Multi-antibiotic Resistant Bacteria Research Articles

A scientific research training programme for teaching biomedical students to identify the horizontal transfer of antibiotic resistance genes.

Worldwide prevalence of multi-antibiotic resistant bacteria is rapidly increasing, and the education of undergraduates and graduates about antibiotic resistance and its associated horizontal gene transfer is critical in the general effort to confront the spread of antibiotic resistance. In this study, a deeper understanding of antibiotic resistance and horizontal gene transfer was achieved by biomedical undergraduate students through a scientific research programme. The enthusiasm of students to participate in the training programme was very high, and results revealed that each student could identify the antibiotic resistance integrative and conjugative element from the Stenotrophomonas maltophilia MER1 genome. Each student could also draw the phylogenetic relationship of the antibiotic resistance integrative and conjugative element. In addition, students proved the horizontal transfer of antibiotic resistance genes from S. maltophilia MER1 to Escherichia coli strain 25DN through conjugation and PCR assays. Each group of students was able to obtain the expected results, indicating that the outcome of the scientific research programme was highly reproducible. This programme improved the theoretical knowledge about antibiotic resistance and horizontal gene transfer and the research skills of biomedical sciences students. Through this programme, students learned that antibiotic resistance genes can be horizontally transferred among different bacteria, laying a solid foundation for students to value the importance of the appropriate use of antibiotics in their future work and life.

Using UV/peracetic acid as pretreatment for subsequent bio-treatment of antibiotic-containing wastewater treatment: Mitigating microbial inhibition and antibiotic resistance genes proliferation

UV/peracetic acid (PAA) treatment presents a promising approach for antibiotic removal, but its effects on microbial community and proliferation of antibiotic resistance genes (ARGs) during the subsequent bio-treatment remain unclear. Thus, we evaluated the effects of the UV/PAA on tetracycline (TTC) degradation, followed by introduction of the treated wastewater into the bio-treatment system to monitor changes in ARG expression and biodegradability. Results demonstrated effective TTC elimination by the UV/PAA system, with carbon-centered radicals playing a significant role. Crucially, the UV/PAA system not only eliminated antibacterial activity but also inhibited potential ARG host growth, thereby minimizing the emergence and dissemination of ARGs during subsequent bio-treatment. Additionally, the UV/PAA system efficiently removed multi-antibiotic resistant bacteria and ARGs from the bio-treatment effluent, preventing ARGs from being released into the environment. Hence, we propose a multi-barrier strategy for treating antibiotic-containing wastewater, integrating UV/PAA pre-treatment and post-disinfection with bio-treatment. The inhibition of ARGs transmission by the integrated system was verified through actual soil testing, confirming its effectiveness in preventing ARGs dissemination in the surrounding natural ecosystem. Overall, the UV/PAA treatment system offers a promising solution for tackling ARGs challenges by controlling ARGs proliferation at the source and minimizing their release at the end of the treatment process.

Detection of genes conferring resistance on multi antibiotic resistance bacteria isolated from ready to eat foods sold in the Nigerian defence academy, Kaduna, Nigeria

Food has always been the means by which disease-causing organisms are transmitted to man. This study sought to isolate multi-antibiotic resistant bacteria from ready to eat food (RTE) sold within Nigerian Defence Academy. Eight (8) food samples were collected from five (5) vendors and spread plate technique was adopted for bacteria isolation from the food samples. Twenty-five (25) bacteria isolates were obtained from the food samples and subjected to antibiotic sensitivity assay using Kirby Bauer disc diffusion method. Eight (8) of the isolates exhibited various degrees of resistance to the tested antibiotics. All the isolated bacteria in this study were susceptible to the effect of Quinolones antibiotics and were resistant to cefuroxime, cefotaxime and Ampiclox. Six (6) of the isolates that showed multi-antibiotic resistance were characterized by 16SrRNA gene sequencing as Bacillus cereus strains (0083 and HYM76), Staphylococcus aureus strain MRSA252, Lactobacillus murinus strain AMB_10 and Listeria Spp. Polymerase chain reaction (PCR) and agarose gel electrophoresis detection of four (4) antibiotics resistance genes – Aminoglycoside adenylyltransferase (AADA1), Acridine (ACRA), Chloramphenicol Resistance (CMLA) and Erythromycin (ERM(C)) – produced the following results: AADA1 genes were amplified in Staphylococcus aureus and Bacillus cereus; ACRA gene was detected in all the selected isolates; CMLA gene was detected in Staphylococcus aureus, Lactobacillus murinus and Bacillus cereus while ERM(C) gene was present in Staphylococcus aureus.

A three-in-one nanoplatform with photo/chemodynamic activities combined with glutathione depletion for treating bacterial infections

The misuse of antibiotics leading to bacterial multidrug resistance is responsible for severe infectious diseases and a significant cause of mortality worldwide, resulting in numerous human disasters. Photodynamic antibacterial therapy (PDAT) is a promising strategy against multiantibiotic-resistant bacteria, but its antibacterial activity is greatly limited by reduced glutathione (GSH) in bacteria. In this study, we constructed a nanoplatform through the formation of metal chelating complexes (FeP) between ferric and pyrophosphate ions, with subsequent adsorption of the photosensitizer ZnPc(COOH)8 (octa-carboxyl substituted zinc phthalocyanine) mediated by polylysine (PL) on the surface. The nanocomplexes FeP@PL:ZnPc(COOH)8 exhibited the capacity of GSH depletion and chemodynamic activity, which synergistically enhanced PDAT efficacy. FeP@PL:ZnPc(COOH)8 possessed the excellent antibacterial activity in vivo and in vitro, which might be attributed to the increased production of reactive oxygen species (ROS), reduced GSH level in bacteria, improved bacterial uptake, and enhanced destruction of the bacterial outer membrane. Moreover, FeP@PL:ZnPc(COOH)8 exhibited accelerated wound healing efficacy and the ability to recognize bacteria-infected wounds, rendering it an effective theranostic nanoplatform for bacterial infections. The construction strategy of nanocomplexes in this study holds theoretical and practical significance for high-efficiency synergistic photodynamic and chemodynamic antibacterial therapy.

Illustration on phenotypic and genotypic characteristics of typical multi-antibiotic resistant bacteria in aquatic environments through complete genomes and comparative genomics

Antibiotic-resistant bacteria, especially multi-antibiotic-resistant bacteria (MARBs), greatly threaten environmental safety and human health. However, studies on the phenotypic resistance and complete genotypic characterization of MARB in aquatic environments are lacking. In this study, a multi-resistant superbug (TR3) was screened by the selective pressure of multi-antibiotics from the activated sludge of the aeration tanks of urban wastewater treatment plants (WWTPs) in 5 different regions of China. Based on the 16 S rDNA sequence alignment it was found that the sequence similarity between strain TR3 and Aeromonas was as high as 99.50%. The genome-wide sequence showed that the base content of the chromosome of strain TR3 is 4,521,851 bp. It contains a plasmid with a length of 9182 bp. All antibiotic resistance genes (ARGs) of strain TR3 are located on the chromosome, which means that it has passage stability. There are multiple types of resistance genes in the genome and plasmid of strain TR3, enduing it with resistance to 5 antibiotics (ciprofloxacin, tetracycline, ampicillin, clarithromycin, and kanamycin), accompanied by the strongest resistance to kanamycin (aminoglycosides) and the worst resistance to clarithromycin (quinolones). From the perspective of gene expression, we show the resistance mechanism of strain TR3 to different types of antibiotics. In addition, the potential pathogenicity of strain TR3 is also discussed. Chlorine and ultraviolet (UV) sterilization on strain TR3 showed that UV is ineffective at low intensity, and it is easy to be revived by light. A low concentration of hypochlorous acid is effective for sterilization, but it can cause the release of DNA, becoming a potential source of ARGs discharged from WWTPs to environmental water bodies.

Promising antibacterial activities of anethole and green-synthesized magnetite nanoparticles against multiple antibiotic-resistant bacteria.

The emergence of antibiotic resistance is considered a major threat, and this problem is exacerbated due to the excessive use of antibiotics. Therefore, it is necessary to find new antimicrobials that are effective against pathogenic bacteria resistant to existing antimicrobials. This study presents a novel antimicrobial approach for the bio-control of multi-antibiotic-resistant (MAR) bacteria. Ten antibiotic discs were used to evaluate the bacterial sensitivity. Five bacterial strains showed resistance to all the tested antibiotic groups. The following MAR bacterial strains were selected, and their identification was confirmed by 16S rRNA with an accession number on the NCBI: Shigella flexneri MZ350855.1; Klebsiella aerogenes MZ352107.1; Serratia marcescens MZ618709.1; Yersinia enterocolitica MZ673567.1; and Achromobacter denitrificans OK560350.1. Star anise extract (SAE) gives the highest inhibitory efficiency against isolated MAR bacteria between three local plant extracts. SAE was used to synthesize magnetite nanoparticles (MNPs) using a simple and eco-friendly green biosynthesis technique. Synthesized MNPs were characterized by X-ray diffraction, ultraviolet-visible spectroscopy, Fourier transform infrared, and transmission electron microscopy techniques. The effect of synthesized magnetic nanoparticles for wastewater treatment in bacterial portion demonstrated remarkable bactericidal performance against total bacterial count with a percentage reduction of 84% using 0.05 g, and the efficiency of decreasing heavy metals was estimated.

Effect of Agrimonia eupatoria L. and Origanum vulgare L. Leaf, Flower, Stem, and Root Extracts on the Survival of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is one of the most antibiotic multi-resistant bacteria, causing chronic pulmonary disease and leading to respiratory failure and even mortality. Thus, there has been an ever-increasing search for novel and preferably natural antimicrobial compounds. Agrimonia eupatoria L. and Origanum vulgare L. shoots are commonly used as teas or alcoholic tinctures for their human health-promoting and antibacterial properties. Here, we explored the antimicrobial effects of all plant parts, i.e., leaf, flower, stem, and root extracts, prepared in water or in 60% ethanol, against P. aeruginosa. The impact of these extracts on bacterial survival was determined using a luminescent strain of P. aeruginosa, which emits light when alive. In addition, the antimicrobial effects were compared with the antioxidant properties and content of phenolic compounds of plant extracts. Ethanolic extracts of O. vulgare roots and flowers showed the highest antimicrobial activity, followed by A. eupatoria roots. In particular, chlorogenic acid, the ethanolic extract of O. vulgare roots contained high levels of protocatechuic acid, hesperidin, shikimic acid, rutin, quercetin, and morin. The synergistic effects of these phenolic compounds and flavonoids may play a key role in the antibacterial activity of teas and tinctures.

LL-37-Coupled Porous Composite Scaffold for the Treatment of Infected Segmental Bone Defect.

Increased multiantibiotic-resistant bacteria means that infected bone defects remain a significant challenge to clinics. Great interest has emerged in the use of non-antibiotic antimicrobials to reduce the rate of multiantibiotic-resistant bacterial infection and facilitate bone regeneration. The cationic antimicrobial peptide LL-37 is the sole human cathelicidin and has shown nonspecific activity against a broad spectrum of microorganisms. In this study, we fabricated the poly(lactic-co-glycolic acid)/β-calcium phosphate/peptide LL-37 (PLGA/TCP/LL-37, PTL) scaffold with low-temperature 3D-printing technology for the treatment of infected segmental bone defects. The prepared scaffolds were divided into three groups: a high LL-37 concentration group (PTHL), low LL-37 concentration group (PTLL) and blank control group (PT). The cytocompatibility and antimicrobial activity of the engineered scaffolds were tested in vitro, and their osteogenesis properties were assessed in vivo in a rat infected bone defect model. We found the fabricated PTL scaffold had a well-designed porous structure that could support a steady and prolonged LL-37 release. Furthermore, the PTHL group showed strong antibacterial activity against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) without any inhibition of the proliferation or alkaline phosphatase activity of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. In addition, the infected femoral defects implanted with PTHL group displayed new bone formation in four weeks without any evidence of residual bacteria, which showed similar antibacterial outcomes to the vancomycin and cancellous bone mixture group. In conclusion, the PTHL composite scaffold is a promising non-antibiotic antimicrobial graft with good biodegradability, biocompatibility, and osteogenic capability for infected bone defects.

Bacterial nanocellulose and long-chain fatty acids interaction

Chronic wounds are a big challenge in contemporary society, as they lead to a decrease in life-quality, amputations and even death. Infections and biofilm formation might occur with chronic wounds, due to the higher susceptibility to antibiotic multi-resistant bacteria. In this situation, novel wound dressing biomaterials are needed for treatment. Thus, the aim of this research was to evaluate a possible BNC interaction with tucumã oil/butter-derived fatty acids, as this system could be a promising biomaterial for wound treating. The interaction between cellobiose (BNC basic unit) and four fatty acids was evaluated by ab initio simulations and density functional theory (DFT), through SIESTA code. Molecular docking was also used to investigate the effect of a possible releasing of the studied fatty acids to the quorum-sensing proteins of Pseudomonas aeruginosa (gram-negative bacterium) and Staphylococcus aureus (gram-positive bacterium). According to ab initio simulations, the interaction between cellobiose and fatty acids derived from tucumã oil/butter was suggested due to physical adsorption (energy around 0.17-1.33 eV) of the lipidic structures into cellobiose. A great binding affinity (∆G ranging from 4.2-8.2 kcal.mol-1) was observed for both protonated and deprotonated fatty acids against P. aeruginosa (LasI, LasA and Rhlr) and S. aureus (ArgA and ArgC) quorum-sensing proteins, indicating that these bioactive compounds might act as potential antimicrobial and/or antibiofilm agents in the proposed system. Hence, from a theoretical viewpoint, the proposed system could be a promising raw biomaterial in the production of chronic wound dressings.

Bacteriophage Genetic Edition Using LSTM.

Bacteriophages are gaining increasing interest as antimicrobial tools, largely due to the emergence of multi-antibiotic-resistant bacteria. Although their huge diversity and virulence make them particularly attractive for targeting a wide range of bacterial pathogens, it is difficult to select suitable phages due to their high specificity which limits their host range. In addition, other challenges remain such as structural fragility under certain environmental conditions, immunogenicity of phage therapy, or development of bacterial resistance. The use of genetically engineered phages may reduce characteristics that hinder prophylactic and therapeutic applications of phages. Nowadays, there is no systematic method to modify a given phage genome conferring its sought characteristics. We explore the use of artificial intelligence for this purpose as it has the potential to both guide and accelerate genome modification to generate phage variants with unique properties that overcome the limitations of natural phages. We propose an original architecture composed of two deep learning-driven components: a phage-bacterium interaction predictor and a phage genome-sequence generator. The former is a multi-branch 1-D convolutional neural network (1D-CNN) that analyses phage and bacterial genomes to predict interactions. The latter is a recurrent neural network, more particularly a long short-term memory (LSTM), that performs genomic modifications to a phage to offer substantial host range improvement. For this component, we developed two different architectures composed of one or two stacked LSTM layers with 256 neurons each. These generators are used to modify, more precisely to rewrite, the genome sequence of 42 selected phages, while the predictor is used to estimate the host range of the modified bacteriophages across 46 strains of Pseudomonas aeruginosa. The proposed generators, trained with an average accuracy of 96.1%, are able to improve the host range for an average of 18 phages among the 42 under study, increasing both their average host range, by 73.0 and 103.7%, and the maximum host ranges from 21 to 24 and 29, respectively. These promising results showed that the use of deep learning methodologies allows genetic modification of phages to extend, for instance, their host range, confirming the potential of these approaches to guide bacteriophage engineering.

An emerging unrated mobile reservoir for antibiotic resistant genes: Does transportation matter to the spread

The regional distribution of antibiotic resistance genes has been caused by the use and preference of antibiotics. Not only environmental factors, but also the population movement associated with transportation development might have had a great impact, but yet less is known regarding this issue. This research study has investigated and reported that the high-speed railway train was a possible mobile reservoir of bacteria with antibiotic resistance, based on the occurrence, diversity, and abundance of antibiotic resistant bacteria (ARB), antibiotic resistance genes (ARGs), and mobile gene elements (MGEs) in untreated train wastewater. High-throughput 16S rRNA sequencing analyses have indicated that opportunistic pathogens like Pseudomonas and Enterococcuss were the predominant bacteria in all samples, especially in cultivable multi-antibiotic resistant bacteria. The further isolated Enterococcus faecalis and Enterococcus faecium exhibited multi-antibiotic resistance ability, potentially being an indicator for disinfection proficiency. Positive correlations amongst ARGs and MGEs were observed, such as between intI1 and tetW, tetA, blaTEM, among Tn916/154 and mefA/F, qnrS, implying a broad dissemination of multi-ARGs during transportation. The study findings suggested that the high-speed railway train wastewater encompassed highly abundant antibiotic-resistant pathogens, and the wastewater discharge without effective treatment may pose severe hazards to human health and ecosystem safety.

Towards efficient immunotherapy for bacterial infection.

The emergence of multiantibiotic-resistant bacteria, often referred to as superbugs, is leading to infections that are increasingly difficult to treat. Further, bacteria have evolved mechanisms by which they subvert the immune response, meaning that even antibiotic-sensitive bacteria can persist through antibiotic therapy. For these reasons, a broad range of viable therapeutic alternatives or conjunctions to traditional antimicrobial therapy are urgently required to reduce the burden of disease threatened by antibiotic resistance. Immunotherapy has emerged as a leading treatment option in cancer, and researchers are now attempting to apply this to infectious disease. This review summarizes and discusses the recent advances in the field and highlights current and future perspectives of using immunotherapies to treat bacterial infections.

Antimicrobial effect of Jatropha tanjorensis leaves on multi antibiotic resistant bacterial isolated from poultry droppings

The widespread use of antibiotics as prophylactics and therapeutic agents has increased the risk of emergence and spread of resistant bacteria leading to prolonged and expensive treatment and greater risk of death in humans. The aim of this study was to determine the antimicrobial effect of the methanol extract of Jatropha tanjorensis leaves on multi antibiotic resistant bacteria isolated from poultry droppings. Multi antibiotic resistant bacterial isolates from poultry dropping already identified from previous research were used. The plant extract was obtained by soxhlet extraction process, susceptibility test was caried out using well in agar diffusion method. The percentage yield of the plant extract using soxhlet extraction process with methanol was 22.84%. The qualitative phytochemical analysis revealed the presence of bioactive compounds such as tannins, flavonoids, cardenolide, steroids, terpenoids, anthraquinone, alkaloids, fixed oil and carbohydrate. The extract showed competitive zones of inhibition on Escherichia fergusonii, Stenotrophomonas maltophilia, Enterobacter clocae, Lysinibacillus sphaericus and Salmonella typhimerium. Enterobacter clocae had the highest zone of inhibition of 17mm at a concentration of 100mg/ml, 10mm at 80mg/ml and 8mm at 50 mg/ml. Stenotrophomonas maltophilia had the lowest zone of inhibition of 10mm at 100mg/ml, 8mm at 80mg/l and 7mm at 50mg/ml. These zones of inhibitions show the antimicrobial potency of Jatropha tajorensis on antibiotic resistant bacteria. Compared with synthetic antibiotics or inorganic chemicals, this plant-derived extract is natural, less toxic, typically residue free with high rate of metabolism. The result of this study gives credence to claims of the medicinal and nutritional properties of Jatropha tanjorensis.

Morphological and molecular identification of multi-antibiotic resistant bacteria in the wound site of diabetic ulcers

Abstract. Risqiyah W, Narulita E, Rofiqoh A, Ludfi AS, Iqbal M. 2021. Morphological and molecular identification of multi-antibiotic resistant bacteria in the wound site of diabetic ulcers. Biodiversitas 23: 663-670. The study aimed to determine the characteristics of diverse species of bacteria found in diabetic ulcers and the inhibition of antibiotics against these bacteria. The method used is Gram staining for morphological characterization, identification using 16S rRNA gene, and pour plate for antibiotic resistance test. The morphological characterization result showed that the colonies had white color, circular shape, flat elevation, entire edge, small size, and basil shape with Gram negative type of bacteria. Another result showed yellow, greenish, pellucid, and green color, irregular shape, raised elevation, undulate and lobate edge, medium size, coccus and Gram positive type of bacteria. Query cover of molecular identification showed 73-100% and 77.61-96.77% for the result of similarity identification with Alcaligenes faecalis, Shigella flexneri, Enterococcus faecalis, Proteus mirabilis, and Acinetobacter seohaensis. The identification and antibiotic resistance testing showed that the bacterial species found in diabetic ulcers was A. faecalis strain NRBC 13111, which was resistant to all tested antibiotics, except A. faecalis strain NRBC 13111 from the UB 2.3K and A. faecalis strain NRBC 13111 from the sample UB 3.4M. Enterococcus faecalis strain ATCC 19433 was resistant to ceftriaxone, ceftazidime, clindamycin and metronidazole, intermediates to cefoperazone, and sensitive to ciprofloxacin. Proteus mirabilis strain JCM 1669 was resistant to clindamycin and metronidazole, sensitive to ceftriaxone and ciprofloxacin, intermediates to ceftazidime and cefoperazone. Proteus mirabilis strain ATCC 29906, A. seohaensis strain SW-100 and S. flexneri strain ATCC 29903 were resistant to all tested antibiotics.

Illustration on Phenotypic and Genotypic Characteristics of Typical Multi-Antibiotic Resistant Bacteria in Aquatic Environments Through Complete Genomes and Comparative Genomics

Illustration on Phenotypic and Genotypic Characteristics of Typical Multi-Antibiotic Resistant Bacteria in Aquatic Environments Through Complete Genomes and Comparative Genomics

Optimizing CSP1 analogs for modulating quorum sensing in Streptococcus pneumoniae with bulky, hydrophobic nonproteogenic amino acid substitutions.

The prompt appearance of multiantibiotic-resistant bacteria necessitates finding alternative treatments that can attenuate bacterial infections while minimizing the rate of antibiotic resistance development. Streptococcus pneumoniae, a notorious human pathogen, is responsible for severe antibiotic-resistant infections. Its pathogenicity is influenced by a cell-density communication system, termed quorum sensing (QS). As a result, controlling QS through the development of peptide-based QS modulators may serve to attenuate pneumococcal infections. Herein, we set out to evaluate the impact of the introduction of bulkier, nonproteogenic side-chain residues on the hydrophobic binding face of CSP1 to optimize receptor-binding interactions in both of the S. pneumoniae specificity groups. Our results indicate that these substitutions optimize the peptide–protein binding interactions, yielding several pneumococcal QS modulators with high potency. Moreover, pharmacological evaluation of lead analogs revealed that the incorporation of nonproteogenic amino acids increased the peptides’ half-life towards enzymatic degradation while remaining nontoxic. Overall, our data convey key considerations for SAR using nonproteogenic amino acids, which provide analogs with better pharmacological properties.

Identification of multi-antibiotic resistant bacteria isolated from Vietnamese climbing perch (Anabas testudineus) on fish farms in Ho Chi Minh City, Vietnam

Climbing perch (Anabas testudineus) is considered a cultured species with high economic value and is widely distributed in several Asian countries, including Vietnam. In climbing perch aquaculture, antibiotic and antibacterial agents have been overused, leading to the emergence of antibiotic-resistant bacteria or genes in the aquatic environment as well as in the seafood products. These antibiotic-resistant bacteria and/or genes can then be spread to human through the food chain, generating detrimental consequences for public health and environment. In this study, 14 bacterial strains were isolated from Vietnamese climbing perch collected from various fish farms in Ho Chi Minh City which possess abnormal symptoms on the outside and in their organs. These bacterial strains were then subjected to several biochemical tests to characterize their colony morphologies. Besides, gene sequencing was performed and as compared to the 16S rRNA, 4 bacterial strains were identified with a high percentage in similarity, including Pseudomonas aeruginosa (99.7%), Aeromonas caviae (99.93%), Kosakonia sacchari (100%), and Edwardsiella ictaluri (99.44%). Additionally, these bacteria showed high levels of antibiotic resistance against beta-lactam, tetracycline, and others. Taken together, these results showed that the four identified bacterial strains isolated from Vietnamese climbing perch circulated in fish farms in Ho Chi Minh City. Their current status of multi-antibiotic resistance indicates further alternative sources of antimicrobial agents, for example, natural compounds, to overcome antibiotic resistance. It is then crucial to identify fish diseases, the associated pathogens, and the new alternatives for antibiotics to protect aquatic ecosystems and water resources.

Multi-antibiotic resistant bacteria in landfill bioaerosols: Environmental conditions and biological risk assessment

Landfills, as well as other waste management facilities are well-known bioaerosols sources. These places may foment antibiotic-resistance in bacterial bioaerosol (A.R.B.) due to inadequate pharmaceutical waste disposal. This issue may foster the necessity of using last-generation antibiotics with extra costs in the health care system, and deaths. The aim of this study was to reveal the multi-antibiotic resistant bacterial bioaerosol emitted by a sanitary landfill and the surrounding area. We evaluated the influence of environmental conditions in the occurrence of A.R.B. and biological risk assessment. Antibiotic resistance found in the bacteria aerosols was compared with the AWaRE consumption classification. We used the BIOGAVAL method to assess the workers' occupational exposure to antibiotic-resistant bacterial bioaerosols in the landfill. This study confirmed the multi-antibiotic resistant in bacterial bioaerosol in a landfill and in the surrounding area. Obtained mean concentrations of bacterial bioaerosols, as well as antibiotic-resistant in bacterial bioaerosol (A.R.B.), were high, especially for fine particles that may be a threat for human health. Results suggest the possible risk of antibiotic-resistance interchange between pathogenic and non-pathogenic species in the landfill facilities, thus promoting antibiotic multi-resistance genes spreading into the environment.

Establishing an invertebrate Galleria mellonella greater wax moth larval model of Neisseria gonorrhoeae infection

ABSTRACT Neisseria gonorrhoeae (gonococcus) causes the human sexually transmitted disease gonorrhea. Studying gonococcal pathogenesis and developing new vaccines and therapies to combat the increasing prevalence of multi-antibiotic resistant bacteria has made use of many ex vivo models based on human cells and tissues, and in vivo vertebrate models, for example, rodent, pig and human. The focus of the current study was to examine the utility of the invertebrate greater wax moth Galleria mellonella as an in vivo model of gonococcal infection. We observed that a threshold of ~106 – 107 gonococci/larva was required to kill >50% of larvae (P < 0.05), and increased toxicity correlated with reduced health index scores and pronounced histopathological changes such as increases in the total lesion grade, melanized nodules, hemocyte reaction, and multifocal adipose body degeneration. Larval death was independent of the expression of pilus or Opa protein or LOS sialylation within a single gonococcal species studied, but the model could demonstrate relative toxicity of different isolates. N. meningitidis, N. lacatamica and gonococci all killed larvae equally, but were significantly less toxic (P > 0.05) than Pseudomonas aeruginosa. Larvae primed with nontoxic doses of gonococci were more susceptible to subsequent challenge with homologous and heterologous bacteria, and larval survival was significantly reduced (P < 0.05) in infected larvae after depletion of their hemocytes with clodronate-liposomes. The model was used to test the anti-gonococcal properties of antibiotics and novel antimicrobials. Ceftriaxone (P < 0.05) protected larvae from infection with different gonococcal isolates, but not azithromycin or monocaprin or ligand-coated silver nanoclusters (P > 0.05).

Combining Functional Genomics and Whole-Genome Sequencing to Detect Antibiotic Resistance Genes in Bacterial Strains Co-Occurring Simultaneously in a Brazilian Hospital.

(1) Background: The rise of multi-antibiotic resistant bacteria represents an emergent threat to human health. Here, we investigate antibiotic resistance mechanisms in bacteria of several species isolated from an intensive care unit in Brazil. (2) Methods: We used whole-genome analysis to identify antibiotic resistance genes (ARGs) and plasmids in 34 strains of Gram-negative and Gram-positive bacteria, providing the first genomic description of Morganella morganii and Ralstonia mannitolilytica clinical isolates from South America. (3) Results: We identified a high abundance of beta-lactamase genes in resistant organisms, including seven extended-spectrum beta-lactamases (OXA-1, OXA-10, CTX-M-1, KPC, TEM, HYDRO, BLP) shared between organisms from different species. Additionally, we identified several ARG-carrying plasmids indicating the potential for a fast transmission of resistance mechanism between bacterial strains. Furthermore, we uncovered two pairs of (near) identical plasmids exhibiting multi-drug resistance. Finally, since many highly resistant strains carry several different ARGs, we used functional genomics to investigate which of them were indeed functional. In this sense, for three bacterial strains (Escherichia coli, Klebsiella pneumoniae, and M. morganii), we identified six beta-lactamase genes out of 15 predicted in silico as those mainly responsible for the resistance mechanisms observed, corroborating the existence of redundant resistance mechanisms in these organisms. (4) Conclusions: Systematic studies similar to the one presented here should help to prevent outbreaks of novel multidrug-resistant bacteria in healthcare facilities.