Antimicrobial Multidrug Resistance Research Articles

Liver sinusoidal cells eliminate blood-borne phage K1F.

Phage treatment has regained attention due to an increase in multiresistant bacteria. For phage therapy to be successful, phages must reach their target bacteria in sufficiently high numbers. Blood-borne phages are believed to be captured by macrophages in the liver and spleen. Since liver sinusoids also consist of specialized scavenger liver sinusoidal endothelial cells (LSECs) and Kupffer cells (KCs), this study investigated the contribution of both cell types in the elimination of Escherichia coli phage K1Fg10b::gfp (K1Fgfp) in mice. Circulatory half-life, organ, and hepatocellular distribution of K1Fgfp were determined following intravenous administration. Internalization of K1Fgfp and effects of phage opsonization on uptake were explored using primary mouse and human LSEC and KC cultures. When inoculated with 107 virions, >95% of the total K1Fgfp load was eliminated from the blood within 20 min, and 94% of the total retrieved K1Fgfp was localized to the liver. Higher doses resulted in slower elimination, possibly reflecting temporary saturation of liver scavenging capacity. Phage DNA was detected in both cell types, with a KC:LSEC ratio of 12:1 per population following cell isolation. Opsonization with plasma proteins increased time-dependent cellular uptake in both LSECs and KCs in vitro. Internalized phages were rapidly transported along the endocytic pathway to lysosomal compartments. Reduced viability of intracellular K1Fgfp corroborated inactivation following endocytosis. This study is the first to identify phage distribution in the liver at the hepatocellular level, confirming clearance of K1Fgfp performed mostly by KCs with a significant uptake also in LSECs.IMPORTANCEFaced with the increasing amounts of bacteria with multidrug antimicrobial resistance, phage therapy has regained attention as a possible treatment option. The phage field has recently experienced an emergence in commercial interest as research has identified new and more efficient ways of identifying and matching phages against resistant superbugs. Currently, phages are unapproved drugs in most parts of the world. For phages to reach broad clinical use, they must be shown to be clinically safe and useful. The results presented herein contribute to increased knowledge about the pharmacokinetics of the T7-like phage K1F in the mammalian system. The cell types of the liver that are responsible for rapid phage blood clearance are identified. Our results highlight the need for more research about appropriate dose regimens when phage therapy is delivered intravenously and advise essential knowledge about cell systems that should be investigated further for detailed phage pharmacodynamics.

Antimicrobial resistance and whole genome sequencing of novel sequence types of Enterococcus faecalis, Enterococcus faecium, and Enterococcus durans isolated from livestock

The emergence of antimicrobial-resistant, livestock-associated Enterococcus faecalis represents a public health concern. Here, we report the isolation, molecular detection of virulence and antimicrobial resistance determinants, in addition to the phylogenetic analyses of 20 Enterococcus species using whole genome sequencing analysis of 15 Enterococcus faecalis strains including six strains of three novel sequence types, three Enterococcus faecium and two Enterococcus durans. All strains were isolated from food chain animals in South Africa. Enterococcus strains were isolated on bile aesculin azide agar, followed by identification using MALDI-TOF MS analysis. Antibiotic susceptibility testing was performed using the Kirby–Bauer disk diffusion method. The genomic DNA of the isolates was extracted and sequencing was performed using the Illumina MiSeq platform. Sequence reads were trimmed and de novo assembled. The assembled contigs were analyzed for antimicrobial resistance genes and chromosomal mutations, extra-chromosomal plasmids, and multi-locus sequence type (MLST). Multidrug antimicrobial resistance genes conferring resistance to aminoglycosides (ant(6)-Ia, aph(3′)-IIIa, sat4, and spw), lincosamides (lnu(B), lsa(A), and lsa(E)), macrolides (erm(B)), trimethoprim (dfrG) and tetracyclines (tet(L) and tet(M)) were identified. Plasmid replicons were detected in seven E. faecalis and three E. faecium isolates. The sequence type (ST) of each isolate was determined using the Enterococcus PubMLST database. Ten STs were identified in the collection, three of which (ST1240, ST1241, and ST1242) have not been previously reported and are described in the present study for the first time. To compare the sequenced strains to other previously sequenced E. faecalis strains, assembled sequences of E. faecalis from livestock were downloaded from the PubMLST database. Core genome-based phylogenetic analysis was performed using ParSNP. The detection of multiple drug-resistance in Enterococcus including E. faecalis and E. faecium highlights the significance of genomic surveillance to monitor the spread of antimicrobial resistance in food chain animals. In addition, the genome sequences of Enterococcus strains reported in the present study will serve as a reference point for future molecular epidemiological studies of livestock-associated and antibiotic-resistant E. faecalis in Africa. In addition, this study enables the in-depth analysis of E. faecalis genomic structure, as well as provides valuable information on the phenotypic and genotypic antimicrobial resistance, and the pathogenesis of livestock-associated E. faecalis and E. faecium.

Antimicrobial Multidrug Resistance: Clinical Implications for Infection Management in Critically Ill Patients.

The increasing incidence of antimicrobial resistance (AMR) worldwide represents a serious threat in the management of sepsis. Due to resistance to the most common antimicrobials prescribed, multidrug-resistant (MDR) pathogens have been associated with delays in adequate antimicrobial therapy leading to significant increases in mortality, along with prolonged hospital length of stay (LOS) and increases in healthcare costs. In response to MDR infections and the delay of microbiological results, broad-spectrum antibiotics are frequently used in empirical antimicrobial therapy. This can contribute to the overuse and misuse of antibiotics, further promoting the development of resistance. Multiple measures have been suggested to combat AMR. This review will focus on describing the epidemiology and trends concerning MDR pathogens. Additionally, it will explore the crucial aspects of identifying patients susceptible to MDR infections and optimizing antimicrobial drug dosing, which are both pivotal considerations in the fight against AMR. Expert commentary: The increasing AMR in ICUs worldwide makes the empirical antibiotic therapy challenging in septic patients. An AMR surveillance program together with improvements in MDR identification based on patient risk stratification and molecular rapid diagnostic tools may further help tailoring antimicrobial therapies and avoid unnecessary broad-spectrum antibiotics. Continuous infusions of antibiotics, therapeutic drug monitoring (TDM)-based dosing regimens and combination therapy may contribute to optimizing antimicrobial therapy and limiting the emergence of resistance.

Photodynamic Therapy Applications in Cancer, Infections and Cardiovascular Disease

Photodynamic therapy (PDT) or light treatment is a procedure to treat certain types of diseases that uses a photosensitizer (PS) which upon light activation produces cytotoxic oxygen species destroying tumor cells. PDT is minimally-invasive and can be repeated a few times without accumulating significant toxicity in the surrounding tissues. While PDT application in cancer treatment is well established and widely recognized by the scientific community, and its use in the fight against microbial infection is gaining momentum with the rapid increase in antimicrobial multi-drug resistance, its use in the treatment of cardiovascular diseases is clearly marginal. This mini-review presents a brief historical background of PDT, photo-activatable agents utilized, mechanism of photodynamic action, studies in vitro and in vivo, clinical trials, limitations and future prospects of its application in various types of cancer, infections caused by pathogens such as COVID-19, and heart diseases.

ASPECTS OF MUSHROOMS AND THEIR EXTRACTS AS NATURAL ANTIMICROBIAL AGENTS

Nowadays, multidrug antimicrobial resistance is a very common issue globally. Antimicrobial resistance is fueling the fire, and it has been noted that infectious disease incidence and deaths are rising throughout the world. Many pathogens have been reported to develop resistance to these synthetic and semi-synthetic drugs leading to minimized efficacy and resulting in substantial economic losses all over the world. A new approach for choosing natural extracts of plants and fungi as medicines and natural antibacterial agents has been developed to address this challenge. According to recent investigations and research, mushrooms and their extracts contain bioactive compounds that can be used as a natural antibacterial alternative against a number of bacterial species. In addition to having medical properties like antibacterial, anticancer, and antioxidant activity, mushrooms are a highly nutritious food source. The present study emphasized on the antimicrobial resistance, mechanisms and antibacterial properties of mushrooms.

Prevalent Dominant Acinetobacter baumannii ST191/195/208 Strains in Bloodstream Infections Have High Drug Resistance and Mortality.

Different sequence types of Acinetobacter baumannii (AB) have their own epidemiological characteristics, drug resistance, and toxicity. AB bloodstream infection (BSI) in the First Affiliated Hospital of Medical College of Zhejiang University from January 2012 to December 2017 were classified by multilocus sequence typing. Clinical data of patients were retrospectively analyzed, drug resistance and toxicity were respectively studied by drug sensitivity and complement killing tests. 247 unduplicated AB strains were collected, and ST191/195/208, the main epidemic dominant strain, accounted for 70.9%. Patients with ST191/195/208 on infection had higher white blood cell (10.8 vs 8.9, p = 0.004), neutrophil% (89.5 vs 86.9, p = 0.005), neutrophil count (9.5 vs 7.1, p = 0.021), D-dimer (6.7 vs 3.8, p = 0.000), total bilirubin (27.0 vs 21.5, p = 0.038), pronatriuretic peptide (324 vs 164, p = 0.042), C-reactive protein (82.5 vs 56.3, p = 0.048), clinical pulmonary infection score (CPIS; 7.33 ± 2.30 vs 6.50 ± 2.72, p = 0.045), and acute physiology and chronic health evaluation-II (APACHE-II; 19.620 ± 5.1850 vs 17.648 ± 6.1251, p = 0.011). Patients with ST191/195/208 had more complications, including pulmonary infection (p = 0.041), septic shock (p = 0.009), and multiple organ failure (p = 0.019). Patients with ST191/195/208 had higher 3 day mortality (24.6% vs 13.9%, p = 0.043), 14 day mortality (46.8% vs 26.8%, p = 0.003), and 28 day mortality (55.0% vs 32.4%, p = 0.001). ST191/195/208 strains had higher drug resistance to most antibiotics, and higher survival rate at 90% normal serum concentration (p < 0.001). ST191/195/208 strains predominate in the hospital and prevails in patients with severe infections with increased multidrug antimicrobial resistance and excessive mortality compared to any other AB stains.

A novel T4- and λ-based receptor binding protein family for bacteriophage therapy host range engineering.

Widespread multidrug antimicrobial resistance in emerging pathogens has led to a renewed interest in phage therapy as an alternative or supplement to traditional small molecule drugs. The primary limiting factors of phage therapy deployment rest in the narrow host range specificity of phage as well as a poor understanding of many phages' unintended downstream effects on host physiology and microbiota as well as on adverse pathogen evolution. Consequently, this has made assembling well-defined and safe "phage-cocktails" of solely naturally occurring phages labor- and time-intensive. To increase the speed, efficacy, and safety of therapeutic deployment, there is exceptional interest in modulating the host ranges of well-characterized lytic phages (e.g., T4 and T7) by using synthetic strategies to the swap phage tail components, the receptor binding proteins (RBPs) key for host specificity. Here we identify the RBP of the Citrobacter rodentium temperate phage ΦNP as ORF6. Through bioinformatic and phylogenetic assays, we demonstrate this RBP to be closely related to the known RBPs of T4 and λ. Further investigation reveals a novel, greater than 200 members RBP family with phages targeting several notable human pathogens, including Klebsiella pneumoniae, Escherichia coli O157:H7, Salmonella spp., and Shigella spp. With well characterized lytic members, this RBP family represents an ideal candidate for use in synthetic strategies for expanding therapeutic phage host ranges.

Surveillance for multidrug resistant Escherichia coli carriage in cattle, dogs and humans reveals predominance of CMY-2, CTX-M-15 and CTX-M-9 groups of β-lactamases

Global spread of antimicrobial multidrug resistance (MDR) in human and veterinary medicine relies upon diagnostics, surveillance and stewardship to guide mitigation. Utilizing surveillance of fecal samples from our service area for detecting MDR Escherichia coli carriage in humans (2143), dogs (627), and cattle (130), we found isolates resistant to third/fourth generation cephems present in 3.7 %, 13.1 %, and 51.5 %, respectively. CMY-2, CTX-M-15-like and CTX-M9 group genes in descending order were predominant in all hosts and accounted for 83.3 % of non-wild-type gene targets. MDR carriage mirrored cephem non-susceptibility rates as published in annual antibiograms for humans and dogs; notably, no carbapenem-resistant carriage isolates were detected. Given the scale of MDR E. coli carriage in cattle (14X) and dogs (3.5X) compared to humans, bench-marking of the resistance gene pool by host species utilizing regional One Health surveillance may aid in assessing occupational and geographic risks for acquiring resistance and for monitoring of mitigation strategies.

Draft Genome Sequence of Multidrug Resistant Klebsiella pneumoniae Strain C43 Isolated from Environmental Water Sample.

ABSTRACTHere, we report the genome of ESBL-producing Klebsiella pneumoniae strain C43, which was isolated from an environmental water sample. The genome is 5,614,412 bp in size with GC content of 56.86% with multidrug antimicrobial resistance genes and several metal resistance gene operons.

Antibacterial, Antibiofilm, and Antiviral Farnesol-Containing Nanoparticles Prevent Staphylococcus aureus from Drug Resistance Development.

Multidrug antimicrobial resistance is a constantly growing health care issue associated with increased mortality and morbidity, and huge financial burden. Bacteria frequently form biofilm communities responsible for numerous persistent infections resistant to conventional antibiotics. Herein, novel nanoparticles (NPs) loaded with the natural bactericide farnesol (FSL NPs) are generated using high-intensity ultrasound. The nanoformulation of farnesol improved its antibacterial properties and demonstrated complete eradication of Staphylococcus aureus within less than 3 h, without inducing resistance development, and was able to 100% inhibit the establishment of a drug-resistant S. aureus biofilm. These antibiotic-free nano-antimicrobials also reduced the mature biofilm at a very low concentration of the active agent. In addition to the outstanding antibacterial properties, the engineered nano-entities demonstrated strong antiviral properties and inhibited the spike proteins of SARS-CoV-2 by up to 83%. The novel FSL NPs did not cause skin tissue irritation and did not induce the secretion of anti-inflammatory cytokines in a 3D skin tissue model. These results support the potential of these bio-based nano-actives to replace the existing antibiotics and they may be used for the development of topical pharmaceutic products for controlling microbial skin infections, without inducing resistance development.

Type Strains of Entomopathogenic Nematode-Symbiotic Bacterium Species, Xenorhabdus szentirmaii (EMC) and X. budapestensis (EMA), Are Exceptional Sources of Non-Ribosomal Templated, Large-Target-Spectral, Thermotolerant-Antimicrobial Peptides (by Both), and Iodinin (by EMC).

Antimicrobial multidrug resistance (MDR) is a global challenge, not only for public health, but also for sustainable agriculture. Antibiotics used in humans should be ruled out for use in veterinary or agricultural settings. Applying antimicrobial peptide (AMP) molecules, produced by soil-born organisms for protecting (soil-born) plants, seems a preferable alternative. The natural role of peptide-antimicrobials, produced by the prokaryotic partner of entomopathogenic-nematode/bacterium (EPN/EPB) symbiotic associations, is to sustain monoxenic conditions for the EPB in the gut of the semi-anabiotic infective dauer juvenile (IJ) EPN. They keep pathobiome conditions balanced for the EPN/EPB complex in polyxenic (soil, vanquished insect cadaver) niches. Xenorhabdus szentirmaii DSM16338(T) (EMC), and X. budapestensis DSM16342(T) (EMA), are the respective natural symbionts of EPN species Steinernema rarum and S. bicornutum. We identified and characterized both of these 15 years ago. The functional annotation of the draft genome of EMC revealed 71 genes encoding non-ribosomal peptide synthases, and polyketide synthases. The large spatial Xenorhabdus AMP (fabclavine), was discovered in EMA, and its biosynthetic pathway in EMC. The AMPs produced by EMA and EMC are promising candidates for controlling MDR prokaryotic and eukaryotic pathogens (bacteria, oomycetes, fungi, protozoa). EMC releases large quantity of iodinin (1,6-dihydroxyphenazine 5,10-dioxide) in a water-soluble form into the media, where it condenses to form spectacular water-insoluble, macroscopic crystals. This review evaluates the scientific impact of international research on EMA and EMC.

Interpretable clinical time-series modeling with intelligent feature selection for early prediction of antimicrobial multidrug resistance

Electronic health records provide rich, heterogeneous data about the evolution of the patients’ health status. However, such data need to be processed carefully, with the aim of extracting meaningful information for clinical decision support. In this paper, we leverage interpretable (deep) learning and signal processing tools to deal with multivariate time-series data collected from the Intensive Care Unit (ICU) of the University Hospital of Fuenlabrada (Madrid, Spain). The presence of antimicrobial multidrug-resistant (AMR) bacteria is one of the greatest threats to the health system in general and to the ICUs in particular due to the critical health status of the patients therein. Thus, early identification of bacteria at the ICU and early prediction of their antibiotic resistance are key for the patients’ prognosis. While intelligent data-based processing and learning schemes can contribute to this early prediction, their acceptance and deployment in the ICUs require the automatic schemes to be not only accurate but also understandable by clinicians. Accordingly, we have designed trustworthy intelligent models for the early prediction of AMR based on the combination of meaningful feature selection with interpretable recurrent neural networks. These models were created using irregularly sampled clinical measurements, both considering the health status of the patient and the global ICU environment. We explored several strategies to cope with strongly imbalance data, since only a few ICU patients are infected by AMR bacteria. It is worth noting that our approach exhibits a good balance between performance and interpretability, especially when considering the difficulty of the classification task at hand. A multitude of factors are involved in the emergence of AMR (several of them not fully understood), and the records only contain a subset of them. In addition, the limited number of patients, the imbalance between classes, and the irregularity of the data render the problem harder to solve. Our models are also enriched with SHAP post-hoc interpretability and validated by clinicians who considered model understandability and trustworthiness of paramount concern for pragmatic purposes. Moreover, we use linguistic fuzzy systems to provide clinicians with explanations in natural language. Such explanations are automatically generated from a pool of interpretable rules that describe the interaction among the most relevant features identified by SHAP. Notice that clinicians were especially satisfied with new insights provided by our models. Such insights helped them to trust the automatic schemes and use them to make (better) decisions to mitigate AMR spreading in the ICU. All in all, this work paves the way towards more comprehensible time-series analysis in the context of early AMR prediction in ICUs and reduces the time of detection of infectious diseases, opening the door to better hospital care.

Photodynamic Inactivation of Antibiotic-Resistant and Sensitive Aeromonas hydrophila with Peripheral Pd(II)- vs. Zn(II)-Phthalocyanines.

The antimicrobial multidrug resistance (AMR) of pathogenic bacteria towards currently used antibiotics has a remarkable impact on the quality and prolongation of human lives. An effective strategy to fight AMR is the method PhotoDynamic Therapy (PDT). PDT is based on a joint action of a photosensitizer, oxygen, and light within a specific spectrum. This results in the generation of singlet oxygen and other reactive oxygen species that can inactivate the pathogenic cells without further regrowth. This study presents the efficacy of a new Pd(II)- versus Zn(II)-phthalocyanine complexes with peripheral positions of methylpyridiloxy substitution groups (pPdPc and ZnPcMe) towards Gram-negative bacteria Aeromonas hydrophila (A. hydrophila). Zn(II)-phthalocyanine, ZnPcMe was used as a reference compound for in vitro studies, bacause it is well-known with a high photodynamic inactivation ability for different pathogenic microorganisms. The studied new isolates of A. hydrophila were antibiotic-resistant (R) and sensitive (S) strains. The photoinactivation results showed a full effect with 8 µM pPdPc for S strain and with 5 µM ZnPcMe for both R and S strains. Comparison between both new isolates of A. hydrophila (S and R) suggests that the uptakes and more likely photoinactivation efficacy of the applied phthalocyanines are independent of the drug sensitivity of the studied strains.

Design, molecular docking and antimicrobial assessment of newly synthesized p-cuminal-sulfonamide Schiff base derivatives

Antibiotic resistance of bacteria and fungi has been brewing for decades and has now surfaced as a potential public health emergency, globally. Thus, newer potent drug candidates are needed urgently to overcome antibiotic-resistant episodes. As an attempt to overcome the antimicrobial multi-drug resistance problems, a new series of p-cuminal sulfonamide Schiff base congeners 3a-3h were designed, synthesized, and their structures confirmed by several spectral studies. The antimicrobial activities assessment of obtained products was carried out against pathogenic bacteria Staphylococcus aureus, Streptococcus pyogenes, Methicillin-resistant Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and against pathogenic fungi Candida tropicalis and Trichophyton rubrum. Among them, 3b exhibited significant inhibition of all the strains of bacteria with MIC as 3.12μg/mL and the compound 3e exhibited the most promising control against both C. tropicalis and T. rubrum (MIC 6.25μg/mL) in comparison to Ketoconazole as well as good inhibition against both S. aureus and MRSA at MIC 6.25 μg/mL The presence of sulfamoyl and azomethine groups and some of heteroaryl rings in individual molecules could have increased the antibacterial actions, while the antifungal action could be attributed to the presence of guanyl and acetyl groups in the molecular structure. Furthermore, in silico investigation and drug-likeness have been ascertained with biological targets.

Molecular characterization of netB and tpeL virulence factors and antimicrobial resistance genes of Clostridium perfringens isolated from herbs and spices.

The present study aimed to determine some virulence-associated genes and antimicrobial multidrug resistance of Clostridium perfringens recovered from herbs and spices widely distributed in the Egyptian market. C. perfringens virulence and resistance factors were determined using PCR targeting the netB, tpeL, ermB, bla and tetK genes. Thirty three out of 392 samples (8.42%) from herbs and spices submitted to our laboratory for bacteriological screening were positive for presence C. perfringens. PCR results for the tpeL gene in isolated C. perfringens revealed 9 out of 33 (27.3 %) of isolates, while netB was not detected. The isolates were resistant to Clindamycin, Vancomycin, tetracycline, and erythromycin with inhibition zones of 6.28 ± 0.63, 8.78 ± 0.41, 9.63 ± 0.63, and 9.84 ± 0.66 mm, respectively. The genes mentioned above were selected to correspond to the ineffective antimicrobials; ermB for erythromycin, tetK for tetracycline, and bla for the remainder. PCR results for antibacterial resistant genes in isolated C. perfringens revealed their presence. From 33 isolates, bla gene was detected in 21 (63.4 %), tetK in 13 (39.4 %) and ermB in only one isolate (3.03 %). Sequencing analysis was done for the bla gene as an example for the detected genes as detected at the highest incidence (63.4%). No cross-relationship was detected upon comparing incidence data of both studied virulence genes and those of antimicrobial resistance. The present findings may explain the resistance of C. perfringens to the examined antibacterials and recommend avoiding the application of them to control the microbe. In addition, the authors recommend following strict hygienic procedures during the industry of herbs and spices to ensure their clearance from Clostridium perfringens before distributing the products as food additives into the markets.

Draft Genome Sequence of Enterobacter hormaechei subsp. steigerwaltii Strain BEI01

ABSTRACTHere, we report the genome sequence of Enterobacter hormaechei subsp. steigerwaltii strain BEI01, originally deposited as a member of the Enterobacter cloacae complex. The genome is 4,900,246 bp in size with a GC content of 55.44%; it contains multidrug antimicrobial resistance genes and several metal resistance gene operons.

Comparative genomics of ST5 and ST30 methicillin-resistant Staphylococcus aureus sequential isolates recovered from paediatric patients with cystic fibrosis

Staphylococcus aureus chronic airway infection in patients with cystic fibrosis (CF) allows this pathogen to adapt over time in response to different selection pressures. We have previously shown that the main sequence types related to community-acquired methicillin-resistant S. aureus (MRSA) infections in Argentina – ST5 and ST30 – are also frequently isolated from the sputum of patients with CF, but in these patients they usually display multi-drug antimicrobial resistance. In this study, we sequenced the genomes of MRSA from four paediatric CF patients with the goal of identifying mutations among sequential isolates, especially those possibly related to antimicrobial resistance and virulence, which might contribute to the adaptation of the pathogen in the airways of patients with CF. Our results revealed genetic differences in sequential MRSA strains isolated from patients with CF in both their core and accessory genomes. Although the genetic adaptation of S. aureus was distinct in different hosts, we detected independent mutations in thyA, htrA, rpsJ and gyrA – which are known to have crucial roles in S. aureus virulence and antimicrobial resistance – in isolates recovered from multiple patients. Moreover, we identified allelic variants that were detected in all of the isolates recovered after a certain time point; these non-synonymous mutations were in genes associated with antimicrobial resistance, virulence, iron scavenging and oxidative stress resistance. In conclusion, our results provide evidence of genetic variability among sequential MRSA isolates that could be implicated in the adaptation of these strains during chronic CF airway infection.

Multidrug Antimicrobial Resistance and Molecular Detection of mcr-1 Gene in Salmonella Species Isolated from Chicken.

Simple SummaryColistin is a widely used antibiotic against infections caused by extensively drug-resistant Gram-negative bacteria. It is critical to track and monitor the presence of mcr-like genes and colistin resistance to protect a last resort treatment against highly antibiotic-resistant bacteria. In the present study, the colistin resistance gene mcr-1 was investigated, and its in-silico functional analysis in Salmonella isolates was determined. Out of 100 chicken samples (liver and intestine), 82 Salmonella spp. were isolated and characterized. Antimicrobial sensitivity was determined using different antimicrobial agents. The isolates were characterized using PCR targeting genus-specific invA and mcr-1 genes followed by in-silico functional analysis. The majority of isolates (92.68%) was found highly resistant to colistin, which demonstrated the occurrence of the colistin resistance mcr-1 gene in Salmonella isolates of chicken origin in Bangladesh. The study also showed the in-silico functional analysis and phylogenetic relationship of the colistin resistance mcr-1 gene among Salmonella isolates. The findings of the present study highlight the increasing issue of transferable colistin resistance and call for immediate action and measures to review the imprudent use of colistin in poultry production systems in Bangladesh.Colistin (polymyxin E) is widely used in animal and human medicine and is increasingly used as one of the last-resort antibiotics against Gram-negative bacilli. Due to the increased use of colistin in treating infections caused by multidrug-resistant Gram-negative bacteria, resistance to this antibiotic ought to be monitored. The study was undertaken to elucidate the molecular mechanisms, genetic relationships and phenotype correlations of colistin-resistant isolates. Here, we report the detection of the mcr-1 gene in chicken-associated Salmonella isolates in Bangladesh and its in-silico functional analysis. Out of 100 samples, 82 Salmonella spp. were isolated from chicken specimens (liver, intestine). Phenotypic disc diffusion and minimum inhibitory concentration (MIC) assay using different antimicrobial agents were performed. Salmonella isolates were characterized using PCR methods targeting genus-specific invA and mcr-1 genes with validation for the functional analysis. The majority of the tested Salmonella isolates were found resistant to colistin (92.68%), ciprofloxacin (73.17%), tigecycline (62.20%) and trimethoprim/sulfamethoxazole (60.98%). When screened using PCR, five out of ten Salmonella isolates were found to carry the mcr-1 gene. One isolate was confirmed for Salmonella enterica subsp. enterica serovar Enteritidis, and other four isolates were confirmed for Salmonella enterica subsp. enterica serovar Typhimurium. Sequencing and phylogenetic analysis revealed a divergent evolutionary relationship between the catalytic domain of Neisseria meningitidis lipooligosaccharide phosphoethanolamine transferase A (LptA) and MCR proteins, rendering them resistant to colistin. Three-dimensional homology structural analysis of MCR-1 proteins and molecular docking interactions suggested that MCR-1 and LptA share a similar substrate binding cavity, which could be validated for the functional analysis. The comprehensive molecular and in-silico analyses of the colistin resistance mcr-1 gene of Salmonella spp. of chicken origin in the present study highlight the importance of continued monitoring and surveillance for antimicrobial resistance among pathogens in food chain animals.

Antimicrobial activity of polypyrrole nanoparticles and aqueous extract of Moringa oleifera against Staphylococcus spp. carriers of multi-drug efflux system genes isolated from dairy farms.

Our objectives were to identify genes of the multi-drug efflux system and to evaluate the antimicrobial activities of polypyrrole nanoparticles (PPy-NPs) and aqueous extract of Moringa oleifera against Staphylococcus spp. isolated from dairy farms in Northeast Brazil. Initially, 162 Staphylococcus spp. isolates were subjected to in vitro antimicrobial sensitivity tests. Of these, 35 presented antimicrobial multi-drug resistance phenotypes. These 35 isolates were then referred for the detection of norA, norB, norC, msrA, mgrA, tet-38, and lmrS genes, all of which feature in multi-drug efflux systems. In the isolates carrying the genes, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PPy-NPs and Moringa oleifera aqueous extract were determined. In the molecular analysis of the 35 isolates norA, norC, tet-38, and msrA genes were detected and for the other genes norB, lmrS and mgrA there was no amplification. Antimicrobial activity was verified of PPy-NPs and aqueous extract of Moringa oleifera in Staphylococcus spp. carrying multi-drug efflux system genes. We concluded that there are multi-drug efflux system genes present in the Staphylococcus spp. from the agricultural environment in Northeast Brazil, and that aqueous extract of Moringa oleifera and PPy-NPs show bactericidal activity against these isolates.

Complex dynamical behavior of a discretize fractional-order multi-drug antimicrobial resistance model

In this paper, we proposed and investigated a discretized form of a fractional-order multi-drug antimicrobial resistance model. The existence and the uniqueness of the proposed model is studied. The impacts of varying the parameters of the system are examined. Under certain conditions, the system undergoes some kinds of bifurcations. It is established that the fractional-order parameter affects the stability of the model. The model has an opulent diversity of dynamical behaviors such as Hopf bifurcation, chaotic attractors and attractor crisis. We used some numerical simulations to illustrate our analytical results.