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Bacterial pathogens cause pain and death, add significantly to the expense of healthcare globally, and pose a serious concern in many aspects of daily life. Additionally, they raise significant issues in other industries, including pharmaceuticals, clothing, and food packaging. Due to their unique properties, a great deal of attention has been given to biogenic metal nanoparticles, nanocomposites, and their applications against pathogenic bacteria. This study is focused on biogenic silver and copper nanoparticles and their composites (UL/Ag2 O-NPS, Ul/CuO-NPs, and Ul/Ag/Cu-NCMs) produced by the marine green alga Ulva lactuca. The characterization of biogenic nanoparticles UL/Ag2 O-NPS and Ul/CuO-NPs and their composites Ul/Ag/Cu-NCMs has been accomplished by FT-IR, SEM, TEM, EDS, XRD, and the zeta potential. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) experiments were conducted to prove antibacterial activity against both Gram-positive and Gram-negative bacteria and anti-biofilm. The FTIR spectroscopy results indicate the exiting band at 1633 cm-1, which represents N-H stretching in nanocomposites, with a small shift in both copper and silver nanoparticles, which is responsible for the bio-reduction of nanoparticles. The TEM image reveals that the Ul/Ag/Cu-NCMs were hexagonal, and the size distribution ranged from 10 to 35 nm. Meanwhile, Ul/CuO-NPs are rod-shaped, whereas UL/Ag2 O-NPS are spherical. The EDX analysis shows that Cu metal was present in a high weight percentage over Ag in the case of bio-Ag/Cu-NCMs. The X-ray diffraction denotes that Ul/Ag/Cu-NCMs, UL/CuO-NPs, and UL/Ag2 O-NPS were crystalline. The results predicted by the zeta potential demonstrate that Ul/Ag/Cu-NCMs were more stable than Ul/CuO-NPs. The antibacterial activity of UL/Ag2 O-NPS, Ul/Ag/Cu-NCMs, and UL/CuO-NPs was studied against eleven Gram-negative and Gram-positive multidrug-resistant bacterial species. The maximum inhibition zones were obtained with UL/Ag2 O-NPS, followed by Ul/Ag/Cu-NCMs and Ul/CuO-NPs in all the tested bacteria. The maximum anti-biofilm percentage formed by E. coli KY856933 was obtained with UL/Ag2 O-NPS. These findings suggest that the synthesized nanoparticles might be a great alternative for use as an antibacterial agent against different multidrug-resistant bacterial strains.

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Zinc oxide nanoparticles have many advantages for nano-biotechnologists due to their intense biomedical applications. ZnO-NPs are used as antibacterial agents, which influence bacterial cells through the rupture of the cell membrane and the generation of reactive free radicals. Alginate is a polysaccharide of natural origin due to its excellent properties that are used in various biomedical applications. Brown algae are good sources of alginate and are used as a reducing agent in the synthesis of nanoparticles. This study aims to synthesize ZnO-NPs by using brown alga Fucus vesiculosus (Fu/ZnO-NPs) and also to extract alginate from the same alga, which is used in coating the ZnO-NPs (Fu/ZnO-Alg-NCMs). The characterizations of Fu/ZnO-NPs and Fu/ZnO-Alg-NCMs were determined by FTIR, TEM, XRD, and zeta potential. The antibacterial activities were applied against multidrug resistance bacteria of both gram-positive and negative. The results obtained in FT-TR showed there are some shifts in the peak positions of Fu/ZnO-NPs and Fu/ZnO-Alg-NCMs. The peak at 1655 cm-1, which assigned amide I-III, is present in both Fu/ZnO-NPs and Fu-Alg-ZnO-NCMs; this band is responsible for bio-reductions and stabilization of both nanoparticles. The TEM images proved the Fu/ZnO-NPs have rod shapes with sizes ranging from 12.68 to 17.66 and are aggregated, but Fu/ZnO/Alg-NCMs are spherical in shape with sizes ranging from 12.13 to 19.77. XRD-cleared Fu/ZnO-NPs have nine sharp peaks that are considered good crystalline, but Fu/ZnO-Alg-NCMs have four broad and sharp peaks that are considered semi-crystalline. Both Fu/ZnO-NPs and Fu/ZnO-Alg-NCMs have negative charges (-1.74 and -3.56, respectively). Fu/ZnO-NPs have more antibacterial activities than Fu/ZnO/Alg-NCMs in all tested multidrug-resistant bacterial strains. Fu/ZnO/Alg-NCMs had no effect on Acinetobacter KY856930, Staphylococcus epidermidis, and Enterobacter aerogenes, whereas there was an apparent effect of ZnO-NPs against the same strains.

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Pathogens resistant to antimicrobials form a significant threat to public health worldwide. Tackling multidrug-resistant pathogens via screening metagenomic libraries has become a common approach for the discovery of new antibiotics from uncultured microorganisms. This study focuses on capturing nonribosomal peptide synthase (NRPS) gene clusters implicated in the synthesis of many natural compounds of industrial relevance. A NRPS PCR assay was used to screen 2976 Escherichia coli clones in a soil metagenomic library to target NRPS genes. DNA extracts from 4 clones were sequenced and subjected to bioinformatic analysis to identify NRPS domains, their phylogeny, and substrate specificity.Successfully, 17 NRPS-positive hits with a biosynthetic potential were identified. DNA sequencing and BLAST analysis confirmed that NRPS protein sequences shared similarities with members of the genus Delftia in the Proteobacteria taxonomic position. Multiple alignment and phylogenetic analysis demonstrated that clones no. 15cd35 and 15cd37 shared low bootstrap values (54%) and were distantly far from close phylogenetic neighbors. Additionally, NRPS domain substrate specificity has no hits with the known ones; hence, they are more likely to use different substrates to produce new diverse antimicrobials. Further analysis confirmed that the NRPS hits resemble several transposon elements from other bacterial taxa, confirming its diversity. We confirmed that the analyses of the soil metagenomic library revealed a diverse set of NRPS related to the genus Delftia. An in-depth understanding of those positive NRPS hits is a crucial step for genetic manipulation of NRPS, shedding light on alternative novel antimicrobial compounds that can be used in drug discovery and hence supports the pharmaceutical sector.

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One of the most prevalent chronic infectious disorders is tooth decay. Acids produced when plaque bacteria break down sugar in the mouth cause tooth decay. Streptococcus mutans and Lactobacillus acidophilus are the most prominent species related to dental caries. Innovative biocidal agents that integrate with a biomaterial to prevent bacterial colonization have shown remarkable promise as a result of the rapid advancement of nanoscience and nanotechnology. In this study, Ulva lactuca was used as a cellulose source and reducing agent to synthesize nanocellulose and Ulva/Ag/cellulose/nanocomposites. The characterizations of nanocellulose and Ulva/Ag/cellulose/nanocomposites were tested for FT-IR, TEM, SEM, EDS, XRD, and zeta potential. Ulva/Ag/cellulose/nanocomposites and Ulva/nanocellulose, both blended with fluoride, were tested as an antibacterial against S. mutans ATCC 25175 and L. acidophilus CH-2. The results of the SEM proved that nanocellulose is filament-shaped, and FT-IR proved that the functional groups of Ulva/nanocellulose and Ulva/Ag/cellulose/nanocomposites and cellulose are relatively similar but present some small diffusion in peaks. The TEM image demonstrated that the more piratical size distribution of Ulva/Ag/cellulose/nanocomposites ranged from 15 to 20 nm, and Ulva/nanocellulose ranged from 10 to 15 nm. Ulva/Ag/cellulose/nanocomposites have higher negativity than Ulva/nanocellulose. Ulva/Ag/cellulose/nanocomposites and Ulva/nanocellulose possess antibacterial activity against S. mutans ATCC 25175 and L. acidophilus CH-2, but Ulva/Ag/cellulose/nanocomposites are more effective, followed by that blended with fluoride. It is possible to use Ulva/Ag/cellulose/nanocomposites as an antimicrobial agent when added to toothpaste. It is promising to discover an economic and safe nanocomposite product from a natural source with an antimicrobial agent that might be used against tooth bacteria.

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Dental caries is an infectious oral disease caused by the presence of different bacteria in biofilms. Multidrug resistance (MDR) is a major challenge of dental caries treatment. Swabs were taken from 65 patients with dental caries in Makkah, Saudi Arabia. Swabs were cultivated on mitis salivarius agar and de Man, Rogosa, and Sharpe (MRS) agar. VITEK 2 was used for the identification of isolated bacteria. Antibiotic susceptibility testing of the isolated bacteria was performed using commercial antibiotic disks. Ulva lactuca was used as a reducing agent and cellulose source to create nanocellulose and Ag/cellulose nanocomposites. Fourier-transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray diffraction spectroscopy (XRD) were used to characterize nanocellulose and Ag/cellulose nanocomposites. The results showed that most bacterial isolates were Streptococcus spp., followed by Staphylococcus spp. on mitis salivarius media. Lactobacillus spp. and Corynebacterium group f-1 were the bacterial isolates on de Man, Rogosa, and Sharpe (MRS) media. The antibiotic susceptibility test revealed resistance rates of 77%, 93%, 0, 83%, 79%, and 79% against penicillin G, Augmentin, metronidazole, ampicillin, ciprofloxacin, and cotrimoxazole, respectively. Ag/cellulose nanocomposites and Ag/cellulose nanocomposites with fluoride were the most effective antibacterial agents. The aim of this work was to assess the antibacterial activity of Ag/cellulose nanocomposites with and without fluoride against bacteria isolated from the oral cavities of patients with dental caries. This study demonstrated that Ag/cellulose nanocomposites have antibacterial properties against multidrug-resistant bacteria that cause dental caries.

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The in vitro callus induction of Solanum incanum L. was executed on MS medium supplemented with different concentrations of auxin and cytokinin utilizing petioles and explants of leaves. The highest significant fresh weights from petioles and leaf explants were 4.68 and 5.13 g/jar for the medium supplemented with1.0 mg L-1 BA and 1.0 mg L-1 2,4-D. The callus extract of the leaves was used for the green synthesis of silver nanoparticles (Ag-NPs). Analytical methods used for Ag-NPs characterization were UV-vis spectroscopy, Fourier Transform Infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and Transmission Electron Microscopy (TEM). Spherical, crystallographic Ag-NPs with sizes ranging from 15 to 60nm were successfully formed. The FT-IR spectra exhibited the role of the metabolites involved in callus extract in reducing and capping Ag-NPs. The biological activities of Ag-NPs were dose-dependent. The MIC value for Staphylococcus aureus, Bacillus subtilis, and Escherichia coli was 12.5 µg mL-1, while it was 6.25 µg mL-1 for Klebsiella pneumoniae, Pseudomonas aeruginosa, and Candida albicans. The highest inhibition of phytopathogenic fungi Alternaria alternata, Fusarium oxysporum, Aspergillus niger, and Pythium ultimum was 76.3 ± 3.7, 88.9 ± 4.1, 67.8 ± 2.1, and 76.4 ± 1.0%, respectively at 200 µg mL-1. Moreover, green synthesized Ag-NPs showed cytotoxic efficacy against cancerous cell lines HepG2, MCF-7 and normal Vero cell line with IC50 values of 21.76 ± 0.56, 50.19 ± 1.71, and 129.9 ± 0.94 µg mL-1, respectively.

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BACKGROUND AND AIM: Recently, the extensive use of quinolones led to increased resistance to these antimicrobial agents, with different rates according to the organism and the geographical region. The aim of this study was to detect the resistance rate of Klebsiella pneumoniae Iraqi isolates toward quinolone antimicrobial agents, to determine genetic mutations in gyrA and parC, to screen for efflux-pump activity, and to screen the presence of plasmid-mediated quinolone resistance (PMQR) genes. METHODS: Forty-three K. pneumoniae isolates were confirmed phenotypically and genotypically by Vitek 2 system and species specific primers by PCR using the targeting rpo gene followed by sequencing. Antibiotic susceptibility test was carried out using disc diffusion method. Quinolone resistant isolates were subjected to ciprofloxacin MIC testing, and cartwheel method to screen for efflux pump activity. The presence of the plasmid mediated quinolone resistance genes qepA, qnrB, qnrS, and aac(6)Ib was tested by PCR. Sequencing of gyrA and parC was performed. RESULTS: We observed a high rate of resistance to ceftriaxone, gentamicin ciprofloxacin, and levofloxacin. Low rate of resistance was detected against amikacin and azithromycin. Ciprofloxacin MIC results revealed that 96.1% of the isolates had MICs >256 µg/mL, 83.4% had MICs >512 µg/mL while 34.6% had MIC >1024 µg/mL. Testing of isolates against ciprofloxacin mixed with EtBr at various concentrations resulted in decreased resistant. Sequencing results showed that Ser83Leu was the most common mutation in gyrA that was observed in all quinolone resistant isolates, followed by Asp87Asn. Ser80Ile mutation in parC was observed in 77.7% of the tested isolates. The prevalence of PMQR genes was 92.5% aac (6)-Ib, 51.8% qnrB, 40.7% qepA, and 37% qnrS. CONCLUSION: Quinolone resistance is common in K. pneumoniae isolates in Baghdad. The frequent mutation in gyrA and parC, and the presence of PMQR genes is alarming.

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The population structure of Pseudomonas aeruginosa is panmictic-epidemic in nature, with the prevalence of some high-risk clones. These clones are often linked to virulence, antibiotic resistance, and more morbidity. The clonal success of these lineages has been linked to acquisition and spread of mobile genetic elements. The main aim of the study was to explore other molecular markers that explain their global success. A comprehensive set of 528 completely sequenced P. aeruginosa genomes was analyzed. The population structure was examined using Multilocus Sequence Typing (MLST). Strain relationships analysis and diversity analysis were performed using the geoBURST Full Minimum Spanning Tree (MST) algorithm and hierarchical clustering. A phylogenetic tree was constructed using the Unweighted Pair Group Method with Arithmetic mean (UPGMA) algorithm. A panel of previously investigated resistance markers were examined for their link to high-risk clones. A novel panel of molecular markers has been identified in relation to risky clones including armR, ampR, nalC, nalD, mexZ, mexS, gyrAT83I, gyrAD87N, nalCE153Q, nalCS46A, parCS87W, parCS87L, ampRG283E, ampRM288R, pmrALeu71Arg, pmrBGly423Cys, nuoGA890T, pstBE89Q, phoQY85F, arnAA170T, arnDG206C, and gidBE186A. In addition to mobile genetic elements, chromosomal variants in membrane proteins and efflux pump regulators can play an important role in the success of high-risk clones. Finding risk-associated markers during molecular surveillance necessitates applying more infection-control precautions.

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BACKGROUND AND AIM: Bacillus species are widely distributed microorganisms in nature that are responsible for outbreaks of food poisoning and a common cause of food spoilage. This study aimed to isolate and identify foodborne Bacillus species from meat and to determine the antimicrobial activities of commercial essential oils and spices powder extracted from certain medicinal plants. METHODS: Sixty meat samples were collected in Assiut city and subdivided into raw meat and processed meat. Bacillus spp were isolated and identified according to their cultural characters, biochemical reactions, serological typing, and 16S rRNA gene sequencing. The antibacterial activity of essential oils and spices powder was measured by using well-diffusion and microbial count techniques. RESULTS: The prevalence of Bacillus spp. in the examined raw meat samples and processed meat samples was 13.34%, and 26.67%, respectively. There was a marked decrease in the total Bacillus species count after treatment of minced beef with essential oils and spices powder compared to the untreated one. Black seed oil was the most potent antibacterial essential oil among the tested oils present in this study. CONCLUSION: Essential oils and spices powder of certain medicinal plants (cumin: Cuminum cyminum, black seeds: Nigella sativa, cloves: Syzygium aromaicum, cinnamon: Cinnamomum zeylanicum, and Marjoram: Origanum majorana) have a potential in vitro antimicrobial activity against Bacillus spp. Furthermore, Nigella sativa oil exhibited the most potent antibacterial activity against Bacillus spp.

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PURPOSE: Carbapenems are considered the most efficient antibiotic used in the treatment of nosocomial infections. Carbapenem-resistant Gram-negative rods are becoming a serious hazard in hospitals threatening public health. The aim of the current study was to investigate the prevalence of carbapenem-resistant Gram-negative pathogens incriminated in healthcare-associated infections, along with antimicrobial resistance profiles, carbapenemase and metallo-ß-lactamase production, and their molecular characterization. METHODS: A total of 186 clinical specimens were collected from 133 patients at various hospitals in Cairo, Egypt. The obtained specimens were subjected to bacteriological examination, antimicrobial susceptibility testing, detection of carbapenemase production using the modified Hodge test (MHT), the metallo-ß-lactamase production using the EDTA combined disc test (CDT), and PCR-based detection of the bla KPC and bla GES resistance genes. The identification of the highly resistant retrieved isolates was then confirmed by 16S rRNA gene sequencing. RESULTS: The most common isolated Gram-negative species was Klebsiella pneumoniae (40.9%), followed by Acinetobacter baumannii (18.8%), Pseudomonas aeruginosa (17.3%), Escherichia coli (15.4%), Enterobacter aerogenes (5.3%), and Proteus mirabilis (2.4%). The prevalence of carbapenem-resistant isolates was 36.1% (n=75). However, 86.5% of the recovered clinical isolates were susceptible to colistin. The MHT revealed that 33.6% (n=70) of the tested strains were positive for carbapenemase production, while the CDT showed that 33.17% (n=69) of the examined strains were metallo-ß-lactamase producers. The PCR revealed that 98.6% (74/75) of the tested strains possessed the bla KPC gene; moreover, 97.3% (73/75) of the examined strains harbored the bla GES gene. CONCLUSION: This study displayed the emergence of carbapenem-resistant Gram-negative pathogens incriminated in healthcare-associated infections. The accurate identification of carbapenem-resistant bacterial pathogens is pivotal for the treatment of patients, in addition to propelling appropriate contamination control measures to restrain the fast spread of such pathogens. Colistin showed a potent in vitro antimicrobial activity against the carbapenem-resistant strains.