ABSTRACT
Simple cryopreservation of anaerobic ammonium-oxidation (anammox) consortia has become a promising preservation technology for the fast start-up of the anammox process. Here, we use genome-resolved metagenomics and metatranscriptomics to understand of the microbial interaction in a simple and effective resuscitation process for long-term cryopreserved anammox consortia by sequential addition of anammox SBR supernatant. Performance results showed that sequential addition of anammox supernatant significantly reduced the resuscitation time of the granule-based anammox process from 40 to 20 days. Genome-centric metagenomics were used to recover 19 high-quality draft genomes of anammox and heterotrophic bacteria. Comparative metatranscriptomic analysis was conducted to examine the gene expression of Candidatus Kuenenia stuttgartiensis, the dominant anammox bacterium, and heterotrophic bacteria to better understand their potential interactions. Proteobacteria-affiliated bacteria found in the supernatant were highly active in producing the secondary metabolites molybdopterin cofactor and folate which are needed for growth of the auxotrophic anammox bacteria. In addition, the significantly higher expression levels of hzsA and CO2-fixtion genes in the Candidatus Kuenenia genome indicated the anammox bacteria were likely more active and growing faster after sequential anammox supernatant addition during the resuscitation process. The resuscitation treatment pulse assays confirmed that sequential addition of supernatant was an effective way for the rapid resuscitation of anammox consortia. Our findings offer the first evidence of cross-feeding during the rapid resuscitation of cryopreserved anammox consortia.
Subject(s)
Bioreactors , Nitrogen , Anaerobiosis , Cryopreservation , Metagenomics , Oxidation-ReductionABSTRACT
This study was aimed to investigate the prevalence and genotype distribution of heterozygotes in beta-thalassemia combining deletional alpha-thalassemia by using molecular detection and haematological methods. Three common deletions of alpha-thalassemia were detected by using gap-PCR. The mutations of beta-thalassemia were identified by using PCR with reverse dot blot hybridization. The routine analysis of blood cells was carried out. The results indicated that 15 cases from the 81 beta-thalassemia traits were found to be the compound heterozygosity for beta-thalassemia and alpha-thalassemia with 9 different types of gene defects with 18.52% detection rate. There were 6 cases (7.41%) of beta-thalassemia heterozygote combining alpha-thalassemia-1 gene (--(SEA)/alphaalpha), 8 cases (9.88%) combining with alpha-thalassemia-2 gene including 6 (7.41%) right ward deletion (-alpha(3.7)/alphaalpha) and 2 (2.47%) left ward deletion (-alpha(4.2)/alphaalpha), and 1 case (1.23%) combining deletional HbH gene (--(SEA)/-alpha(3.7)). No significant differences were found between beta-thalassemia heterozygotes combining deletional alpha-thalassemia and pure beta-thalassemia in all RBC parameters. It is concluded that the incidence of beta-thalassemia heterozygotes combining with deletional alpha-thalassemia is frequent in Wuzhou city. The hematological analysis can not give specificity for diagnosing these dual heterozygotes. Gap-PCR as a routine method for thalassemia screening has the advantages in reducing the possibility of failing to detect the combining heterozygosity for beta-thalassemia and alpha-thalassemia. It is more useful for genetic counselling and prenatal diagnosis of this disease.
