Intrinsic Antibiotic Resistance Research Articles

Prevalence and Therapies of Antibiotic-Resistance in Staphylococcus aureus.

Infectious diseases are the second most important cause of human death worldwide; Staphylococcus aureus (S. aureus) is a very common human pathogenic microorganism that can trigger a variety of infectious diseases, such as skin and soft tissue infections, endocarditis, osteomyelitis, bacteremia, and lethal pneumonia. Moreover, according to the sensitivity to antibiotic drugs, S. aureus can be divided into methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA). In recent decades, due to the evolution of bacteria and the abuse of antibiotics, the drug resistance of S. aureus has gradually increased, the infection rate of MRSA has increased worldwide, and the clinical anti-infective treatment for MRSA has become more difficult. Accumulating evidence has demonstrated that the resistance mechanisms of S. aureus are very complex, especially for MRSA, which is resistant to many kinds of antibiotics. Therefore, understanding the drug resistance of MRSA in a timely manner and elucidating its drug resistance mechanism at the molecular level are of great significance for the treatment of S. aureus infection. A large number of researchers believe that analyzing the molecular characteristics of S. aureus can help provide a basis for designing effective prevention and treatment measures against hospital infections caused by S. aureus and further monitor the evolution of S. aureus. This paper reviews the research status of MSSA and MRSA, the detailed mechanisms of the intrinsic antibiotic resistance and the acquired antibiotic resistance, the advanced research on anti-MRSA antibiotics and novel therapeutic strategies for MRSA treatment.

A Glycosylated Cationic Block Poly(β-peptide) Reverses Intrinsic Antibiotic Resistance in All ESKAPE Gram-Negative Bacteria.

Carbapenem-resistant Gram-negative bacteria (GNB) are heading the list of pathogens for which antibiotics are the most critically needed. Many antibiotics are either unable to penetrate the outer-membrane or are excluded by efflux mechanisms. Here, we report a cationic block β-peptide (PAS8-b-PDM12) that reverses intrinsic antibiotic resistance in GNB by two distinct mechanisms of action. PAS8-b-PDM12 does not only compromise the integrity of the bacterial outer-membrane, it also deactivates efflux pump systems by dissipating the transmembrane electrochemical potential. As a result, PAS8-b-PDM12 sensitizes carbapenem- and colistin-resistant GNB to multiple antibiotics in vitro and in vivo. The β-peptide allows the perfect alternation of cationic versus hydrophobic side chains, representing a significant improvement over previous antimicrobial α-peptides sensitizing agents. Together, our results indicate that it is technically possible for a single adjuvant to reverse innate antibiotic resistance in all pathogenic GNB of the ESKAPE group, including those resistant to last resort antibiotics.

A Glycosylated Cationic Block Poly(β‐peptide) Reverses Intrinsic Antibiotic Resistance in All ESKAPE Gram‐Negative Bacteria

AbstractCarbapenem‐resistant Gram‐negative bacteria (GNB) are heading the list of pathogens for which antibiotics are the most critically needed. Many antibiotics are either unable to penetrate the outer‐membrane or are excluded by efflux mechanisms. Here, we report a cationic block β‐peptide (PAS8‐b‐PDM12) that reverses intrinsic antibiotic resistance in GNB by two distinct mechanisms of action. PAS8‐b‐PDM12 does not only compromise the integrity of the bacterial outer‐membrane, it also deactivates efflux pump systems by dissipating the transmembrane electrochemical potential. As a result, PAS8‐b‐PDM12 sensitizes carbapenem‐ and colistin‐resistant GNB to multiple antibiotics in vitro and in vivo. The β‐peptide allows the perfect alternation of cationic versus hydrophobic side chains, representing a significant improvement over previous antimicrobial α‐peptides sensitizing agents. Together, our results indicate that it is technically possible for a single adjuvant to reverse innate antibiotic resistance in all pathogenic GNB of the ESKAPE group, including those resistant to last resort antibiotics.

Novel Bacteriophages Capable of Disrupting Biofilms From Clinical Strains of Aeromonas hydrophila.

The increase in global warming has favored growth of a range of opportunistic environmental bacteria and allowed some of these to become more pathogenic to humans. Aeromonas hydrophila is one such organism. Surviving in moist conditions in temperate climates, these bacteria have been associated with a range of diseases in humans, and in systemic infections can cause mortality in up to 46% of cases. Their capacity to form biofilms, carry antibiotic resistance mechanisms, and survive disinfection, has meant that they are not easily treated with traditional methods. Bacteriophage offer a possible alternative approach for controlling their growth. This study is the first to report the isolation and characterization of bacteriophages lytic against clinical strains of A. hydrophila which carry intrinsic antibiotic resistance genes. Functionally, these novel bacteriophages were shown to be capable of disrupting biofilms caused by clinical isolates of A. hydrophila. The potential exists for these to be tested in clinical and environmental settings.

Phylogenetic analysis reveals an ancient gene duplication as the origin of the MdtABC efflux pump.

The efflux pumps from the Resistance-Nodulation-Division family, RND, are main contributors to intrinsic antibiotic resistance in Gram-negative bacteria. Among this family, the MdtABC pump is unusual by having two inner membrane components. The two components, MdtB and MdtC are homologs, therefore it is evident that the two components arose by gene duplication. In this paper, we describe the results obtained from a phylogenetic analysis of the MdtBC pumps in the context of other RNDs. We show that the individual inner membrane components (MdtB and MdtC) are conserved throughout the Proteobacterial species and that their existence is a result of a single gene duplication. We argue that this gene duplication was an ancient event which occurred before the split of Proteobacteria into Alpha-, Beta- and Gamma- classes. Moreover, we find that the MdtABC pumps and the MexMN pump from Pseudomonas aeruginosa share a close common ancestor, suggesting the MexMN pump arose by another gene duplication event of the original Mdt ancestor. Taken together, these results shed light on the evolution of the RND efflux pumps and demonstrate the ancient origin of the Mdt pumps and suggest that the core bacterial efflux pump repertoires have been generally stable throughout the course of evolution.

A potential antifungal and growth-promoting bacterium Bacillus sp. KTMA4 from tomato rhizosphere

Plant growth-promoting rhizobacteria are indigenous beneficial bacteria that will enhance plant growth as well as suppress phytopathogens. In the present study, the isolate KTMA4 showed the highest inhibition against major phytopathogens of tomato; Fusarium oxysporum (66%) and Alternaria solani (54%) after seven days of incubation. Analysis of the 16S rRNA gene sequence revealed that the isolate KTMA4 is Bacillus cereus (MG547975). The isolate produced in vitro plants growth-promoting factors such as Indole-3-acetic acid, ammonia, catalase, siderophore and 1-aminocyclopropane-1-carboxylate deaminase and it has nitrogen fixation ability. The bacterial strain has also produced lytic enzymes such as amylase, cellulase, xylanase, lipase, and protease. Moreover, the bacterium Bacillus cereus KTMA4 effectively produced biofilm, biosurfactants and salt-tolerant (5% NaCl). The bacterium exhibited intrinsic antibiotic resistance. The in vivo studies using tomato plants grown from seeds treated with the bacterial strain KTMA4 demonstrated an enhancement in seed germination percentage (86.66 ± 2.88) and vigour index (637.5 ± 21.65) over the uninoculated control (germination percentage- 28.33 ± 2.88 and vigour index- 42.5 ± 4.33). 60 days of greenhouse study revealed that the bacterial isolate enhanced the plant growth significantly (P ≤ 0.05) compared to the uninoculated control and the treated plants. Therefore the study suggests that the newly isolated rhizosphere bacterial strain can be used as a potential biocontrol agent against a multitude of fungal pathogens as well as a biofertilizer inoculant for tomato cultivation.

Cryo-electron Microscopy Structure of the Acinetobacter baumannii 70S Ribosome and Implications for New Antibiotic Development.

Antimicrobial resistance is a major health threat as it limits treatment options for infection. At the forefront of this serious issue is Acinetobacter baumannii, a Gram-negative opportunistic pathogen that exhibits the remarkable ability to resist antibiotics through multiple mechanisms. As bacterial ribosomes represent a target for multiple distinct classes of existing antimicrobial agents, we here use single-particle cryo-electron microscopy (cryo-EM) to elucidate five different structural states of the A. baumannii ribosome, including the 70S, 50S, and 30S forms. We also determined interparticle motions of the 70S ribosome in different tRNA bound states using three-dimensional (3D) variability analysis. Together, our structural data further our understanding of the ribosome from A. baumannii and other Gram-negative pathogens and will enable structure-based drug discovery to combat antibiotic-resistant bacterial infections.IMPORTANCEAcinetobacter baumannii is a severe nosocomial threat largely due to its intrinsic antibiotic resistance and remarkable ability to acquire new resistance determinants. The bacterial ribosome serves as a major target for modern antibiotics and the design of new therapeutics. Here, we present cryo-EM structures of the A. baumannii 70S ribosome, revealing several unique species-specific structural features that may facilitate future drug development to combat this recalcitrant bacterial pathogen.

Toward a structome of Acinetobacter baumannii drug targets.

Acinetobacter baumannii is well known for causing hospital-associated infections due in part to its intrinsic antibiotic resistance as well as its ability to remain viable on surfaces and resist cleaning agents. In a previous publication, A. baumannii strain AB5075 was studied by transposon mutagenesis and 438 essential gene candidates for growth on rich-medium were identified. The Seattle Structural Genomics Center for Infectious Disease entered 342 of these candidate essential genes into our pipeline for structure determination, in which 306 were successfully cloned into expression vectors, 192 were detectably expressed, 165 screened as soluble, 121 were purified, 52 crystalized, 30 provided diffraction data, and 29 structures were deposited in the Protein Data Bank. Here, we report these structures, compare them with human orthologs where applicable, and discuss their potential as drug targets for antibiotic development against A. baumannii.

Selected autochthonous rhizobia, applied in combination with AM fungi, improve seed quality of common bean cultivated in reduced fertilization condition

Phaseolus vulgaris cv. Billò is an important crop, widely cultivated in Western Piedmont (Italy). It can form symbioses both with arbuscular mycorrhizal fungi (AMF) and bacteria belonging to the Rhizobium genus: AMF can improve plant nutrition and fruit/seed quality, whereas R. leguminosarum is a Gram-negative bacterium able to fix atmospheric nitrogen. In this study, 15 Rhizobium strains, isolated either from root nodules or soil, were characterized by molecular and biochemical point of view (P-solubilization, siderophore and IAA production, intrinsic antibiotic resistance) and one of the selected strains was used for field trial. The aims were: i) to select highly efficient symbiotic bacteria adapted to local conditions and ii) to assess bean seed yield and quality, in response to AMF/rhizobia inoculation at reduced fertilization level. At harvest, yield (pod and seed number and weight) and seed quality (starch, protein, fiber and mineral concentrations), root nodulation and mycorrhizal colonization were measured. Inoculation did not influence yield and fiber concentration. On the contrary, the total seed protein amount was higher in the seeds of rhizobium-inoculated plants in combination or without AMF than in uninoculated plants. The starch content in seeds was increased by rhizobia inoculation. Mg, K and Zn seed accumulation was positively affected by AMF, while the Mn concentration was higher in the presence of rhizobia. In conclusion, plant inoculation strongly affected seed characteristics, improving bean quality also in conditions of reduced fertilization.

Clonal Diversity, Biofilm Formation, and Antimicrobial Resistance among Stenotrophomonas maltophilia Strains from Cystic Fibrosis and Non-Cystic Fibrosis Patients.

The intrinsic antibiotic resistance of Stenotrophomonas maltophilia, along with its ability to form biofilm both on abiotic surfaces and host tissues, dramatically affects the efficacy of the antibiotic therapy. In this work, 85 S. maltophilia strains isolated in several hospital of central Italy and from several clinical settings were evaluated for their genetic relatedness (by pulsed-field gel electrophoresis, PFGE), biofilm formation (by microtiter plate assay), and planktonic antibiotic resistance (by Kirby–Bauer disk diffusion technique). The S. maltophilia population showed a high genetic heterogeneity: 64 different PFGE types were identified, equally distributed in cystic fibrosis (CF) and non-CF strains, and some consisted of multiple strains. Most of the strains (88.2%) were able to form biofilm, although non-CF strains were significantly more efficient than CF strains. CF strains produced lower biofilm amounts than non-CF strains, both those from respiratory tracts and blood. Non-CF PFGE types 3 and 27 consisted of strong-producers only. Cotrimoxazole and levofloxacin were the most effective antibiotics, being active respectively against 81.2% and 72.9% of strains. CF strains were significantly more resistant to piperacillin/tazobactam compared to non-CF strains (90% versus 53.3%), regardless of sample type. Among respiratory strains, cotrimoxazole was more active against non-CF than CF strains (susceptibility rates: 86.7% versus 75%). The multidrug resistant phenotype was significantly more prevalent in CF than non-CF strains (90% versus 66.7%). Overall, the multidrug-resistance level was negatively associated with efficiency in biofilm formation. Our results showed, for the first time, that in S. maltophilia both classical planktonic drug resistance and the ability of biofilm formation might favor its dissemination in the hospital setting. Biofilm formation might in fact act as a survival mechanism for susceptible bacteria, suggesting that clinical isolates should be routinely assayed for biofilm formation in diagnostic laboratories.

Isolation, Screening and Evaluation of Multifunctional Strains of High Efficient Phosphate Solubilizing Microbes from Rhizosphere Soil

Phosphorus is one of the essential macronutrients which supports plant growth and is obtained from the soil. Phosphate solubilizing micro-organisms helps in solubilizing the insoluble form of phosphates into soluble forms. The present study is aimed to isolate phosphate solubilizing microorganisms from rhizosphere of four plants. P-solubilizing organisms were screened using the Pikovskaya's medium with Tricalcium phosphate as the sole source of phosphate. P-solubilization activity was measured in NBRIP medium and two fungal and bacterial strains obtained from the Colocasia esculenta were selected for their maximum solubilization indices. The selected strains were subjected for morphological, biochemical and genotypic identification and the strains were identified to be Pestalotiopsis microspora, Aquabacterium commune, Bacillus spp., and Aspergillus spp. All the identified strains were subjected to screening against various environmental stresses such as salinity, pH and temperatures. The strains were also tested for intrinsic antibiotic resistance. Finally, the strains were tested for the pesticide tolerance activity against 0.25% Cypermethrin. From these tests, it was observed that the strains were tolerant to salt concentration upto 4%, to temperature upto 37ᵒC. and to pH of 7.8. The strains were tested for the indole acetic acid production activity and were found to be positive for the test. The strains were found to resistant against all the antibiotics except Gentamicin and Co-Trimoxazole. It was also found that the strains were tolerant towards the 0.25% Cypermethrin upto 1g/L. The identified strains were further analyzed by the molecular studies and constructed phylogenetic tree analysis.

Clinical Outcomes of Clofazimine Use for Rapidly Growing Mycobacterial Infections

Abstract Background Rapidly growing mycobacteria (RGM) have high rates of intrinsic antibiotic resistance and require prolonged antibiotic therapies associated with considerable toxicity. Less toxic and more effective therapies are needed. One promising agent is clofazimine (CFZ), an antibiotic with favorable in vitro data but limited clinical data in RGM. Methods We performed a retrospective cohort study of all patients treated for RGM infection with a CFZ-containing regimen in the University of Pennsylvania Health System between 1/1/2010 and 12/31/2016. Primary outcome was clinical cure, defined as no evidence of clinical or microbiologic infection recurrence after 1 year following the completion of treatment. Secondary outcomes included clinical, radiologic, and microbiologic response; all-cause mortality; infection-specific mortality; and treatment-related adverse events. Descriptive and unadjusted analyses were performed to elucidate associations between pertinent demographic and comorbidity data, clinical presentation, treatment history, and treatment outcomes. Results We treated 55 patients with CFZ for RGM infection during the study period in combination with a median of 5 other antibiotic agents during each treatment course. Clinical cure with initial treatment regimen was achieved in 43% of patients with pulmonary infection and 71% of patients with non-pulmonary infection. CFZ was well tolerated in our cohort and was discontinued prematurely in 20% of patients, but only in the context of discontinuing all antibiotic agents. Conclusions As part of multidrug therapy, CFZ is well tolerated and may be effective in patients with RGM infection, especially non-pulmonary and non-Mycobacterium abscessus complex infections.

Role of epaQ, a Previously Uncharacterized Enterococcus faecalis Gene, in Biofilm Development and Antimicrobial Resistance.

Enterococcus faecalis is a commensal of the human gastrointestinal tract; it is also an opportunistic pathogen and one of the leading causes of hospital-acquired infections. E. faecalis produces biofilms that are highly resistant to antibiotics, and it has been previously reported that certain genes of the epa operon contribute to biofilm-associated antibiotic resistance. Despite several studies examining the epa operon, many gene products of this operon remain annotated as hypothetical proteins. Here, we further explore the epa operon; we identified epaQ, currently annotated as encoding a hypothetical membrane protein, as being important for biofilm formation in the presence of the antibiotic daptomycin. Mutants with disruptions of epaQ were more susceptible to daptomycin relative to the wild type, suggesting its importance in biofilm-associated antibiotic resistance. Furthermore, the ΔepaQ mutant exhibited an altered biofilm architectural arrangement and formed small aggregates in liquid cultures. Our cumulative data show that epa mutations result in altered polysaccharide content, increased cell surface hydrophobicity, and decreased membrane potential. Surprisingly, several epa mutations significantly increased resistance to the antibiotic ceftriaxone, indicating that the way in which the epa operon impacts antibiotic resistance is antibiotic dependent. These results further define the key role of epa in antibiotic resistance in biofilms and in biofilm architecture.IMPORTANCEE. faecalis is a common cause of nosocomial infection, has a high level of antibiotic resistance, and forms robust biofilms. Biofilm formation is associated with increased antibiotic resistance. Therefore, a thorough understanding of biofilm-associated antibiotic resistance is important for combating resistance. Several genes from the epa operon have previously been implicated in biofilm-associated antibiotic resistance, pathogenesis, and competitive fitness in the GI tract, but most genes in this locus remain uncharacterized. Here, we examine epaQ, which has not been characterized functionally. We show that the ΔepaQ mutant exhibits reduced biofilm formation in the presence of daptomycin, altered biofilm architecture, and increased resistance to ceftriaxone, further expanding our understanding of the contribution of this operon to intrinsic enterococcal antibiotic resistance and biofilm growth.

Pan-Genomic and Polymorphic Driven Prediction of Antibiotic Resistance in Elizabethkingia.

The Elizabethkingia are a genetically diverse genus of emerging pathogens that exhibit multidrug resistance to a range of common antibiotics. Two representative species, Elizabethkingia bruuniana and E. meningoseptica, were phenotypically tested to determine minimum inhibitory concentrations (MICs) for five antibiotics. Ultra-long read sequencing with Oxford Nanopore Technologies (ONT) and subsequent de novo assembly produced complete, gapless circular genomes for each strain. Alignment based annotation with Prokka identified 5,480 features in E. bruuniana and 5,203 features in E. meningoseptica, where none of these identified genes or gene combinations corresponded to observed phenotypic resistance values. Pan-genomic analysis, performed with an additional 19 Elizabethkingia strains, identified a core-genome size of 2,658,537 bp, 32 uniquely identifiable intrinsic chromosomal antibiotic resistance core-genes and 77 antibiotic resistance pan-genes. Using core-SNPs and pan-genes in combination with six machine learning (ML) algorithms, binary classification of clindamycin and vancomycin resistance achieved f1 scores of 0.94 and 0.84, respectively. Performance on the more challenging multiclass problem for fusidic acid, rifampin and ciprofloxacin resulted in f1 scores of 0.70, 0.75, and 0.54, respectively. By producing two sets of quality biological predictors, pan-genome genes and core-genome SNPs, from long-read sequence data and applying an ensemble of ML techniques, our results demonstrated that accurate phenotypic inference, at multiple AMR resolutions, can be achieved.

Mycobacterium abscessus Cells Have Altered Antibiotic Tolerance and Surface Glycolipids in Artificial Cystic Fibrosis Sputum Medium.

Mycobacterium abscessus is a biofilm-forming, multidrug-resistant nontuberculous mycobacterial (NTM) pathogen increasingly found in cystic fibrosis patients. Antibiotic treatment for these infections is often unsuccessful, partly due to M. abscessus's high intrinsic antibiotic resistance. It is not clear whether antibiotic tolerance caused by biofilm formation also contributes to poor treatment outcomes. We studied the surface glycolipids and antibiotic tolerance of M. abscessus biofilms grown in artificial cystic fibrosis sputum (ACFS) medium to determine how they are affected by nutrient conditions that mimic infection. We found that M. abscessus displays more of the virulence lipid trehalose dimycolate when grown in ACFS than when grown in standard lab medium. In ACFS medium, biofilm-associated cells were more antibiotic tolerant than planktonic cells in the same well. This contrasts with standard lab media, where both biofilm and planktonic cells are highly antibiotic tolerant. These results indicate that M. abscessus cell physiology in biofilms depends on environmental factors and that nutrient conditions found within cystic fibrosis infections could contribute to both increased virulence and antibiotic tolerance.

Two Pseudomonas aeruginosa clonal groups belonging to the PA14 clade are indigenous to the Churince system in Cuatro Ciénegas Coahuila, México.

Pseudomonas aeruginosa is a widely distributed environmental bacterium but is also an opportunistic pathogen that represents an important health hazard due to its high intrinsic antibiotic resistance and its production of virulence factors. The genetic structure of P. aeruginosa populations using whole genome sequences shows the existence of three clades, one of which (PA7 clade) has a higher genetic diversity. These three clades include clinical and environmental isolates that are very diverse in terms of geographical origins and isolation date. Here, we report the characterization of two distinct clonal P. aeruginosa groups that form a part of the PA14 clade (clade 2) sampled from the Churince system in Cuatro Ciénegas Basin (CCB). One of the clonal groups that we report here was isolated in 2011 (group 2A) and was displaced by the other clonal group (2B) in 2015. Both Churince groups are unable to produce pyoverdine but can produce other virulence-associated traits. The existence of these unique P. aeruginosa clonal groups in the Churince system is of ecological and evolutionary significance since the microbiota of this site is generally very distinct from other lineages, and this is the first time that a population of P. aeruginosa has been found in CCB.

Indole Reverses Intrinsic Antibiotic Resistance by Activating a Novel Dual-Function Importer.

Bacterial antibiotic resistance modulation by small signaling molecules is an emerging mechanism that has been increasingly reported in recent years. Several studies indicate that indole, an interkingdom signaling molecule, increases bacterial antibiotic resistance. However, the mechanism through which indole reduces antibiotic resistance is largely unknown. In this study, we demonstrated a novel mechanism for indole-mediated reversal of intrinsic antibiotic resistance in Lysobacter This reversal was facilitated by a novel BtuD-associated dual-function importer that can transfer both vitamin B12 and antibiotics. Indole stimulated btuD overexpression and promoted efficient absorption of extracellular vitamin B12; meanwhile, the weak selectivity of the importer caused cells to take up excessive doses of antibiotics that resulted in cell death. Consistently, btuD deletion and G48Y/K49D substitution led to marked reductions in the uptake of both antibiotics and vitamin B12 This novel mechanism is common across multiple bacterial species, among which the Q-loop amino acid of BtuD proteins is Glu (E) instead of Gln (Q). Interestingly, the antibiotic resistance of Lysobacter spp. can be restored by another small quorum sensing signaling factor, 13-methyltetradecanoic acid, designated LeDSF, in response to bacterial population density. This work highlights the mechanisms underlying dynamic regulation of bacterial antibiotic resistance by small signaling molecules and suggests that the effectiveness of traditional antibiotics could be increased by coupling them with appropriate signaling molecules.IMPORTANCE Recently, signaling molecules were found to play a role in mediating antibiotic resistance. In this study, we demonstrated that indole reversed the intrinsic antibiotic resistance (IRAR) of multiple bacterial species by promoting the expression of a novel dual-function importer. In addition, population-dependent behavior induced by 13-methyltetradecanoic acid, a quorum sensing signal molecule designated LeDSF, was involved in the IRAR process. This study highlights the dynamic regulation of bacterial antibiotic resistance by small signaling molecules and provides direction for new therapeutic strategies using traditional antibiotics in combination with signaling molecules.

Proanthocyanidin Interferes with Intrinsic Antibiotic Resistance Mechanisms of Gram-Negative Bacteria.

Antibiotic resistance is spreading at an alarming rate among pathogenic bacteria in both medicine and agriculture. Interfering with the intrinsic resistance mechanisms displayed by pathogenic bacteria has the potential to make antibiotics more effective and decrease the spread of acquired antibiotic resistance. Here, it is demonstrated that cranberry proanthocyanidin (cPAC) prevents the evolution of resistance to tetracycline in Escherichia coli and Pseudomonas aeruginosa, rescues antibiotic efficacy against antibiotic‐exposed cells, and represses biofilm formation. It is shown that cPAC has a potentiating effect, both in vitro and in vivo, on a broad range of antibiotic classes against pathogenic E. coli, Proteus mirabilis, and P. aeruginosa. Evidence that cPAC acts by repressing two antibiotic resistance mechanisms, selective membrane permeability and multidrug efflux pumps, is presented. Failure of cPAC to potentiate antibiotics against efflux pump‐defective mutants demonstrates that efflux interference is essential for potentiation. The use of cPAC to potentiate antibiotics and mitigate the development of resistance could improve treatment outcomes and help combat the growing threat of antibiotic resistance.

Antibiotic resistance and heavy metal tolerance in cultured bacteria from hot springs as indicators of environmental intrinsic resistance and tolerance levels

Antibiotic resistance (AR) in the environment is a growing and global concern for public health, and intrinsic AR from pristine sites untouched by pharmaceutical antibiotics is not commonly studied. Forty aerobic bacteria were isolated from water and sediment samples of hot springs in South Africa. Resistance against ten antibiotics (carbenicillin, gentamicin, kanamycin, streptomycin, tetracycline, chloramphenicol, ceftriaxone, co-trimoxazole, nalidixic acid and norfloxacin) was tested using a standard disk diffusion assay. Resistance to one or two antibiotics were equally found in 37.5%, while the remaining 22% showed complete sensitivity. Intermediate resistance was found for ceftriaxone (52.5%), nalidixic acid (37.5%) and carbenicillin (22.5%), while low levels of resistance were observed for streptomycin (5%) and kanamycin (2.5%), and total sensitivity towards the other antibiotics. Twenty-nine isolates were also tested against eight different heavy-metal salts (Al, Cr, Cu, Fe, Hg, Mn, Ni and Pb) at 10 and 40 mM. All isolates were tolerant and able to grow on ≥2 heavy-metal salts at both concentrations. No association was observed between AR and heavy metal tolerance (HMT). Based on the relatively low AR levels, hot spring sites are pristine environments reflecting baseline levels for comparison to other potentially contaminated groundwater sites.

Assessing Food Safety Implications of Multi Antibiotic Resistant Fermented-Food-Condiment-Environment-Adapted Bacteria

Problem of Research: Food safety implications of fermented-condiment-adapted bacterial strains, regarding their intrinsic, acquired and transferable antibiotic resistance potentials are yet to be reportedly fully ascertained.
 Aim: To determine food safety implications of culturalable fermented-condiment-adapted bacterial strains.
 Methodology: Using the Kirby-Bauer agar disc-diffusion method, phenotypic multi-antibiotic-drug-in-discs resistance (MADIDR) profiles of 138 fermented-condiment-borne (Gram-positive = 71; Gram-negative = 67) bacterial strains from iru, ogiri and okpehe were evaluated by in-discs antibiotics, which are commonly administered in human and animal prophylaxis and therapy.
 Results: None of the fermented-condiment-adapted bacterial strains was totally susceptible to the test in-discs antibiotics; just five (3.62%: n = 0.7%: Gram-positive: n = 2.9% Gram-negative) strains were mono-resistant, while six (4.37%) were totally or pandrug-resistant (PDR). Of the remaining 92.03% fermented-condiment-adapted-bacterial strains, 6.57% exhibited co-antibiotic drug resistance (CDR); 43.8% (Gram-positive = 17.52%; Gram-negative = 26.28%) were multi-drug resistant (MDR); and 41.55% (Gram-positive = 25.55%: Gram-negative = 16.0%) displayed extensive-drug resistance (XDR). A total of 43.48% Gram-positive and 36.96% Gram-negative bacterial strains were multi-resistant to between four and eight of the test in-discs antibiotics. Overall, augmentin (95.8%), cloxacillin (94.4%) cotrimoxazole (71.8%) and erythromycin (71.8%) were the most-resisted in-discs antibiotics by the condiment-adapted Gram-positive bacteria, while cloxacillin (93.8%), ciprofloxacin (80.0%) and augmentin (76.9%) were the most-resisted in-discs antibiotics by Gram-negative bacteria.
 Conclusion: Tremendous multi and extensive resistance to in-discs antibiotics were recorded among fermented-food-condiment-environment-adapted bacteria, indicating a serious food safety challenge in the ethnic cottage-food industries, food chain, and the community. Thus, preliminary screening for antibiotic resistance in food-condiment-borne bacteria, using in-discs antibiotics is strongly suggested.