Emergence of aztreonam/avibactam and tigecycline-resistant Pseudomonas putida group Co-producing blaIMP-1, blaAFM-4 and blaOXA-1041 with a novel sequence type ST268 in Southwestern China.

OBJECTIVES: The emergence of carbapenem-resistant Pseudomonas putida (CRPP) has raised public awareness. This study investigated two strains from the Pseudomonas putida group that were resistant to carbapenem, tigecycline, and aztreonam-avibactam (ATM-AVI), with a focus on their microbial and genomic characteristics. METHODS: We assessed the antibiotic resistance profile using broth dilution, disk diffusion, and E-test methods. Efflux pump phenotype testing and real-time quantitative PCR were employed to evaluate efflux pump activity in tigecycline resistance, while polymerase chain reaction was utilized to detect common carbapenem genes. Additionally, whole-genome sequencing was performed to analyze genomic characteristics. The transferability of blaIMP-1 and blaAFM-4 was assessed through a conjugation experiment. Furthermore, growth kinetics and biofilm formation were examined using growth curves and crystal violet staining. RESULTS: Both strains demonstrated resistance to carbapenem, tigecycline, and ATM-AVI. Notably, NMP can restore sensitivity to tigecycline. Subsequent analysis revealed that they co-produced blaIMP-1, blaAFM-4, tmexCD-toprJ, and blaOXA-1041, belonging to a novel sequence type ST268. Although they were closely related on the phylogenetic tree, they exhibited different levels of virulence. Genetic environment analysis indicated variations compared to prior studies, particularly regarding the blaIMP-1 and blaAFM-4 genes, which showed limited horizontal transferability. Moreover, it was observed that temperature exerted a specific influence on their biological factors. CONCLUSION: We initially identified two P. putida ST268 strains co-producing blaIMP-1, blaAFM-4, blaOXA-1041, and tmexCD-toprJ. The resistance to tigecycline and ATM-AVI can be attributed to the presence of multiple drug resistance determinants. These findings underscore the significance of P. putida as a reservoir for novel antibiotic resistance genes. Therefore, it is imperative to develop alternative antibiotic therapies and establish effective monitoring of bacterial resistance.

Role of efflux pumps, their inhibitors, and regulators in colistin resistance.

Colistin is highly promising against multidrug-resistant and extensively drug-resistant bacteria clinically. Bacteria are resistant to colistin mainly through mcr and chromosome-mediated lipopolysaccharide (LPS) synthesis-related locus variation. However, the current understanding cannot fully explain the resistance mechanism in mcr-negative colistin-resistant strains. Significantly, the contribution of efflux pumps to colistin resistance remains to be clarified. This review aims to discuss the contribution of efflux pumps and their related transcriptional regulators to colistin resistance in various bacteria and the reversal effect of efflux pump inhibitors on colistin resistance. Previous studies suggested a complex regulatory relationship between the efflux pumps and their transcriptional regulators and LPS synthesis, transport, and modification. Carbonyl cyanide 3-chlorophenylhydrazone (CCCP), 1-(1-naphthylmethyl)-piperazine (NMP), and Phe-Arg-ß-naphthylamide (PAßN) all achieved the reversal of colistin resistance, highlighting the role of efflux pumps in colistin resistance and their potential for adjuvant development. The contribution of the efflux pumps to colistin resistance might also be related to specific genetic backgrounds. They can participate in colistin tolerance and heterogeneous resistance to affect the treatment efficacy of colistin. These findings help understand the development of resistance in mcr-negative colistin-resistant strains.

Species differences in stoichiometric homeostasis affect grassland community stability under N and P addition.

Unbalanced N and P input has substantially altered the relative importance of N and P limitation in grassland ecosystems, which resulted in profound impacts on species nutrient cycling, community structure, and ecosystem stability. However, the underlying species-specific nutrient use strategy and stoichiometric homeostasis in driving community structure and stability changes remain unclear. A split-plot N and P addition experiment (main-plot: 0, 25, 50, and 100 kgN hm-2 a-1; subplot: 0, 20, 40, and 80 kgP2O5 hm-2 a-1) was conducted during 2017-2019 in two typical grasslands (perennial grass and perennial forb) communities in the Loess Plateau. The stoichiometric homeostasis of 10 main component species, species dominance, stability changes, and their contribution to community stability were investigated. Perennial legume and perennial clonal species tend to perform higher stoichiometric homeostasis than non-clonal and annual forb. Large shifts in species with high homeostasis vs. low homeostasis caused by N and P addition showed consistently profound impacts on community homeostasis and stability in both communities. In both two communities, species dominance performed significantly positive relationships with homeostasis under no N and P addition. P alone or combined with 25 kgN hm-2 a-1 addition strengthened species dominance-homeostasis relationship and increased community homeostasis due to increased perennial legumes. Under 50 and 100 kgN hm-2 a-1 combined with P addition, species dominance-homeostasis relationships were weakened, and community homeostasis decreased significantly in both communities, which was due to that increased annual and non-clonal forb suppressed perennial legume and clonal species. Our results demonstrated that trait-based classifications of species-level homeostasis offer a reliable tool in predicting species performance and community stability under N and P addition, and conserving species with high homeostasis is important to enhance semiarid grassland ecosystem function stability on the Loess Plateau.

Carbapenem-resistant Acinetobacter baumannii: A challenge in the intensive care unit.

Carbapenem-resistant Acinetobacter baumannii (CRAB) has become one of the leading causes of healthcare-associated infections globally, particularly in intensive care units (ICUs). Cross-transmission of microorganisms between patients and the hospital environment may play a crucial role in ICU-acquired CRAB colonization and infection. The control and treatment of CRAB infection in ICUs have been recognized as a global challenge because of its multiple-drug resistance. The main concern is that CRAB infections can be disastrous for ICU patients if currently existing limited therapeutic alternatives fail in the future. Therefore, the colonization, infection, transmission, and resistance mechanisms of CRAB in ICUs need to be systematically studied. To provide a basis for prevention and control countermeasures for CRAB infection in ICUs, we present an overview of research on CRAB in ICUs, summarize clinical infections and environmental reservoirs, discuss the drug resistance mechanism and homology of CRAB in ICUs, and evaluate contemporary treatment and control strategies.

Leaf Photosynthetic and Functional Traits of Grassland Dominant Species in Response to Nutrient Addition on the Chinese Loess Plateau.

Leaf photosynthetic and functional traits of dominant species are important for understanding grassland community dynamics under imbalanced nitrogen (N) and phosphorus (P) inputs. Here, the effects of N (N0, N50, and N100, corresponding to 0, 50, and 100 kg ha-1 yr-1, respectively) or/and P additions (P0, P40, and P80, corresponding to 0, 40, and 80 kg ha-1 yr-1) on photosynthetic characteristics and leaf economic traits of three dominant species (two grasses: Bothriochloa ischaemum and Stipa bungeana; a leguminous subshrub: Lespedeza davurica) were investigated in a semiarid grassland community on the Loess Plateau of China. Results showed that, after a three-year N addition, all three species had higher specific leaf area (SLA), leaf chlorophyll content (SPAD value), maximum net photosynthetic rate (PNmax), and leaf instantaneous water use efficiency (WUE), while also having a lower leaf dry matter content (LDMC). The two grasses, B. ischaemum and S. bungeana, showed greater increases in PNmax and SLA than the subshrub L. davurica. P addition alone had no noticeable effect on the PNmax of the two grasses while it significantly increased the PNmax of L. davurica. There was an evident synergetic effect of the addition of N and P combined on photosynthetic traits and most leaf economic traits in the three species. All species had relatively high PNmax and SLA under the addition of N50 combined with P40. Overall, this study suggests that N and P addition shifted leaf economic traits towards a greater light harvesting ability and, thus, elevated photosynthesis in the three dominant species of a semiarid grassland community, and this was achieved by species-specific responses in leaf functional traits. These results may provide insights into grassland restoration and the assessment of community development in the context of atmospheric N deposition and intensive agricultural fertilization.

Effects of plant diversity on semiarid grassland stability depends on functional group composition and dynamics under N and P addition.

Exogenous fertilization could efficiently improve grassland productivity and promote grassland restoration. Increasing fertilization may profoundly affect community stability, whereas the underlying compensatory dynamics among functional groups in regulating grassland stability remain unclear. Three different grasslands, annuals forb (AF) community, perennial grass (PG) community and perennial forb (PF) community, on semiarid Loess Plateau were selected. We designed a 3-year split-plot experiment (main-plot: 0, 25, 50, and 100 kg N ha-1 yr-1; subplot: 0, 20, 40 and 80 kg P2O5 ha-1 yr-1) to explore how N and P addition affects community stability and its relationship with species richness, species asynchrony and functional group stability. Temporal stability differed largely between functional groups under N and P addition, perennial forbs or grasses had higher stability than perennial legumes or annuals and biennials. Decreased stability of PG and PF communities was primarily due to reduced species asynchrony under N addition alone, while it attributed to increased dominance of perennial legumes after P addition alone. 50 and 100 kg N ha-1 yr-1 combined with P addition significantly increased dominance of annuals and biennials, but decreased stability of annuals and biennials, which caused significant declines in stability of the three communities. Significant species richness decline induced by N and P addition only occurred in AF community, which suppressed AF community stability through reducing species asynchrony. AF community stability was regulated by additively negative effect of diversity decline and decreased annuals and biennials stability. Whereas, in PG and PF communities, nutrient-induced changes of functional groups stability were the main driver of community stability rather than diversity. Our study highlights the role of functional group composition and dynamics in regulating the effects of diversity on community stability and rational N and P combined addition was essential for conserving stability of different grasslands on semiarid Loess Plateau.

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To analyze plant functional traits of dominant species to nitrogen and phosphorus addition, three species (Bothriochloa ischaemum, Stipa bungeana, and Lespedeza davurica) were selected in the loess hilly-gully region. A split-plot experiment which included three N treatments (0, 50, and 100 kg N·hm-2·a-1) and three P treatments (0, 40, and 80 kg P2O5·hm-2·a-1) was conducted. At the fast-growing stage, leaf length, leaf width, specific leaf area, leaf dry matter content, leaf N content, leaf P content, and leaf N:P were measured. Results showed that under 50 and 100 kg N·hm-2·a-1 treatments, leaf length and width of B. ischaemum increased significantly by 35.3% and 64.4%, respectively, while only the leaf length of S. bungeana and the leaf width of L. davurica increased significantly by 58.8% and 33.9%, respectively. Leaf dry matter content of the three species decreased significantly by 10.7%, 15.3% and 11.2%, respectively. Leaf N content and N:P of B. ischaemum and S. bungeana increased significantly by 23.0% and 99.2%, 45.8% and 96.9%, respectively, compared with unfertilized treatments. Under 40 and 80 kg P2O5·hm-2·a-1 treatments, leaf length, leaf width and specific leaf area of L. davurica increased significantly by 56.9%, 41.4% and 19.6%, respectively, while leaf dry matter content decreased significantly by 14.9%. Leaf P content of three species increased significantly by 96.7%, 110.9% and 238.4%, while the N:P decreased significantly by 45.8%, 42.8% and 53.7%, respectively, compared with those under unfertilized. Under 50 kg N·hm-2·a-1 treatment, compared with no P application, leaf length and leaf width of L. davurica and leaf P content of the three species significantly increased, and leaf N content of B. ischaemum and S. bungeana decreased significantly at 40 and 80 kg P2O5·hm-2·a-1 treatments. Under 100 kg N·hm-2·a-1 treatment, leaf length of B. ischaemum and S. bungeana, leaf width of L. davurica and leaf P content of three species significantly increased, while leaf N content of B. ischaemum decreased significantly after P application. In summary, functional traits of dominant species showed significant responses to short-term nitrogen and phosphorus addition, with the different responses were mainly related to species traits and fertilization levels. Such difference reflected plant adaptation to habitat changes. The divergent responses of different species to nitrogen and phosphorus addition played an important role in maintaining diversity and stability of grassland communities.