METHODS: A retrospective study was performed. Cycloplegic refraction and axial length (AL) were collected from 50 children (10.52±1.72y) who visited Peking University Third Hospital from July 2015 to August 2020. These children's one eyes (Group A) received monocular orthokeratology lenses at first, after different durations (12.20±6.94mo), their contralateral eyes (Group B) developed myopia and receive orthokeratology as well. The data in 1-year of binocular period were recorded. AL growth rate (difference of follow-up and baseline per month) were compared between two groups by paired t test. Interocular differences of AL were compared by Wilcoxon test. RESULTS: During monocular period, the AL growth rate of the Group A (0.008±0.022 mm/mo) was significantly slower than that of the Group B (0.038±0.018 mm/mo; P<0.0001). However, during binocular period, the AL growth rate of the Group A (0.026±0.014 mm/mo) was significantly faster than that of the Group B (0.016±0.015 mm/mo; P<0.0001). The AL difference between both eyes was 0.6 (0.46) mm, then significantly decreased to 0.22 (0.39) mm when started binocular treatment (P<0.0001). However, it was significantly increased to 0.30 (0.32) mm after a year (P<0.0001), but still significantly lower than baseline (P<0.0001). CONCLUSION: The orthokeratology lens is efficient for control the AL elongation of monocular myopia eyes and reduce anisometropia. For the condition that the contralateral eyes develop myopia and receive orthokeratology lens later, there is no efficiency observed on control interocular difference of AL during binocular treatment.