The Combination of Antibiotic and Non-Antibiotic Compounds Improves Antibiotic Efficacy against Multidrug-Resistant Bacteria.

Bacterial antibiotic resistance, especially the emergence of multidrug-resistant (MDR) strains, urgently requires the development of effective treatment strategies. It is always of interest to delve into the mechanisms of resistance to current antibiotics and target them to promote the efficacy of existing antibiotics. In recent years, non-antibiotic compounds have played an important auxiliary role in improving the efficacy of antibiotics and promoting the treatment of drug-resistant bacteria. The combination of non-antibiotic compounds with antibiotics is considered a promising strategy against MDR bacteria. In this review, we first briefly summarize the main resistance mechanisms of current antibiotics. In addition, we propose several strategies to enhance antibiotic action based on resistance mechanisms. Then, the research progress of non-antibiotic compounds that can promote antibiotic-resistant bacteria through different mechanisms in recent years is also summarized. Finally, the development prospects and challenges of these non-antibiotic compounds in combination with antibiotics are discussed.

Diagnosis of Alzheimer's disease by feature weighted-LSTM: a preliminary study of temporal features in brain resting-state fMRI.

The long short-term memory network (LSTM) is widely used in time series data processing as a temporal recursive network. The resting-state functional magnetic resonance data shows that not only are there temporal variations in the resting state, but there are also interactions between brain regions. To integrate the temporal and spatial characteristics of brain regions, this paper proposes a model called feature weighted-LSTM (FW-LSTM). The feature weight is defined by spatial characteristics calculating the frequency of connectivity of each brain region and further integrated into the LSTM. Thus, it can comprehensively model both temporal and spatial changes in rs-fMRI brain regions. The FW-LSTM model on the Alzheimer's disease neuroimaging initiative (ADNI) dataset is used to extract the time-varying characteristics of 90 brain regions for Alzheimer's disease (AD) classification. The model performances are 77.80%, 76.41%, and 78.81% in accuracy, sensitivity, and specificity. It outperformed the one-dimensional convolutional neural networks (1D-CNN) model and LSTM model, which only used temporal features of brain regions.

Emergence of livestock-associated MRSA ST398 from bulk tank milk, China.

OBJECTIVES: To detect livestock-associated MRSA (LA-MRSA) ST398 from bulk tank milk in China and to determine the phenotypic and genomic characteristics of the strains. METHODS: LA-MRSA ST398 strains were isolated from bulk tank milk samples in Shanghai and their susceptibilities to antimicrobials were determined using the broth dilution method. Genomic characterization of MRSA ST398 strains was performed by WGS and their evolutionary relationships were assessed by phylogenetic analysis. RESULTS: Two LA-MRSA ST398 isolates were recovered from bulk tank milk samples in two geographically distant farms in China. Whole-genome analysis strongly suggested that the LA-MRSA ST398 strains were closely related to the highly virulent hospital-associated MRSA (HA-MRSA) ST398 strains in China. CONCLUSIONS: The presence of LA-MRSA ST398 in bulk tank milk might be a serious threat to public health, highlighting the need for active surveillance of LA-MRSA in healthy cattle in China.

An ShK-domain serine protease of Eriocheir sinensis regulates the PO activity to resist Spiroplasma eriocheiris infection.

A novel serine protease contains two ShK-domain was found from the Chinese mitten crab Eriocheir sinensis (EsShK-SP). The full-length EsShK-SP cDNA is 1927 bp and contains a 1260-bp open reading frame encoding a protein of 420 amino acids, including a signal peptide, two ShK domain, and Tryp-SPC domain. Quantitative real-time PCR showed that EsShK-SP was expressed mainly in the hemocytes, gills, intestine, and nerve, but weakly in heart, muscle, and hepatopancreas. After infected with Spiroplasma eriocheiris, the expression of EsShK-SP was significantly up-regulated from 1 d to 9 d. The Tryp-SPC domain was ligated with pGEX-4T-1 vector and prokaryotic expressed to obtain recombinant protein rSPC. When rSPC and S. eriocheiris stimulated the hemocytes of E. sinensis, the PO activity was significantly up-regulated. The subcellular localization revealed that recombinant EsShK-SP was mainly located in the cytoplasm of Drosophila S2 cells. Both absolute real-time PCR and confocal laser scanning microscope results showed that over-expression of EsShK-SP in S2 cells could decrease the copy number of S. eriocheiris. Meanwhile, the over-expression of EsShK-SP also increased the PO activity and cell viability of S2 cells. After EsShK-SP RNA interference using dsRNA, the expression levels of proPO and activity of PO decreased significantly from 48 h to 96 h. The knockdown of EsShK-SP by RNAi resulted in the copy number of S. eriocheiris in the EsShK-SP silenced group was significantly increased compared to the control groups during S. eriocheiris infection. Meanwhile, the survival rate of crabs decreased in the EsShK-SP-dsRNA group. The above results indicated that EsShK-SP plays an important immune role during E. sinensis against S. eriocheiris through regulation of the proPO system.

Achieving Stable Partial Nitritation in an Acidic Nitrifying Bioreactor.

Partial nitritation providing a suitable effluent for subsequent anammox is a critical step in a two-stage autotrophic nitrogen removal system. This study demonstrates an innovative approach for attaining partial nitritation in an acidic bioreactor operating at a slightly low pH (i.e., 5-6). This approach is based on our hypothesis in this study that acid-tolerant ammonia-oxidizing bacteria (AOB) can produce nitrite and protons to self-sustain free nitrous acid (FNA, NO2- + H+ ↔ HNO2) at a parts per million level, as an inhibitor of nitrite-oxidizing bacteria (NOB). With influent nitrogen of about 200 mg/L and operating conditions of high dissolved oxygen, long sludge retention time, and moderate temperature, a lab-scale acidic bioreactor with FNA up to 2 mg of HNO2-N/L successfully established stable nitrite accumulation in the effluent for 200 days, with an average ratio [NO2-/(NO2- + NO3-)] exceeding 95%. A 16S rRNA amplicon sequencing analysis showed that Nitrosospira was the dominant AOB in the biomass of the bioreactor, while Nitrosomonas and Nitrospira, two typical nitrifying genera in neutral wastewater treatment, both disappeared after the startup of partial nitritation. Kinetic characterization revealed that Nitrosospira had a substrate affinity of 11.4-16.5 mg of total ammonia (NH4+ + NH3)/L. It also revealed that less than 3.5 mg of HNO2-N/L FNA did not inhibit AOB activity significantly. Acidic operation is economically attractive because it can be achieved via acidophilic ammonia oxidation without adding chemical acid. However, hazardous gas, nitric oxide (NO), should be removed from gas produced by acidic nitrifying bioreactors.

Presence of NDM in non-E. coli Enterobacteriaceae in the poultry production environment.

OBJECTIVES: Characterization of non-Escherichia coli NDM-carrying Enterobacteriaceae in the poultry production environment. METHODS: A total of 36 NDM-positive Enterobacteriaceae (22 Klebsiella pneumoniae, 13 Enterobacter cloacae and 1 Salmonella enterica) were isolated from a chicken farm and WGS was conducted using Illumina Hiseq2500. The genomic characterization of the isolates acquired through WGS analysis included the genomic context-flanking blaNDM genes, MLST, the antibiotic resistance genes (ARGs) and replicon types of plasmids. WGS information for another 73 K. pneumoniae isolates from different sources was retrieved from GenBank and then combined with isolates in this study for comparative genomic and phylogenetic analysis. RESULTS: Three types of genetic environment carrying blaNDM were identified in 36 non-E. coli Enterobacteriaceae isolates. Sequence comparison analysis indicated these genetic environments were completely identical to our previous findings. WGS further revealed three major types of plasmids (IncFIB, IncX3 and IncFII) from these isolates and the phylogenetic analysis suggested several K. pneumoniae isolates with ST11, ST37 and ST147 from the commercial chicken farm that were closely related to isolates of human origin. CONCLUSIONS: The blaNDM-harbouring genetic contexts were identified not only in E. coli, but also in K. pneumoniae, E. cloacae and S. enterica, which may indicate that blaNDM has been widely disseminated to non-E. coli Enterobacteriaceae species in animal farms. The close relationship of K. pneumoniae isolates from different origins suggests they could serve as a key vehicle for the transfer of ARGs between humans and food animal production environments.

Mobile colistin resistance gene mcr-5 in porcine Aeromonas hydrophila.

Objectives: To characterize the mobile colistin resistance gene mcr-5 in Aeromonas hydrophila from backyard pigs in rural areas of China. Methods: Pig faecal samples from 194 households were directly tested for the presence of mcr-5 by PCR assay and the phenotypic antimicrobial susceptibility profiles of the mcr-5-positive isolates were determined using the broth dilution method. The genomic location and transferability of mcr-5 were analysed by S1-PFGE with Southern blotting and DNA hybridization, and natural transformation, respectively. One strain isolated from an mcr-5-positive sample was subjected to WGS and the stability of the mcr-5-harbouring plasmid over successive generations was examined by subculturing. Results: One mcr-5-positive A. hydrophila isolate showing resistance, with a colistin MIC of 4 mg/L, was isolated from a backyard pig faecal sample. mcr-5 was located on a 7915 bp plasmid designated pI064-2, which could naturally transform into a colistin-susceptible A. hydrophila strain of porcine origin and mediated colistin resistance in both the original isolate and its transformants. The plasmid backbone (3790 bp) of pI064-2 showed 81% nucleotide sequence identity to the corresponding region of the ColE2-type plasmid pAsa1 from Aeromonas salmonicida, while similar replication primases are widely distributed among aeromonads, Enterobacteriaceae and Pseudomonas species. Conclusions: To the best of our knowledge, this is the first identification of the novel colistin resistance gene mcr-5 in an A. hydrophila isolate from the faeces of a backyard pig. mcr-5 is expected to be able to disseminate among different bacterial species and genera.

A Real-Time Path Planning Algorithm for AUV in Unknown Underwater Environment Based on Combining PSO and Waypoint Guidance.

It is a challengeable task to plan multi-objective optimization paths for autonomous underwater vehicles (AUVs) in an unknown environments, which involves reducing travel time, shortening path length, keeping navigation safety, and smoothing trajectory. To address the above challenges, a real-time path planning approach combining particle swarm optimization and waypoint guidance is proposed for AUV in unknown oceanic environments in this paper. In this algorithm, a multi-beam forward looking sonar (FLS) is utilized to detect obstacles and the output data of FLS are used to produce those obstacles' outlines (polygons). Particle swarm optimization is used to search for appropriate temporary waypoints, in which the optimization parameters of path planning are taken into account. Subsequently, an optimal path is automatically generated under the guidance of the destination and these temporary waypoints. Finally, three algorithms, including artificial potential field and genic algorithm, are adopted in the simulation experiments. The simulation results show that the proposed algorithm can generate the optimal paths compared with the other two algorithms.

A Real-Time Reaction Obstacle Avoidance Algorithm for Autonomous Underwater Vehicles in Unknown Environments.

A novel real-time reaction obstacle avoidance algorithm (RRA) is proposed for autonomous underwater vehicles (AUVs) that must adapt to unknown complex terrains, based on forward looking sonar (FLS). To accomplish this algorithm, obstacle avoidance rules are planned, and the RRA processes are split into five steps Introduction only lists 4 so AUVs can rapidly respond to various environment obstacles. The largest polar angle algorithm (LPAA) is designed to change detected obstacle's irregular outline into a convex polygon, which simplifies the obstacle avoidance process. A solution is designed to solve the trapping problem existing in U-shape obstacle avoidance by an outline memory algorithm. Finally, simulations in three unknown obstacle scenes are carried out to demonstrate the performance of this algorithm, where the obtained obstacle avoidance trajectories are safety, smooth and near-optimal.

Plasmid-Mediated Novel blaNDM-17 Gene Encoding a Carbapenemase with Enhanced Activity in a Sequence Type 48 Escherichia coli Strain.

Carbapenem-resistant Enterobacteriaceae (CRE) have spread worldwide, leaving very few treatment options available. New Delhi metallo-beta-lactamase (NDM) is the main carbapenemase mediating CRE resistance and is of increasing concern. NDM-positive Enterobacteriaceae of human origin are frequently identified; however, the emergence of NDM, and particularly novel variants, in bacteria of food animal origin has never been reported. Here, we characterize a novel NDM variant (assigned NDM-17) identified in a ß-lactam-resistant sequence type 48 (ST48) Escherichia coli strain that was isolated from a chicken in China. Compared to NDM-1, NDM-17 had three amino acid substitutions (V88L, M154L, and E170K) that confer significantly enhanced carbapenemase activity. Compared to NDM-5, NDM-17 had only one amino acid substitution (E170K) and slightly increased isolate resistance to carbapenem, as indicated by increased MIC values. The gene encoding NDM-17 (blaNDM-17) was located on an IncX3 plasmid, which was readily transferrable to recipient E. coli strain J53 by conjugation, suggesting the possibility of the rapid dissemination of blaNDM-17 Enzyme kinetics showed that NDM-17 could hydrolyze all ß-lactams tested, except for aztreonam, and had a significantly higher affinity for all ß-lactams tested than did NDM-5. The emergence of this novel NDM variant could pose a threat to public health because of its transferability and enhanced carbapenemase activity.

Presence of VIM-Positive Pseudomonas Species in Chickens and Their Surrounding Environment.

Metallo-ß-lactamase gene blaVIM was identified on the chromosome of four Pseudomonas sp. isolates from a chicken farm, including one Pseudomonas aeruginosa isolate from a swallow (Yanornis martini), one Pseudomonas putida isolate from a fly, and two P. putida isolates from chickens. The four isolates shared two variants of blaVIM-carrying genomic contexts that resemble the corresponding regions of clinical metallo-ß-lactamase-producing Pseudomonas spp. Our study suggests that the surveillance of carbapenemase-producing bacteria in livestock and their surrounding environment is urgently needed.

Removal and recovery of N, P and K from urine via ammonia stripping and precipitations of struvite and struvite-K.

This study investigated the recovery of N, P and K from source-separated urine in laboratory-scale combined processes of air stripping and precipitation. Two operation scenarios were carried out to recover N/P (named partial scenario) and N/P/K (named complete scenario). Most of the nutrients were recovered (>70%) by optimising the operation parameters including the increase of air flow rate and more dosages of the sources of Mg and P. Absorbent rich in ammonium sulphate and solid precipitates including struvite, struvite-K, and struvite-Na was produced. The simultaneous recovery of P and K was the key process to determine the substance input. The ratio of substance input to nutrient recovered (P2O5 and K2O) was 4.14 in the partial scenario, whereas it increased to 10.61 in the complete scenario. The inevitable co-precipitation of struvite-Na mainly responded for the lower economic efficiency of the complete scenario.

Complete ammonia oxidation (comammox) at pH 3-4 supports stable production of ammonium nitrate from urine.

This study developed a new process that stably produced ammonium nitrate (NH4NO3), an important and commonly used fertilizer, from the source-separated urine by comammox Nitrospira. In the first stage, the complete conversion of ammonium to nitrate was achieved by comammox Nitrospira. In this scenario, the pH was maintained at 6 by adding external alkali, which also provided sufficient alkalinity for full nitrification. In the second stage, the NH4NO3 was produced directly by comammox Nitropsira by converting half of the ammonium in urine into nitrate. In this case, no alkali was added and pH automatically dropped and self-maintained at an extremely acidic level (pH 3-4). In both scenarios, negligible nitrite accumulation was observed, while the final product of the second stage contained ammonium and nitrate at the molar ratio of 1:1. The dominance of comammox Nitrospira over canonical ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) was systematically proved by the combination of 16S rRNA gene amplicon sequencing, quantitative polymerase chain reaction, and metagenomics. Notably, metagenomic sequencing suggested that the relative abundance of comammox Nitrospira was over 20 % under the acidic condition at pH 3-4, while canonical AOB and NOB were undetectable. Batch experiments showed that the optimal pH for the enriched comammox Nitrospira was ∼7, which could sustain their activity in a wider pH range from 4 to 8 surprisingly but lost activity at pH 3 and 9. The findings not only present an application potential of comammox Nitrospira in nitrogen recovery from urine wastewater but also report the survivability of comammox bacteria in acidic environments.

Characterization of Extended-Spectrum ß-Lactamase-Producing Escherichia coli in Animal Farms in Hunan Province, China.

Multi-drug resistance of bacteria producing extended-spectrum ß-lactamase (ESBL) is a public health challenge. Thus, this study aimed to investigate the antimicrobial susceptibility of ESBL-producing Escherichia coli (ESBL-EC) in Hunan Province, China. A total of 1366 fecal samples were collected from pig, chicken, and cattle farms over a six-year period, which were assessed using strain isolation, 16S rRNA identification, polymerase chain reaction, drug sensitivity testing, whole-genome sequencing, and bioinformatics analysis. The results showed an overall prevalence of 6.66% for ESBL-EC strains, with ESBL positivity extents for pigs, chickens, and cattle isolates at 6.77%, 6.54%, and 12.5%, respectively. Most ESBL-EC isolates were resistant to cefotaxime, tetracycline, and trimethoprim-sulfamethoxazole; however, all the isolates were susceptible to meropenem, with relatively low resistance to amikacin and tigecycline. Various multi-locus sequence types with different origins and similar affinities were identified, with ST155 (n = 16) being the most common subtype. Several types of resistance genes were identified among the 91 positive strains, with beta-lactamase blaCTX-M-55 being the most common ESBL genotype. IncFIB was the predominant plasmid type. Widespread use of antibiotics in animal farming may increase antibiotic resistance, posing a serious threat to the health of farmed animals and, thus, to human food security and health.

Significance of Siglec-15 expression in colorectal cancer: association with advanced disease stage and fewer tumor-infiltrating lymphocytes.

Siglec-15, a novel immune suppressor, is upregulated in many human cancers. The aim of this study was to explore the expression of Siglec-15 in colorectal cancer (CRC), and investigate whether Siglec-15 could be a potential target for cancer immunotherapy in patients with CRC. We performed immunohistochemical analyses of Siglec-15 on a cohort of 805 patients with CRC and made comparisons between clinicopathological characteristics, PD-L1 expression, CD3, CD8, CD45RO tumor-infiltrating lymphocytes (TILs), and prognosis. We found that Siglec-15 expression was commonly detected in tumor cells (48.3%) and tumor-associated stromal cells (33.4%), and was more frequently observed than PD-L1 expression in tumor cells. In contrast, Siglec-15 expression was weakly and scarcely found in normal mucosa (13%). Siglec-15 overexpression in tumor cells was associated with advanced TNM stage (p = 0.020). Co-expression of Siglec-15 and PD-L1 in tumor cells was found in 14.4% of patients, and Siglec-15 expression was detected in almost half of PD-L1 negative cases. Elevated Siglec-15 expression in tumor and stromal cells was associated with sparser CD45RO and CD8 TILs (p = 0.035 and p = 0.004, respectively). The expression of Siglec-15 did not have prognostic significance. In summary, compared to PD-L1, Siglec-15 protein expression is more prevalent in CRC and is associated with advanced disease stage and fewer TILs. These findings support Siglec-15 as a potential cancer immunotherapy target, in addition to PD-1/PD-L1 inhibitors, in patients with CRC.

Characteristics of tet(X4)-Producing Escherichia coli in Chicken and Pig Farms in Hunan Province, China.

BACKGROUND: The plasmid-mediated tigecycline resistance gene tet(X4) confers a high level of resistance to tigecycline. The experiment aims to investigate the prevalence and characterization of tet(X4) in Escherichia coli isolates from chicken and pig farms in Hunan province, China. METHODS: A total of six tet(X4) positive strains were identified in 257 E. coli derived from chicken samples in Xiangtan city (n = 2), pig samples in Xiangxiang city (n = 1), Chenzhou city (n = 2), and Zhuzhou city (n = 1). The presence of tet(X4) was directly detected by PCR assay, and then the broth dilution method determined the antimicrobial susceptibility profile of the tet(X4)-positive isolates. Genomic locations were identified by whole-genome sequencing (WGS) and bioinformatics. RESULTS: Almost all tet(X4)-positive strains showed high resistance to multidrug, including tigecycline. Resistome analysis revealed many antibiotic resistance genes, including those with resistance to tetracyclines, ß-lactams, phenicols, quinolones, lincosamides chloramphenicol, aminoglycosides and sulfamids. These tet(X4)-bearing strains exhibited six distract STs, such as ST10, 202, ST218, ST362, ST2077, ST7068. The plasmid replicon types carrying tet(X4) were the hybrid plasmid IncFIA(HI1)/IncHIA/IncHIB(R27) (5/6) and IncX1 (1/6). CONCLUSIONS: The presence of similar genetic environments in E. coli from different cities suggests there may be horizontal transmission pathways promoting the broad spread of drug-resistant genes in Hunan Province, putting great pressure on multidrug resistance monitoring.