Correlation between the development of phage resistance and the original antibiotic resistance of host bacteria under the co-exposure of antibiotic and bacteriophage.

Bacteriophages (phages) are viruses capable of regulating the proliferation of antibiotic resistant bacteria (ARB). However, phages that directly cause host lethality may quickly select for phage resistant bacteria, and the co-evolutionary trade-offs under varying environmental conditions, including the presence of antibiotics, remains unclear as to their impact on phage and antibiotic resistance. Here, we report the emergence of phage resistance in three distinct E. coli strains with varying resistance to ß-lactam antibiotics, treated with different ampicillin (AMP) concentrations. Hosts exhibiting stronger antibiotic resistance demonstrated a higher propensity to develop and maintain stable phage resistance. When exposed to polyvalent phage KNT-1, the growth of AMP-sensitive E. coli K12 was nearly suppressed within 18 h, while the exponential growth of AMP-resistant E. coli TEM and super-resistant E. coli NDM-1 was delayed by 12 h and 8 h, respectively. The mutation frequency and mutated colony count of E. coli NDM-1 were almost unaffected by co-existing AMP, whereas for E. coli TEM and K12, these metrics significantly decreased with increasing AMP concentration from 8 to 50 µg/mL, becoming unquantifiable at 100 µg/mL. Furthermore, the fitness costs of phage resistance mutation and its impact on initial antibiotic resistance in bacteria were further examined, through analyzing AMP susceptibility, biofilm formation and EPS secretion of the isolated phage resistant mutants. The results indicated that acquiring phage resistance could decrease antibiotic resistance, particularly for hosts lacking strong antibiotic resistance. The ability of mutants to form biofilm contributes to antibiotic resistance, but the correlation is not entirely positive, while the secretion of extracellular polymeric substance (EPS), especially the protein content, plays a crucial role in protecting the bacteria from both antibiotic and phage exposure. This study explores phage resistance development in hosts with different antibiotic resistance and helps to understand the limitations and possible solutions of phage-based technologies.

Physicochemical and Functional Similarity Assessment Between Proposed Bevacizumab Biosimilar BAT1706 and Reference Bevacizumab.

BACKGROUND: BAT1706 is a proposed biosimilar of bevacizumab, a vascular endothelial growth factor A (VEGF-A)-targeting biologic used to treat several different cancers, including metastatic colorectal cancer. A comprehensive physicochemical and functional similarity assessment is a key component of demonstrating biosimilarity between a reference biologic and a proposed biosimilar. Here we report the physicochemical and functional similarity of BAT1706 and reference bevacizumab sourced from both the United States (US-bevacizumab) and the European Union (EU-bevacizumab). METHOD: A large range of product attributes, including primary and higher order structure, post-translational modifications, purity, stability, and potency, were characterized for BAT1706 and EU/US-bevacizumab using sensitive state-of-the-art analytical techniques. Up to 18 lots of US- and 29 lots of EU-bevacizumab, and 10 unique drug substance lots of BAT1706, were assessed. RESULT: BAT1706 was shown to have an identical amino acid sequence and an indistinguishable higher-order structure compared with EU/US-bevacizumab. BAT1706 and EU/US-bevacizumab also exhibited similar post-translational modifications, glycan profiles, and charge variants. Potency, assessed using a wide range of bioassays, was also shown to be comparable between BAT1706 and EU/US-bevacizumab, with statistical equivalence demonstrated for VEGF-A binding and neutralizing activity. CONCLUSION: Overall, this extensive comparability exercise demonstrated BAT1706 to match EU/US-bevacizumab in terms of all physicochemical and functional attributes assessed.

Effects of abiotic factors on the stability and infectivity of polyvalent coliphage.

Bacteriophage has attracted growing interest as a promising therapeutic agent for pathogenic bacteria, especially for antibiotic-resistant bacteria. However, the various abiotic conditions could impact the stability of phages and further threat host-virus interactions. Here, we investigated the stability and lytic activity of virulent polyvalent coliphage (named PE1) by double-layer plaque assay. PE1 can efficiently infect both the drug-sensitive Escherichia coli K12 and multidrug-resistant E. coli NDM-1 even after prolonged storage at 4 °C for up to two months. Results showed that PE1 exhibits an outstanding stability to infect E. coli strains under a wide range of thermal (4 °C-60 °C) and pH (4-11) conditions, which covers the thermal and pH variations of most wastewater treatment plants. Moreover, PE1 exhibited high resistibility to heavy metals exposure including Cu2+, Cd2+, Co2+, and Cr3+ at the concentrations below 0.5 mM, and an excellent resistant ability to the variation of ionic strength, which still retained strong infectious ability even treated with saturated sodium chloride solution (350 g/L). This work shows that polyvalent phage PE1 has a strong adaptive capacity to various abiotic factors and should be a good candidate of being an antibacterial agent, especially for antibiotic-resistant bacteria control in sewage.

Rapid preparation of 1-vinylimidazole based non-affinity polymers for the highly-selective purification of antibodies from multiple biological sources.

There is an urgent need for developing advanced purification techniques with the merits of low cost and satisfactory capacity in order to meet the challenges in the current downstream purification of monoclonal antibodies (mAbs). Herein, a simple and inexpensive nitrogen heterocycle molecule, 1-vinylimidazole (VIM), was proposed as the capture ligand of antibodies for the first time. The corresponding VIM-based non-affinity polymeric material (polyVIM) was then fabricated via a one-step polymerization for use in the highly selective purification of antibodies. Compared to the previously reported materials, this novel material exhibited many advantages without clearly sacrificing selectivity, such as a simpler and faster fabrication (within 1.5 h), comparable or even higher binding capacity (saturated static adsorption capacity > 190 mg/g polymer, dynamic binding capacity about 31.62 mg/g polymer), lower non-specific protein adsorption, and much lower cost. Notably, the polyVIM can effectively purify the antibodies from multiple biological sources with high purity (95.4% for mAbs in the cell culture medium, 93.3% for hIgG in the human serum), with an acceptable recovery (91.6% for mAbs, 77.0% for hIgG), and good reusability (> 10 times). Moreover, the target ELISA binding assay and NFAT-luc reporter gene assay demonstrated that the enriched antibodies can well maintain their binding activity and bioactivity during the whole purification process. The excellent performance of the polyVIM material may be attributed to the high recognition ability of VIM for antibodies, as well as the biocompatible and antifouling properties of the porous polymer. This study provides a promising alternative material for the purification of mAbs in downstream processes and the enrichment of hIgG in human serum.

Novel Butane-Oxidizing Bacteria and Diversity of bmoX Genes in Puguang Gas Field.

To investigate the diversity of butane-oxidizing bacteria in soils contaminated by long-term light hydrocarbon microseepage and the influence of butane on the soil microbial community, a quantitative study and identification of butane-oxidizing bacteria (BOB) in soils at the Puguang gas field were performed by DNA-based stable isotope probing (DNA-SIP). For the first time, two phylotypes corresponding to the genera Giesbergeria and Ramlibacter were identified as being directly involved in butane oxidation, in addition to the well-known light hydrocarbon degrader Pseudomonas. Furthermore, bmoX genes were strongly labeled by 13C-butane, and their abundances in gas field soils increased by 43.14-, 17.39-, 21.74-, and 30.14-fold when incubated with butane for 6, 9, 12, and 14 days, respectively, indicating that these bmoX-harboring bacteria could use butane as the sole carbon and energy source and they play an important role in butane degradation. We also found that the addition of butane rapidly shaped the bacterial community and reduced the diversity of bmoX genes in the gas field soils. These findings improve our understanding of BOB in the gas field environment and reveal the potential for their applications in petroleum exploration and bioremediation.

Biodegradation of BTEX Aromatics by a Haloduric Microbial Consortium Enriched from a Sediment of Bohai Sea, China.

This study focused on a haloduric BTEX-degrading microbial consortium EC20 enriched from Bohai Sea sediment. EC20 degraded 87% of BTEX at 435 mg L-1 initial concentration (benzene, toluene, ethylbenzene, and xylenes in equal proportions) in the presence of 3.4% NaCl. 16S rRNA gene-based PCR-DGGE profiles revealed that the dominant bacteria in EC20 were Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes at the phylum level, and Pseudomonas, Mesorhizobium, Achromobacter, Stenotrophomonas, and Halomonas at the genus level. PCR detection of genes coding the key enzymes which participated in BTEX degradation pathways showed that the enriched consortium EC20 contained TOL pathway and TOD pathway to initiate biodegradation of BTEX.

[Dioxin Pollution and Occupational Inhalation Exposure of PCDD/Fs in Municipal Solid Waste Incinerator].

To assess the pollution levels, characteristics, and the pollution sources and occupational inhalation exposure of polychlorinated dibenzo-p-dioxins and dibenzofurans(PCDD/Fs)in the workshops,ambient air samples in different types of incinerators of two municipal solid waste incinerators(MSWI) were collected and analyzed. The results showed that ① The I-TEQ concentration ranged from 0.034-2.152 pg·m-3in the two waste incineration plants, and the most sites' I-TEQ exceeded the ambient air quality standard. Besides, the I-TEQ concentration behind the incineration plant was higher than others. ② The dioxins in incineration plant were dominated by OCDD and 1,2,3,4,6,7,8-HpCDD. For MSWI A, the flue gas and the fly ash had major effect on PCDD/Fs, while the dioxins pollution in MSWI B was only affected by the fly ash. ③ Occupational inhalation exposure of PCDD/Fs was 0.01-1.10 pg·(kg·d)-1 in incineration plant, some occupational inhalation exposure values exceeded the evaluation standard, and the areas behind the incinerators were evaluated to have a high exposure risk.

Syntrophic Interactions Within a Butane-Oxidizing Bacterial Consortium Isolated from Puguang Gas Field in China.

Butane oxidation by the hydrocarbon degradation bacteria has long been described, but little is known about the microbial interaction in this process. To investigate this interaction, the efficiency of butane oxidation was estimated in monocultures and co-cultures of six strains of butane-oxidizing bacteria (BOB) and a butanol-oxidizing strain. Results showed that the butane degradation velocity was at least 26 times higher in the co-culture of the seven strains (228.50 nmol h(-1)) than in the six individual monocultures (8.71 nmol h(-1)). Gas chromatographic analysis of metabolites in the cultures revealed the accumulation of butanol in the monocultures of BOB strains but not in the co-culture with the butanol-oxidizing strain. These results evidenced a novel syntrophic association between BOB and butanol-oxidizing bacteria in the butane oxidation. The BOB strains oxidized butane into butanol, but this activity was inhibited by the accumulated butanol in monocultures, whereas the removal of butanol by the butanol-oxidizing strain in co-culture could eliminate the suppression and improve the butane degradation efficiency. In the co-culture, both BOB and butanol-oxidizing bacteria could grow and the time needed for butane complete removal was shortened from more than 192 h to less than 4 h. The unsuppressed effect of the co-culture was also consistent with the results of reverse transcription quantitative real-time PCR (RT-qPCR) of bmoX gene because increased expression of this gene was detected during the syntrophic growth compared with that in monoculture, pointing to the upregulation of bmoX in the syntrophic interaction.

A proposal of "core enzyme" bioindicator in long-term Pb-Zn ore pollution areas based on topsoil property analysis.

To study the effects of long-term mining activities on the agricultural soil quality of Mengnuo town in Yunnan province, China, the heavy metal and soil enzyme activities of soil samples from 47 sites were examined. The results showed that long-term mining processes led to point source heavy metal pollution and Pb, Cd, Zn and As were the primary metal pollutants. Polyphenoloxidase was found the most sensitive soil enzyme activity and significantly correlated with almost all the metals (P < 0.05). Amylase (for C cycling), acid phosphatase (for P cycling) and catalase (for redox reaction) activities showed significantly positive correlations (P < 0.05) with Pb, Cd, Zn and As contents. The correlations between soil enzymes activities and Cd, Pb and Zn contents were verified in microcosm experiments, it was found that catalase activity had significant correlations (P < 0.05) with these three metals in short-term experiments using different soils under different conditions. Based on both field investigation and microcosm simulation analysis, oxidoreductases activities (rather than a specific enzyme activity) were suggested to be used as "core enzyme", which could simply and universally indicate the heavy metal pollution degrees of different environments. And hydrolases (for C, N, P and S recycling) could be used as a supplement to improve correlation accuracy for heavy metal indication in various polluted environments.

Universal Indicators for Oil and Gas Prospecting Based on Bacterial Communities Shaped by Light-Hydrocarbon Microseepage in China.

Light hydrocarbons accumulated in subsurface soil by long-term microseepage could favor the anomalous growth of indigenous hydrocarbon-oxidizing microorganisms, which could be crucial indicators of underlying petroleum reservoirs. Here, Illumina MiSeq sequencing of the 16S rRNA gene was conducted to determine the bacterial community structures in soil samples collected from three typical oil and gas fields at different locations in China. Incubation with n-butane at the laboratory scale was performed to confirm the presence of "universal microbes" in light-hydrocarbon microseepage ecosystems. The results indicated significantly higher bacterial diversity in next-to-well samples compared with background samples at two of the three sites, which were notably different to oil-contaminated environments. Variation partitioning analysis showed that the bacterial community structures above the oil and gas fields at the scale of the present study were shaped mainly by environmental parameters, and geographic location was able to explain only 7.05% of the variation independently. The linear discriminant analysis effect size method revealed that the oil and gas fields significantly favored the growth of Mycobacterium, Flavobacterium, and Pseudomonas, as well as other related bacteria. The relative abundance of Mycobacterium and Pseudomonas increased notably after n-butane cultivation, which highlighted their potential as biomarkers of underlying oil deposits. This work contributes to a broader perspective on the bacterial community structures shaped by long-term light-hydrocarbon microseepage and proposes relatively universal indicators, providing an additional resource for the improvement of microbial prospecting of oil and gas.

Cloacibacterium rupense sp. nov., isolated from freshwater lake sediment.

A Gram-negative, yellow-pigmented bacterium, designated strain R2A-16(T), was isolated from sediment of Rupa Lake in Nepal and analysed using a polyphasic taxonomic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain R2A-16(T) is affiliated to the genus Cloacibacterium of the family Flavobacteriaceae; 16S rRNA gene sequence similarity between strain R2A-16(T) and Cloacibacterium normanense CCUG 46293(T) was 98.07 %. The isolate contained iso-C(15 : 0) (35.6 %) as the major fatty acid and menaquinone MK-6 as the predominant respiratory quinone. The G+C content of the genomic DNA was 33.3 mol%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain R2A-16(T) represents a novel species of the genus Cloacibacterium, for which the name Cloacibacterium rupense sp. nov. is proposed; the type strain is R2A-16(T) (=CGMCC 1.7656(T) =NBRC 104931(T)).

Flavobacterium cauense sp. nov., isolated from sediment of a eutrophic lake.

A Gram-negative, rod-shaped, yellow-pigmented bacterium, designated strain R2A-7(T), was isolated from sediment of the eutrophic Taihu Lake in Jiangsu Province, China. The isolate was subjected to a taxonomic study using a polyphasic approach. Phylogenetic analysis based on the 16S rRNA gene sequences placed strain R2A-7(T) within the genus Flavobacterium in the family Flavobacteriaceae. The highest sequence similarity was found with Flavobacterium saliperosum (98.3 %), followed by other Flavobacterium species with similarities <96.0 %. The major fatty acids (>5 %) were 15 : 0 iso, 17 : 1 iso omega9c, 17 : 0 iso 3-OH, 15 : 1 iso G and 15 : 0 iso 3-OH. The G+C content of the genomic DNA of strain R2A-7(T) was 37.7 mol%. The DNA-DNA relatedness value with F. saliperosum CGMCC 1.3801(T) was 40.6 %. Molecular and phenotypic data suggest that strain R2A-7(T) represents a novel species within the genus Flavobacterium, for which the name Flavobacterium cauense is proposed. The type strain is R2A-7(T) (=CGMCC 1.7270(T)=NBRC 104929(T)).