Production and Excretion of Polyamines To Tolerate High Ammonia, a Case Study on Soil Ammonia-Oxidizing Archaeon "Candidatus Nitrosocosmicus agrestis".

Ammonia tolerance is a universal characteristic among the ammonia-oxidizing bacteria (AOB); in contrast, the known species of ammonia-oxidizing archaea (AOA) have been regarded as ammonia sensitive, until the identification of the genus "Candidatus Nitrosocosmicus." However, the mechanism of its ammonia tolerance has not been reported. In this study, the AOA species "Candidatus Nitrosocosmicus agrestis," obtained from agricultural soil, was determined to be able to tolerate high concentrations of NH3 (>1,500 µM). In the genome of this strain, which was recovered from metagenomic data, a full set of genes for the pathways of polysaccharide metabolism, urea hydrolysis, arginine synthesis, and polyamine synthesis was identified. Among them, the genes encoding cytoplasmic carbonic anhydrase (CA) and a potential polyamine transporter (drug/metabolite exporter [DME]) were found to be unique to the genus "Ca. Nitrosocosmicus." When "Ca. Nitrosocosmicus agrestis" was grown with high levels of ammonia, the genes that participate in CO2/HCO3 - conversion, glutamate/glutamine syntheses, arginine synthesis, polyamine synthesis, and polyamine excretion were significantly upregulated, and the polyamines, including putrescine and spermidine, had significant levels of production. Based on genome analysis, gene expression quantification, and polyamine determination, we propose that the production and excretion of polyamines is probably one of the reasons for the ammonia tolerance of "Ca. Nitrosocosmicus agrestis," and even of the genus "Ca. Nitrosocosmicus."IMPORTANCE Ammonia tolerance of AOA is usually much lower than that of the AOB, which makes the AOB rather than AOA a predominant ammonia oxidizer in agricultural soils, contributing to global N2O emission. Recently, some AOA species from the genus "Ca. Nitrosocosmicus" were also found to have high ammonia tolerance. However, the reported mechanism for the ammonia tolerance is very rare and indeterminate for AOB and for AOA species. In this study, an ammonia-tolerant AOA strain of the species "Ca. Nitrosocosmicus agrestis" was identified and its potential mechanisms for ammonia tolerance were explored. This study will be of benefit for determining more of the ecological role of AOA in agricultural soils or other environments.

The regulatory mechanism of Chryseobacterium sp. resistance mediated by montmorillonite upon cadmium stress.

Cadmium (Cd) is a toxic heavy metal and its uptake by living organisms causes adverse effect, further resulting in cycle pollution of the biosphere. The specific regulatory mechanism between clays and microbes under Cd stress remains unclear. In this study, interface interactions among clays, microbes and Cd were confirmed. Comparative transcriptome was conducted to investigate how it regulated gene expression patterns of microbes (Chryseobacterium sp. WAL2), which exposed to a series of gradient concentrations of Cd (16, 32, 64 and 128 µg mL-1) for 12 d in the presence and absence of clay montmorillonite (Mt) (16 g L-1). Cd was highly enriched by the unique interface interactions between Mt and bacteria (67.6-82.1%), leading to a more hostile environment for bacterial cells. However, Mt ultimately enhanced bacterial resistance to Cd stress by stimulating the mechanism of bacterial resistance; namely: (i) Mt increased genes expression connected with ion transport, enhancing the uptake of Cd; (ii) Mt stimulated genes expression related to efflux pump and positively regulated cellular oxidative stress (e.g., glutathione) and Cd accumulation (e.g., cysteine) processes. Further, genes expression related to intracellular metabolic processes was enforced, which supplied a driving force and accelerated electron transfer; (iii) Mt improved genes expression involved in DNA replication and other biological processes (e.g., terpenoid backbone biosynthesis) to maintain bacterial vitality. Therefore, the study not only optimized a unique Cd resistance mechanism of Mt on Chryseobacterium sp., but also provided a novel insight for environmental mitigation of heavy metals from the perspective of molecular biology.

A Two-Step Strategy for the Rapid Enrichment of Nitrosocosmicus-Like Ammonia-Oxidizing Thaumarchaea.

Ammonia-oxidizing archaea (AOA) are widely distributed on the earth and play a significant role in the global nitrogen cycle. Although dozens of AOA strains were obtained in the last 13 years, it is still necessary to obtain more AOA strains for the entire exploration of their ecology, physiology, and underlying biochemistry in different environments. In this study, we designed a two-step strategy for the rapid enrichment of Nitrosocosmicus-like AOA from soils. Firstly, combination of kanamycin and ampicillin was chosen as the selective stress for bacteria and quartz sands were used as the attachment of AOA cells during the first step cultivation; only after 40-75 days cultivation, AOA enrichments with abundance >20% were obtained. Secondly, combination of ciprofloxacin and azithromycin was chosen as the selective stress for the following cultivation; it is able to penetrate the biofilms and kill the bacterial cells inside the aggregate, contributing to the AOA enrichments reached high abundances (90%) only after one-time cultivation. Basing on this strategy, three AOA strains were obtained from agricultural soils only after 90-150 days cultivation. Phylogenetic analysis suggested these AOA belong to the soil group I.1b Thaumarchaeota and are closely related to the genus Nitrosocosmicus. In general, AOA enrichment or isolation is very difficult and time-consuming (an average of 2-3 years). Here, we provide a new strategy for the rapid enrichment of high abundance of Nitrosocosmicus-like AOA from soil, which gives a new solution to the AOA enrichment and cultivation in a short period.

Insight into the diversity of antibiotic resistance genes in the intestinal bacteria of shrimp Penaeus vannamei by culture-dependent and independent approaches.

Antibiotic resistance genes (ARGs) that distributed in antibiotic resistant bacteria (ARBs) are widespread in aquaculture and have great threats to the aquatic organism as well as to human. However, our understanding about the risk of ARGs to the health of aquatic organism is still limited. In the present study, we got a deep insight into the diversity of ARGs in the intestinal bacteria of shrimp by culture-dependent and independent approaches. Results of the PCR-based detection and culture-dependent analysis indicated that the tetracycline, sulfadiazine, quinolone and erythromycin resistance genes were prevalent in the commercial shrimps that bought from aquatic markets or supermarket. The culture-independent plasmid metagenomic analysis identified 62 different ARGs, which were classified into 21 types, with abundances ranging from 13 to 1418 ppm. The analysis suggested that most of the ARGs come from the plasmids originating from Vibrio (accounted for 2.8-51%) and Aeromonas (accounted for 16-55%), and the Vibrio group was concluded to be the main bacterial pathogen that probably resulted in the shrimp disease. Accordingly, the plasmid metagenomic that focuses on the mobile genetic elements has great potential on the identification of ARGs in complex environments.

Potential of using sodium bicarbonate as external carbon source to cultivate microalga in non-sterile condition.

In this study, a saline-alkaline tolerant microalgal strain was isolated and identified as Chlorella sp. LPF. This strain was able to grow at pH values up to 10 and at salinities up to 5%, and tolerated to 80 g L-1 of sodium bicarbonate. The utilization of bicarbonate as carbon source significantly promoted microalgal growth and lipid production. In the non-sterile cultivation supplying with 80 g L-1 of sodium bicarbonate, the microalgal growth had no difference with their growth in the sterile medium; however, the bacterial growth was suppressed and the cell number decreased to low levels after six days cultivation. This study gives an insight into the potential that using high concentration of sodium bicarbonate as external carbon source to cultivate microalga in non-sterile condition, and suggests a possibility of using bicarbonate as growth promoter and antibacterial agent for the microalgal outdoor cultivation.