Acinetobacter baumannii is an opportunistic pathogen often found in patients with low immunity. It causes nosocomial infections, which are difficult to treat. This bacterium can rapidly mutate, developing resistance to antimicrobials and adapting to environmental stress, thereby increasing its survival. Understanding such adaptive mechanisms will be beneficial for controlling the spread of A. baumannii. Astrobiology studies have demonstrated that microbiomes from astronauts and manned spaceflight environments show resistance to stress and antibiotics. Astronauts also encounter low immunity during spaceflight missions. The extreme conditions of spaceflight provide a unique research platform for studying how opportunistic pathogens such as A. baumannii adapt to conditions such as microgravity and mutate during spaceflight. In this study, we compared phenotypic variations and analyzed genomic and transcriptomic variations in A. baumannii strains exposed to three different conditions: ST1 (64 days on Tiangong-2 space laboratory), GT1 (ground control), and Aba (original strain). Biofilm formation ability of the ST1 strain increased after 64 days of spaceflight. In addition, high-throughput sequencing revealed that some differentially expressed genes were upregulated in the ST1 strain compared to the GT1 strain. These results provide insights into the environmental adaptation of this widespread pathogen.

AIM:This study aimed to explore potential tobramycin-resistant mutagenesis of Escherichia coli strains after spaceflight.MATERIALS & METHODS:A spaceflight-induced mutagenesis of multidrug resistant E. coli strain (T1_13) on the outer space for 64 days (ST5), and a ground laboratory with the same conditions (GT5) were conducted. Both whole-genome sequencing and RNA-sequencing were performed.RESULTS:A total of 75 single nucleotide polymorphisms and 20 InDels were found to be associated with the resistance mechanism. Compared with T1_13, 1242 genes were differentially expressed in more than 20 of 38 tobramycin-resistant E. coli isolates while not in GT5. Function annotation of these single nucleotide polymorphisms/InDels related genes and differentially expressed genes was performed.CONCLUSION:This study provided clues for potential tobramycin-resistant spaceflight-induced mutagenesis of E. coli.

China launched the Tiangong-2 space laboratory in 2016 and will eventually build a basic space station by the early 2020s. These spaceflight missions require astronauts to stay on the space station for more than 6 months, and they inevitably carry microbes into the space environment. It is known that the space environment affects microbial behavior, including growth rate, biofilm formation, virulence, drug resistance, and metabolism. However, the mechanisms of these alternations have not been fully elucidated. Therefore, it is beneficial to monitor microorganisms for preventing infections among astronauts in a space environment. Salmonella enteritidis is a Gram-negative bacterial pathogen that commonly causes acute gastroenteritis in humans. In this study, to better understand the effects of the space environment on S. enteritidis, a S. enteritidis strain was taken into space by the Shenzhou-11 spacecraft from 17 October 2016 to 18 November 2016, and a ground simulation with similar temperature conditions was simultaneously performed as a control. It was found that the flight strain displayed an increased growth rate, enhanced amikacin resistance, and some metabolism alterations compared with the ground strain. Enrichment analysis of proteome revealed that the increased growth rate might be associated with differentially expressed proteins involved in transmembrane transport and energy production and conversion assembly. A combined transcriptome and proteome analysis showed that the amikacin resistance was due to the downregulation of the oppA gene and oligopeptide transporter protein OppA. In conclusion, this study is the first systematic analysis of the phenotypic, genomic, transcriptomic, and proteomic variations in S. enteritidis during spaceflight and will provide beneficial insights for future studies on space microbiology.

BACKGROUND:Multidrug resistance is a growing global public health threat with far more serious consequences than generally anticipated. In this study, we investigated the antibiotic resistance and genomic traits of a clinical strain of Escherichia coli LCT-EC001. RESULTS:LCT-EC001 was resistant to 16 kinds of widely used antibiotics, including fourth-generation cephalosporins and carbapenems. In total, up to 68 determinants associated with antibiotic resistance were identified, including 8 beta-lactamase genes (notably producing ESBLs and KPCs), 31 multidrug efflux system genes, 6 outer membrane transport system genes, 4 aminoglycoside-modifying enzyme genes, 10 two-component regulatory system genes, and 9 other enzyme or transcriptional regulator genes, covering nearly all known drug-resistance mechanisms in E. coli. More than half of the resistance genes were located close to mobile genetic elements, such as plasmids, transposons, genomics islands, and insertion sequences. Phylogenetic analysis revealed that this strain may have evolved from E. coli K-12 but is a completely new MLST type. CONCLUSIONS:Antibiotic resistance was extremely severe in E. coli LCT-EC001, mainly due to mobile genetic elements that allowed the gain of a large quantity of resistance genes. The antibiotic resistance genes of E. coli LCT-EC001 can probably be transferred to other bacteria. To the best of our knowledge, this is the first report of a strain of E. coli which has such a large amount of antibiotic resistance genes. Apart from providing an E. coli reference genome with an extremely high multidrug-resistant background for future analyses, this work also offers a strategy for investigating the complement and characteristics of genes contributing to drug resistance at the whole-genome level.

Background Many studies have shown that space environment can affect bacteria to cause a range of mutations. However, so far, there are less studies on the effects of long-term space flight (>1 month) on bacteria. In this study, a Staphylococcus warneri strain, which was isolated from the Shenzhou-10 spacecraft that had experienced a space flight (15-day), was carried into space again. After 64-day flight, the combined phenotypic, genomic, transcriptomic and proteomic analyses were performed to compare the influence of the two space flights on this bacteria.Results Compared with short-term space flight, long-term space flight increased the biofilm formation ability of S. warneri and the cell wall resistance to external environment stress, but reduced the sensitivity to chemical stimulation. Further analysis showed that these changes might be related to the significantly up-regulated gene expression of phosphotransferase system, which regulated glucose metabolism pathway, including glucose, mannose, fructose and cellobiose. The mutation of S. warneri caused by 15-day space flight was limited at phenotype and gene level after ground culture.Conclusion After a 79-day space flight, the changes of S. warneri are meaningful. Phosphotransferase system of S. warneri was up-regulated by long-term space stimulation, which resulted in a series of changes in cell wall, biofilm and chemical sensitivity, thus enhancing the resistance and adaptability of bacteria to the external environment.Importance Taking S. warneri as an example, we found that the mutation of bacteria in long-term space flight mainly focused on the adaptation and tolerance like environmental adaptation, physical resistance and cell repair to various pressures and these changes should be more obvious or even become stable mutations. But the clinical changes, for example drug resistance and virulence should not be significant because there is no threat of antibiotics.

BACKGROUND: Space is a special environment in which microgravity and cosmic rays are the primary factors that induce gene mutations of microorganisms. In our previous studies, a single point mutation in the gene dprA was found in an Enterococcus faecium strain of LCT-EF258 after spaceflight. DNA processing protein A (DprA) plays a prominent role in the horizontal transfer of genes among bacteria (such as Streptococcus pneumoniae, Helicobacter pylori, Bacillus subtilis, and Rhodobacter capsulatus). However, the function of DprA in E. faecium remains unknown. Furthermore, E. faecium could acquire antibiotic resistance through the horizontal transfer of antibiotic resistance genes, but it is unclear whether dprA mutants could affect this process in E. faecium.METHODS: In this study, we constructed a plasmid containing the vancomycin resistance gene vanA and then transferred the gene vanA into the dprA-mutant strain LCT-EF258 and the control strain LCT-EF90 using the electroporation technique. We then used Discovery StudioTM software to construct the 3D protein structure.RESULTS: The results showed that the horizontal transfer efficiency of the vancomycin resistance gene vanA in the dprA-mutant E. faecium decreased. And the hydrophobic core of the mutant DprA became stable and the binding affinity between the mutant DprA and ssDNA reduced.DISCUSSION: This study is an exploration of bacterial gene mutation after spaceflight. The dprA mutant could affect the ability of E. faecium to acquire exogenous resistance gene vanA, which offered us an interesting path to block the dissemination of resistance genes between strains.Yu Y, Chang D, Guo Q, Wang J, Liu C. LCT-EF258 with S171 mutation in DprA exhibits horizontal gene transfer deficiency after spaceflight. Aerosp Med Hum Perform. 2019; 90(2):116-122.

Background Acetylsalicylic acid was renamed aspirin in 1899, and it has been widely used for its multiple biological actions. Because of the diversity of the cellular processes and diseases that aspirin reportedly affects and benefits, uncertainty remains regarding its mechanism in different biological systems. Methods The Drugbank and STITCH databases were used to find direct protein targets (DPTs) of aspirin. The Mentha database was used to analyze protein–protein interactions (PPIs) to find DPT-associated genes. DAVID was used for the GO and KEGG enrichment analyses. The cBio Cancer Genomics Portal database was used to mine genetic alterations and networks of aspirin-associated genes in cancer. Results Eighteen direct protein targets (DPT) and 961 DPT-associated genes were identified for aspirin. This enrichment analysis resulted in eight identified KEGG pathways that were associated with cancers. Analysis using the cBio portal indicated that aspirin might have effects on multiple tumor suppressors, such as TP53, PTEN, and RB1 and that TP53 might play a central role in aspirin-associated genes. Discussion The results not only suggest that aspirin might have anti-tumor actions against multiple cancers but could also provide new directions for further research on aspirin using a bioinformatics analysis approach.

As a highly drug-resistant and widely spread pathogen, carbapenemase (KPC)-producing Klebsiella pneumoniae represents an urgent public health crisis. Variations in the ability to cope with environmental hazards increase a microbe's survival chances. Therefore, understanding such adaptive mechanisms will be beneficial for controlling the spread K. pneumoniae and the development of anti-infective therapies. The extreme environment in spaceflight represents a unique research platform. We compared phenotypic variations and analyzed genomic and transcriptomic variations in KPC-2 K. pneumoniae among three different groups: strains T (15 days of Shenzhou X spaceflight), D (the ground control) and F (the reference strain). We found no obvious variations in the drug susceptibility, carbon source utilization, biofilm-forming capacity, and shapes and arrangements of bacterial cells of the D and T strains; however, the growth rates and final concentrations of the bacterial cells decreased when cultured in hypertonic LB medium (8% sodium chloride). Additionally, high-throughput sequencing revealed some genomic mutations and differentially regulated transcriptomes between the D and T strains. These results provide insights into the environmental adaptation of this widespread pathogen.

AbstractChina has prepared for construction of a space station by the early 2020s. The mission will require astronauts to stay on the space station for at least 180 days. Microbes isolated from the International Space Station (ISS) have shown profound resistance to clinical antibiotics and environmental stresses. Previous studies have demonstrated that the space environment could affect microbial survival, growth, virulence, biofilms, metabolism, as well as their antibiotic‐resistant phenotypes. Furthermore, several studies have reported that astronauts experience a decline in their immunity during long‐duration spaceflights. Monitoring microbiomes in the ISS or the spacecraft will be beneficial for the prevention of infection among the astronauts during spaceflight. The development of a manned space program worldwide not only provides an opportunity to investigate the impact of this extreme environment on opportunistic pathogenic microbes, but also offers a unique platform to detect mutations in pathogenic bacteria. Various microorganisms have been carried on a spacecraft for academic purposes. Acinetobacter baumannii is a common multidrug‐resistant bacterium often prevalent in hospitals. Variations in the ability to cope with environmental hazards increase the chances of microbial survival. Our study aimed to compare phenotypic variations and analyze genomic and transcriptomic variations in A. baumannii among three different groups: SS1 (33 days on the Shenzhou 11 spacecraft), GS1 (ground control), and Aba (reference strain). Consequently, the biofilm formation ability of the SS1 strain decreased after 33 days of spaceflight. Furthermore, high‐throughput sequencing revealed that some differentially expressed genes were downregulated in the SS1 strain compared with those in the GS1 strain. In conclusion, this present study provides insights into the environmental adaptation of A. baumannii and might be useful for understanding changes in the opportunistic pathogenic microbes on our spacecraft and on China's future ISS.

Streptococcus pneumoniae is one of the most frequent opportunistic pathogens worldwide. DNA processing protein A (DprA) is an important factor involved in bacterial uptake and DNA integration into bacterial genome, but its role in S. pneumoniae virulence remains unclear. The aim of this study was to characterize the effects of the pneumococcal dprA gene on the pathogenesis of S. pneumoniae. To construct a dprA-deficient pneumococcal strain, the dprA gene of the S. pneumoniae strain D39 was inactivated. The virulence of this dprA-deficient strain, designated ΔD39, was compared with that of the wild-type strain by evaluating their respective capabilities to adhere to human pulmonary epithelial cells (PEC-A549) and by analyzing their choline-binding protein expression levels. In addition, the expression profiles of genes associated with virulence and host survival assays were also conducted with the mutant and the wild-type strain. Our results indicate that the capability of ΔD39 to adhere to the PEC-A549 airway cells was significantly lower (p<0.01) compared with D39. Additionally, the 100-KD choline-binding protein was not detected in ΔD39. The addition of competence-stimulating peptide (CSP) lead to a significantly reduction of psaA mRNA expression in the dprA-deficient mutant and an increased level of psaA transcripts in the wild-type strain (p<0.01). The median survival time of mice intraperitoneally infected with ΔD39 was significantly higher (p<0.01) than that of mice infected with D39. The results of this study suggest that DprA has a significant effect on virulence characteristics of S. pneumoniae by influencing the expression of choline-binding protein and PsaA.

Endoproteinase Lys-C is an important tool enzyme in industrial produce, mainly gained from Lysobacterenzymogenesat a low efficiency and a long fermentation period. The special environmental factors of outer space may stimulate engineering bacteria to bear high-yield mutant strains. In this study, we sent Lenzymogenes into space via the Chinese Shenzhou-10 spacecraft for approximate 15 days,finding that the total cell number reduced to (24 +/- 3.5) x10(5)cfu/ml after spaceflight, only 13.5% of that in the ground control sample. However, Lys-C activityin ground control bacteria was 0.087 U/ml after fermentation for 120 hours, which was its optical fermentation time for maximal yield, while that in spaceflight sample increased by 17.2%-40.2%,with theoptical fermentation time shortened by 24-48 hours. Further investigating the genetic stability of spaceflight sample revealed that both enzyme-producing ability and optical fermentation time after 1, 3, 5, 7, 9, 11 and 15 passages didn't change,revealing good stability of the spaceflight mutant strain. In summary, spaceflight experience could stably increase the yield of LysC by L.enzymogenes and shorten the optical fermentation time of Lys-C, which will benefit its industrial produce.

Six strains of bacteria, comprising three Gram-negative and three Gram-positive strains, were utilised to reveal the functional relationship between optical density (OD) values and colony-forming units (CFU) during the exponential growth phase. CFU during the exponential growth phase were counted using the classic plate counting method. OD values were detected with both Multiskan Spectrum and N721 spectrophotometers at 600 nm at both 37 degrees C and 4 degrees C. Two functions (line function and power function) were simulated with Excel 2007 to reveal the relationship between OD values and the CFU. The two functions were compared with each other by using their coefficient of determination values (R-2), which represented degree of fitting for functions. For the ODs detected with the Multiskan Spectrum spectrophotometer at 37 degrees C, the average R-2 values of the line function was 0.983+/-0.011 and the power function was 0.995+/-0.003; thus, the R-2 value of the power function was significantly higher than that of the line relationship (p<0.05). Similar results were obtained with the Multiskan Spectrum spectrophotometer at 4 degrees C and with the N721 spectrophotometer at both temperatures. These results reveal that the power function is more suitable for modelling the relationship between OD600 and CFU during the exponential growth phase of bacteria.

ABSTRACT Escherichia coli is a ubiquitous opportunistic pathogen that colonizes the lower intestines of humans and causes several diseases, such as septicemia, pneumonia, and urinary tract infections. Here, we present the draft genome sequence of E. coli strain LCT-EC52, which originated from E. coli strain CGMCC 1.2385 and acquired changes in antibiotic resistance following travel on the Shenzhou-VIII spacecraft.