Advanced strategies for combating bacterial biofilms

Biofilms are communities of microorganisms that are formed on and attached to living or nonliving surfaces and are surrounded by an extracellular polymeric material. Biofilm formation enjoys several advantages over the pathogens in the colonization process of medical devices and patients' organs. Unlike planktonic cells, biofilms have high intrinsic resistance to antibiotics and sanitizers, and overcoming them is a significant problematic challenge in the medical and food industries. There are no approved treatments to specifically target biofilms. Thus, it is required to study and present innovative and effective methods to combat a bacterial biofilm. In this review, several strategies have been discussed for combating bacterial biofilms to improve healthcare, food safety, and industrial process.     

New insights into nanomaterials combating bacteria: ROS and beyond

<正>Nanozymes are nanomaterials-based enzyme mimics which have received increasing research interests owing to their distinctive merits over natural enzymes, such as low costs,facile preparation, and high stability (Wei and Wang, 2013;Gao and Yan, 2016). By now, a plenty of nanomaterials, like metal oxide nanoparticles, noble metal nanoparticles, nanocarbon materials and transition metal dichalcogenides have been reported to exhibit several enzyme-like catalytic activities (Sun et al., 2018)(Figure 1). Over the past decade,     

Enrichment strategies for nitrile-hydrolysing bacteria

A series of enrichments with different nitriles as sole source of nitrogen was performed in order to obtain a relationship between the selective nitrogen source and (i) the enzyme systems that are synthesized by the isolates and (ii) the enzyme specificities for the utilization of the nitriles. Bacteria were enriched with 2-phenylpropionitrile, 2-(2-methoxyphenyl)propionitrile, 2-phenylbutyronitrile, ibuprofen nitrile, naproxen nitrile, ketoprofen nitrile, ketoprofen amide, benzonitrile, or naphthalenecarbonitrile as sole nitrogen source and succinate as sole source of carbon and energy. 2-Phenylpropionitrile as nitrogen source resulted predominantly in the enrichment of gram-negative bacteria, which harboured nitrilase and in some cases also amidase activity. In contrast, with the other nitriles used, a substantial majority of gram-positive strains, mainly of the genus Rhodococcus, were isolated. These strains contained predominantly a nitrile hydratase/amidase system. The nitrilases and nitrile hydratases showed R or S selectivity with generally poor optical yields. In contrast, the amidases were almost exclusively S-selective, often forming the optically pure acids with an enantiomeric excess above 99%. The conversion of different nitriles by the isolates was compared. The nitrile-hydrolysing systems of the new isolates usually showed high activity against those nitriles that were used for the enrichment of the bacteria. Received: 13 November 1996 / Received revision: 4 February 1997 / Accepted: 10 February 1997     

New approaches to combating antimicrobial drug resistance

Recent work shows that the inhibition of the SOS stress response in Escherichia coli reduces the development of resistance to the antibiotics ciprofloxacin and rifampicin. This finding may help in the battle against the rise of resistance to antimicrobial drugs.     

Bactericidal effect of silver nanoparticles against multidrug-resistant bacteria

Infections caused by drug-resistant microorganisms result in significant increases in mortality, morbidity, and cost related to prolonged treatments. The antibacterial activity of silver nanoparticles against some drug-resistant bacteria has been established, but further investigation is needed to determine whether these particles could be an option for the treatment and prevention of drug-resistant microbial infections. Hence, we challenged different drug-resistant pathogens of clinical importance (multidrug-resistant Pseudomonas aeruginosa, ampicillin-resistant Escherichia coli O157:H7 and erythromycin-resistant Streptococcus pyogenes) with a suspension of silver nanoparticles. By means of a luciferase-based assay, it was determined that silver nanoparticles (1) inactivate a panel of drug-resistant and drug-susceptible bacteria (Gram positive and Gram negative), (2) exert their antibacterial activity through a bactericidal rather than bacteriostatic mechanism, and (3) inhibit the bacterial growth rate from the time of first contact between the bacteria and the nanoparticles. Additionally, strains with a resistant phenotype to silver nanoparticle were developed and used to explore the bactericidal mode of action of silver nanoparticles. Through a Kirby–Bauer test, it was shown that silver nanoparticles’ general mechanism of bactericidal action is based on inhibition of cell wall synthesis, protein synthesis mediated by the 30s ribosomal subunit, and nucleic acid synthesis. Our data suggest that silver nanoparticles are effective broad-spectrum biocides against a variety of drug-resistant bacteria, which makes them a potential candidate for use in pharmaceutical products and medical devices that may help to prevent the transmission of drug-resistant pathogens in different clinical environments.     

Systematic research to overcome newly emerged multidrug-resistant bacteria

In the 1980s, I found that the chromosomal β-lactamase of Klebsiella pneumoniae LEN-1 showed a very high similarity to the R-plasmid-mediated penicillinase TEM-1 on the amino acid sequence level, and this strongly suggested the origination of TEM-1 from the chromosomal penicillinases of K. pneumoniae or related bacteria. Moreover, the chromosomal K1 β-lactamase (KOXY) of Klebsiella oxytoca was found to belong to the class A β-lactamases that include LEN-1 and TEM-1, although KOXY can hydrolyze cefoperazone (CPZ) like the chromosomal AmpC-type cephalosporinases of various Enterobacteriaceae that can hydrolyze several cephalosporins including CPZ. Furthermore, my collaborators and I found plural novel serine-type β-lactamases, such as MOX-1, SHV-24, TEM-91, CTX-M-64, CMY-9, CMY-19, GES-3, GES-4, and TLA-3, mediated by plasmids. Besides these serine-type β-lactamases, we also first identified exogenously acquired metallo-β-lactamases (MBLs), IMP-1 and SMB-1, in imipenem-resistant Serratia marcescens, and the IMP-1-producing S. marcescens TN9106 became the index case for carbapenemase-producing Enterobacteriaceae. I developed the sodium mercaptoacetic acid (SMA)-disk test for the simple identification of MBL-producing bacteria. We were also the first to identify a variety of plasmid-mediated 16S ribosomal RNA methyltransferases, RmtA, RmtB, RmtC, and NpmA, from various Gram-negative bacteria that showed very high levels of resistance to a wide range of aminoglycosides. Furthermore, we first found plasmid-mediated quinolone efflux pump (QepA) and fosfomycin-inactivating enzymes (FosA3 and FosK). We also first characterized penicillin reduced susceptible Streptococcus agalactiae, macrolide-resistant Mycoplasma pneumoniae, as well as Campylobacter jejuni, and Helicobacter pylori, together with carbapenem-resistant Haemophilus influenzae. We constructed a PCR-based open reading frame typing method for rapid identification of Acinetobacter baumannii international clones.     

Bacteriophage isolated from non-target bacteria demonstrates broad host range infectivity against multidrug-resistant bacteria

Antibiotic resistance represents a global health challenge. The emergence of multidrug-resistant (MDR) bacteria such as uropathogenic Escherichia coli (UPEC) has attracted significant attention due to increased MDR properties, even against the last line of antibiotics. Bacteriophage, or simply phage, represents an alternative treatment to antibiotics. However, phage applications still face some challenges, such as host range specificity and development of phage resistant mutants. In this study, using both UPEC and non-UPEC hosts, five different phages were isolated from wastewater. We found that the inclusion of commensal Escherichia coli as target hosts during screening improved the capacity to select phage with desirable characteristics for phage therapy. Whole-genome sequencing revealed that four out of five phages adopt strictly lytic lifestyles and are taxonomically related to different phage families belonging to the Myoviridae and Podoviridae. In comparison to single phage treatment, the application of phage cocktails targeting different cell surface receptors significantly enhanced the suppression of UPEC hosts. The emergence of phage-resistant mutants after single phage treatment was attributed to mutational changes in outer membrane protein components, suggesting the potential receptors recognized by these phages. The findings highlight the use of commensal E. coli as target hosts to isolate broad host range phage with infectivity against MDR bacteria.     

The role of vaccines in combating antimicrobial resistance (AMR) bacteria

Most pathogens have developed an intrinsic capacity to thrive by developing resistance to antimicrobial compounds utilized in treatment. Antimicrobial resistance arises when microbial agents such as bacteria, viruses, fungi, and parasites alter their behaviour to make current conventional medicines inefficient. Vaccination is one of the most effective strategies to fight antimicrobial resistance. Vaccines, unlike drugs, are less likely to produce resistance since they are precise to their target illnesses. Vaccines against infectious agents such as Streptococcus pneumoniae and Haemophilus influenzae have already been shown to reduce tolerance to antimicrobial medications; however, vaccines against some antimicrobial-resistant pathogens such as Vibrio cholerae, Salmonella typhi, Escherichia coli, nosocomial infections, and pulmonary and diarrheal disease viruses require more research and development. This paper describes vaccine roles in combatting antimicrobial resistance, quantifies the overall advantages of vaccination as an anti-antimicrobial resistance approach, analyzes existing antimicrobial vaccines and those currently under development, and emphasizes some of the obstacles and prospects of vaccine research and development.     

Silver nanoparticles: the powerful nanoweapon against multidrug-resistant bacteria

In the present scenario, pharmaceutical and biomedical sectors are facing the challenges of continuous increase in the multidrug-resistant (MDR) human pathogenic microbes. Re-emergence of MDR microbes is facilitated by drug and/or antibiotic resistance, which is acquired way of microbes for their survival and multiplication in uncomfortable environments. MDR bacterial infections lead to significant increase in mortality, morbidity and cost of prolonged treatments. Therefore, development, modification or searching the antimicrobial compounds having bactericidal potential against MDR bacteria is a priority area of research. Silver in the form of various compounds and bhasmas have been used in Ayurveda to treat several bacterial infections since time immemorial. As several pathogenic bacteria are developing antibiotic resistance, silver nanoparticles are the new hope to treat them. This review discusses the bactericidal potential of silver nanoparticles against the MDR bacteria. This multiactional nanoweapon can be used for the treatment and prevention of drug-resistant microbes.     

Molecular strategies for overcoming antibiotic resistance in bacteria

Overuse of antibiotics in humans and livestock has led to the rapid evolution of bacteria that are resistant to multiple drugs such that even vancomycin, the drug of last resort, is no longer effective against some strains. Apart from the discovery and exploitation of the natural peptide antimicrobial agents that form part of the innate immune systems of plants and animals, there have been few new antibiotics developed in recent years. Here we review strategies designed to exploit recent advances in molecular biology, including recombinant DNA technology, molecular modelling and genomics to develop new antibacterial agents that overcome antibiotic resistance.     

New agents for Gram-positive bacteria

Infections caused by multiple-resistant Gram-positive organisms continue to occur at an alarming rate worldwide. Two new and unique antimicrobial agents targeted specifically against such organisms, quinupristin/dalfopristin and linezolid, have been approved for use in the USA in the past year and will play an important role in the treatment of life-threatening infections. In addition, several new fluoroquinolones have been approved recently or will be available in the near future to aid in the treatment of infections caused by resistant strains of Streptococcus pneumoniae.     

Biocontainment strategies for live lactic acid bacteria vaccine vectors

Stability is an important issue when engineering bacteria for use as live vaccine vectors. For the majority of live bacterial vaccines, the antigen-encoding gene is either plasmid located or integrated into the chromosome. Regardless, several safety concerns can be raised for both instances. One concern when using plasmid-encoded antigens is the transfer of antibiotic resistance markers. Alternatively, for chromosomal integrated antigens however, the concern focuses on the spread and possible release of genetically-modified microorganisms (GMM) into the environment, which is problematic. Their recombinant nature calls for a proper bio-containment strategy to be implemented or in place before any realistic attempt at releasing a live bacterial vaccine. No examples of human bacterial vaccines causing problems among animals have been found in the literature but the possibility exists and has to be both tested and evaluated before release of a live bacterial vaccine. The ideal GMM for use in humans should therefore contain the minimal amount of foreign DNA and must not include an antibiotic resistance marker. Furthermore, the possibilities of transgene horizontal transfer must be minimized, and GMM lethality for biocontainment should be achieved in an unconfined environment.     

小分子功能化的金纳米粒子对革兰氏阴性多药耐药细菌的抗菌特性

【目的】研究大肠杆菌tRNA合成底物类似物4,6-二氨基-2-巯基嘧啶功能化的金纳米粒子(gold nanoparticles,AuNPs)对革兰氏阴性多药耐药细菌的抗菌特性。【方法】以4,6-二氨基-2-巯基嘧啶为表面配体合成AuNPs,采用肉汤稀释法测定其对4种临床分离的革兰氏阴性多药耐药细菌的最低抑菌浓度(MIC)。通过不同浓度AuNPs处理后经平板计数绘制不同菌株的时间-杀菌动力学曲线。以铜绿假单胞菌为代表菌株,采用激光共聚焦显微镜、透射电子显微镜和凝胶电泳分析AuNPs对细菌细胞组分的损伤。通过亚致死浓度反复诱导评估细菌对AuNPs的耐药性演化。并以MTT实验初步评估了AuNPs对哺乳动物细胞的生物相容性。【结果】4,6-二氨基-2-巯基嘧啶介导的AuNPs平均粒径为6.8nm,zeta电位为+38.4mV。该AuNPs对4种临床分离的革兰氏阴性多药耐药细菌均表现出时间和浓度依赖的抗菌活性,MIC值介于4–8μg/mL之间。抗菌机制研究显示AuNPs主要通过诱导细菌细胞膜损伤和DNA断裂导致细菌死亡。耐药性演化评估发现细菌在为期30d的反复诱导下也基本不会对该AuNPs产生耐...     

Combination therapies for combating antimicrobial resistance

New drug development strategies are needed to combat antimicrobial resistance. The object of this perspective is to highlight one such strategy: treating infections with sets of drugs rather than individual drugs. We will highlight three categories of combination therapy: those that inhibit targets in different pathways; those that inhibit distinct nodes in the same pathway; and those that inhibit the very same target in different ways. We will then consider examples of naturally occurring combination therapies produced by micro-organisms, and conclude by discussing key opportunities and challenges for making more widespread use of drug combinations.     

Biofortification for combating 'hidden hunger' for iron

Micronutrient deficiencies are responsible for so-called 'hidden undernutrition'. In particular, iron (Fe) deficiency adversely affects growth, immune function and can cause anaemia. However, supplementation of iron can exacerbate infectious diseases and current policies of iron therapy carefully evaluate the risks and benefits of these interventions. Here we review the approaches of biofortification of valuable crops for reducing 'hidden undernutrition' of iron in the light of the latest nutritional and medical advances. The increase of iron and prebiotics in edible parts of plants is expected to improve health, whereas the reduction of phytic acid concentration, in crops valuable for human diet, might be less beneficial for the developed countries, or for the developing countries exposed to endemic infections.