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Keywords = icaC virulence genes

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20 pages, 3773 KB  
Article
Integrated In Vitro and In Silico Evaluation of Benzimidazolium Salts: Antioxidant Activity, Anti-MRSA/MSSA Effects, and Antibacterial Gene Expression Analysis
by Başak Bedir, Hakan Ünver, Mehmet Çimentepe, Özge Öztürk Çimentepe, Akın Yiğin and Metin Yildirim
Antibiotics 2026, 15(6), 567; https://doi.org/10.3390/antibiotics15060567 - 2 Jun 2026
Viewed by 491
Abstract
Background/Objectives: The emergence of methicillin-resistant Staphylococcus aureus (MRSA) has created an urgent need for the development of novel antimicrobial agents. This study aimed to synthesize and characterize a series of benzimidazolium salts and evaluate their antibacterial, antibiofilm, and molecular interaction properties against MRSA [...] Read more.
Background/Objectives: The emergence of methicillin-resistant Staphylococcus aureus (MRSA) has created an urgent need for the development of novel antimicrobial agents. This study aimed to synthesize and characterize a series of benzimidazolium salts and evaluate their antibacterial, antibiofilm, and molecular interaction properties against MRSA and methicillin-susceptible Staphylococcus aureus (MSSA). Methods: Five benzimidazolium salts bearing different substituents were synthesized and characterized by NMR and FTIR analyses. Their antibacterial activities against MRSA and MSSA were investigated using the resazurin-based minimum inhibitory concentration (MIC) assay and agar diffusion method. Antibiofilm activity was evaluated using crystal violet staining, while scanning electron microscopy (SEM) was employed to examine bacterial morphological changes. Gene expression analysis was performed to assess the effects of the most active compound on virulence- and resistance-related genes. In addition, molecular docking studies were conducted against four target proteins (1MWT, 3ZG5, 1JIJ, and 2Y2M). Results: Among the synthesized compounds, compound 1c exhibited the strongest antibacterial activity, with MIC values of 0.25 µg/mL against MRSA and 0.5 µg/mL against MSSA. It also produced the largest inhibition zone diameter (31.0 ± 0.5 mm). SEM analysis revealed significant morphological alterations in treated bacterial cells, indicating disruption of cellular integrity. Gene expression studies demonstrated that compound 1c downregulated several virulence- and resistance-associated genes, including icaA, dltA, dltB, sarA, norA, and norB. All compounds displayed antibiofilm activity, with compound 1c showing the highest inhibition rate (68.6 ± 0.6%). Molecular docking analysis revealed that compound 1c exhibited the strongest binding affinity toward the 2Y2M target protein, with a docking score of −5.322 kcal/mol. Conclusions: The findings demonstrate that benzimidazolium derivatives, particularly compound 1c, possess potent antibacterial and antibiofilm activities against S. aureus strains and effectively modulate virulence- and resistance-related gene expression. Combined with favorable molecular docking results, these compounds represent promising candidates for the development of new therapeutic agents against resistant S. aureus infections. Full article
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15 pages, 1592 KB  
Article
Antibiotic Resistance and Molecular Characterization of Staphylococcus aureus Strains Colonizing the Nose and Pharynx
by Samuel González-García, Anaíd Bustos-Hamdan, Aída Hamdan-Partida and Jaime Bustos-Martínez
Microorganisms 2025, 13(9), 1978; https://doi.org/10.3390/microorganisms13091978 - 25 Aug 2025
Cited by 2 | Viewed by 2199
Abstract
Staphylococcus aureus colonizes the nasal and pharyngeal mucosa of a considerable proportion of the human population, representing a relevant reservoir for the dissemination of antibiotic resistance. This study aimed to determine the prevalence, antibiotic resistance profiles, and molecular characterization of S. aureus strains [...] Read more.
Staphylococcus aureus colonizes the nasal and pharyngeal mucosa of a considerable proportion of the human population, representing a relevant reservoir for the dissemination of antibiotic resistance. This study aimed to determine the prevalence, antibiotic resistance profiles, and molecular characterization of S. aureus strains isolated from the nose and pharynx of healthy individuals of Mexico City, Mexico. Nasal and pharyngeal swabs were taken from 1777 individuals aged between 1 and 99 years. Microbiological identification, antibiotic susceptibility testing, virulence gene detection, SCCmec typing, and spa-typing allowed the characterization of 1459 strains. S. aureus colonization was found in 59.7% of the population, with more of these strains being found in the pharynx than in the nose. MRSA constituted 9.25% of the strains, with a predominance of strains with SCCmec types IV and IVa. The most frequent resistance of S. aureus strains was to penicillin (87.6%), clindamycin (43.4%) and erythromycin (27.2%), with no statistical differences regarding the anatomical sites of isolation. The most frequently found virulence genes were lukE-D, icaA, sdrC, coa, and gyrA. Sixty-two spa-types were identified, and t-189 was the most common. These findings reflect a high colonization rate and genetic diversity, highlighting the importance of considering both anatomical sites in S. aureus surveillance studies. Full article
(This article belongs to the Special Issue Drug Resistance and Molecular Research of Staphylococcus spp.)
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17 pages, 12673 KB  
Article
Identification of Novel Staphylococcus aureus Core and Accessory Virulence Patterns in Chronic Rhinosinusitis
by Simon P. Goldie, Laurie C. Lau, Huw A. S. Jones, Philip G. Harries, Andrew F. Walls and Rami J. Salib
Int. J. Mol. Sci. 2025, 26(8), 3711; https://doi.org/10.3390/ijms26083711 - 14 Apr 2025
Cited by 5 | Viewed by 2062
Abstract
Staphylococcus aureus (S. aureus) colonizes the nasal cavities of both healthy individuals and patients with chronic rhinosinusitis (CRS) with (CRSwNP) and without (CRSsNP) nasal polyps. Treatment-resistant S. aureus biofilms and intracellular persistence are common in CRS patients, requiring the expression of [...] Read more.
Staphylococcus aureus (S. aureus) colonizes the nasal cavities of both healthy individuals and patients with chronic rhinosinusitis (CRS) with (CRSwNP) and without (CRSsNP) nasal polyps. Treatment-resistant S. aureus biofilms and intracellular persistence are common in CRS patients, requiring the expression of specific virulence factor genes to transition into these forms. We hypothesized that S. aureus isolates from non-diseased controls, CRSsNP patients, and CRSwNP patients would exhibit distinct virulence factor patterns contributing to persistence and intracellular survival in CRS patients. Nasal swabs from seventy-seven individuals yielded S. aureus cultures in eight non-diseased controls, eight CRSsNP patients, and five CRSwNP patients. Whole-genome sequencing analyzed stress, antimicrobial resistance, and virulence genes, including plasmids and prophages. Four virulence factor gene patterns emerged: a core set (hlgA, icaC, hlgB, hlgC, hld, and aur) present in all isolates, and accessory sets, including the enterotoxin gene cluster (seo, sem, seu, sei, and sen) and a partial/complete invasive virulence factor set (splE, splA, splB, lukE, and lukD) (p = 0.001). CRSwNP isolates exhibited incomplete carriage of the core set, with frequent loss of scn, icaC, and hlgA (p < 0.05). These findings suggest that S. aureus has clusters of virulence factors that may act in concert to support the survival and persistence of the bacteria, resulting in enhanced pathogenicity. This may manifest clinically with resistant disease and refractoriness to antibiotics. Full article
(This article belongs to the Special Issue Mechanisms in Biofilm Formation, Tolerance and Control: 2nd Edition)
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17 pages, 4989 KB  
Article
Searching for Virulence Factors among Staphylococcus lugdunensis Isolates from Orthopedic Infections: Correlation of β-hemolysin, hemolysin III, and slush Genes with Hemolytic Activity and Synergistic Hemolytic Activity
by Stefano Ravaioli, Davide Campoccia, Rasoul Mirzaei, Valentina Mariani, Giulia Bottau, Andrea De Donno, Lucio Montanaro, Pietro Speziale and Carla Renata Arciola
Int. J. Mol. Sci. 2023, 24(21), 15724; https://doi.org/10.3390/ijms242115724 - 29 Oct 2023
Cited by 7 | Viewed by 3069
Abstract
Staphylococcus lugdunensis is an emerging high-virulent pathogen. Here, the presence and expression of virulence genes (icaA, fbl, vwbl, fbpA, slush A, B and C, and genes of the putative β-hemolysin and hemolysin III) and the [...] Read more.
Staphylococcus lugdunensis is an emerging high-virulent pathogen. Here, the presence and expression of virulence genes (icaA, fbl, vwbl, fbpA, slush A, B and C, and genes of the putative β-hemolysin and hemolysin III) and the ability to induce synergistic hemolytic activity and hemolysis after 24, 48 and 72 h were investigated in a collection of twenty-two S. lugdunensis clinical isolates. The collection of isolates, mainly from implant orthopedic infections, had previously been grouped by ribotyping/dendrogram analysis and studied for biofilm matrices, biomasses and antibiotic resistances. Two isolates, constituting a unique small ribogroup sharing the same cluster, exhibited an amplicon size of the slush operon (S. lugdunensis synergistic hemolysin) which was shorter than the expected 977 bp. This outcome can predict the genetic lineage of the S. lugdunensis strains. One isolate (cra1342) presented two deletions: one of 90 bp in slush A and the other of 91 bp in slush B. Another isolate (N860314) showed a single 193 bp deletion, which encompassed part of the slush B terminal sequence and most of slush C. The isolate N860314 was devoid of hemolytic activity after 24 h, and the first consideration was that the deleted region deals with the coding of the active enzymatic site of the slush hemolysin. On the other hand, cra1342 and N860314 isolates with different slush deletions and with hemolytic activity after 24 and 48 h, respectively, could have replaced the hemolytic phenotype through other processes. Full article
(This article belongs to the Special Issue Recent Progress in Antimicrobial Peptides)
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20 pages, 769 KB  
Article
The Prevalence of the Virulence Genes of Staphylococcus aureus in Sickle Cell Disease Patients at KSUMC, Riyadh, Saudi Arabia
by Adel A. Abdulmanea, Naiyf S. Alharbi, Ali M. Somily, Jamal M. Khaled and Farjah H. Algahtani
Antibiotics 2023, 12(7), 1221; https://doi.org/10.3390/antibiotics12071221 - 22 Jul 2023
Cited by 5 | Viewed by 3578
Abstract
Staphylococcus aureus in the blood of sickle cell disease (SCD) patients may result in a significant increase in morbidity and mortality. S. aureus strains contain various virulence characteristics, including the ability to create a variety of toxins and develop drug resistance. The current [...] Read more.
Staphylococcus aureus in the blood of sickle cell disease (SCD) patients may result in a significant increase in morbidity and mortality. S. aureus strains contain various virulence characteristics, including the ability to create a variety of toxins and develop drug resistance. The current study sought to assess the prevalence of S. aureus in SCD patients and to identify the pathogen’s virulence characteristics. Between 2017 and 2021, blood samples and data were collected at King Saud University Medical City (KSUMC) in Riyadh, Saudi Arabia. The Vitek system PCR and gene sequencing methods were used for identification, antibiotic resistance patterns, and genetic analysis. During the study period, 47 S. aureus blood isolates (methicillin-resistant S. aureus (MRSA) 41.6% and non-MRSA 58.4%) were isolated from 2406 SCD patients. The prevalence percentages of virulence genes (finbB, sdrC, sdrD, icaA, coa, nuc, hlg, hla, finbA, clfA, efb, pvl, agr, spa, seb, sea, sec, tst, and sed) among all the isolates from the SCD patients compared with non-SCD patients (control group) were as follows: (100% vs. 100%), (100% vs. 100%), (100% vs. 100%), (100% vs. 87.5%), (100% vs. 81.3%), (100% vs. 100%), (100% vs. 100%), (100% vs. 100%), (97.9% vs. 81.3%), (97.9% vs. 100%), (97.9% vs. 87.5%), (54.3% vs. 56.3%), (46.8% vs. 75%), (42.6% vs. 43.8%), (27.7% vs. 0%), (25.5% vs. 12.5%), (12.8% vs. 6.3%), (4.3% vs. 12.5%), and (4.3% vs. 0%). Regarding the resistance genes (plaZ, mecA, ermA, ermC, tetK, tetM, and ermB) of the S. aureus strains isolated from the SCD patients compared with non-SCD patients (control group), the prevalence percentages were as follows: (100% vs. 100%), (100% vs. 56.3%), (0% vs. 31.3%), (31.9% vs. 18.8%), (40.4% vs. 25%), (0% vs. 0%), and (0% vs. 0%). As for the antibiotic (ampicillin, penicillin, amoxicillin, cefazolin, imipenem, oxacillin, erythromycin, tetracycline, azithromycin, ciprofloxacin, moxifloxacin, and levofloxacin) resistance of the S. aureus strains isolated from the SCD patients compared with non-SCD patients (control group), the prevalence percentages were as follows: (100% vs. 100%), (97.9% vs. 100%), (72.3% vs. 25%), (68.1% vs. 37.5%), (68.1% vs. 25%), (66% vs. 25%), (36.2% vs. 18.8%), (23.4% vs. 12.5%), (19.1% vs. 12.5%), (17% vs. 12.5%), (14.9% vs. 25%), and (10.6% vs. 18.7%). This study concluded that several virulence genes were present in the S. aureus strains recovered from the SCD patients at KSUMC, with all the isolates containing the finbB, sdrC, sdrD, icaA, coa, nuc, hlg, and hla genes. Full article
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16 pages, 5461 KB  
Article
Roles of the Crp/Fnr Family Regulator ArcR in the Hemolysis and Biofilm of Staphylococcus aureus
by Tongtong Fu, Zheng Fan, Yujie Li, Zhoufei Li, Hanqing Zhao, Yanling Feng, Guanhua Xue, Jinghua Cui, Chao Yan, Lin Gan, Junxia Feng, Jing Yuan and Fuping You
Microorganisms 2023, 11(7), 1656; https://doi.org/10.3390/microorganisms11071656 - 25 Jun 2023
Cited by 10 | Viewed by 3792
Abstract
Staphylococcus aureus is an opportunistic human pathogen that is often involved in severe infections such as pneumonia and sepsis in which bacterial virulence factors play a key role. Infections caused by S. aureus are often difficult to eradicate, particularly when they are associated [...] Read more.
Staphylococcus aureus is an opportunistic human pathogen that is often involved in severe infections such as pneumonia and sepsis in which bacterial virulence factors play a key role. Infections caused by S. aureus are often difficult to eradicate, particularly when they are associated with biofilm. The physiological roles of the Crp/Fnr family regulator ArcR are elusive in S. aureus. In this study, it was found that the deletion of arcR increased the hemolytic ability and biofilm formation in S. aureus. Differential gene expression analysis by RNA-seq and real-time quantitative reverse transcription PCR showed that genes associated with hemolytic ability (hla and hlb) and biofilm formation (icaA, icaB, icaC and icaD) were significantly upregulated compared with those in the wild-type strain. The results revealed that ArcR regulated the expression of the hla and ica operon by binding to their promoter regions, respectively. This study provided new insights into the functional importance of ArcR in regulating the virulence and biofilm of S. aureus. Full article
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15 pages, 3357 KB  
Article
Effect of Essential Oil from Lippia origanoides on the Transcriptional Expression of Genes Related to Quorum Sensing, Biofilm Formation, and Virulence of Escherichia coli and Staphylococcus aureus
by Andrés Martínez, Elena E. Stashenko, Rodrigo Torres Sáez, German Zafra and Claudia Ortiz
Antibiotics 2023, 12(5), 845; https://doi.org/10.3390/antibiotics12050845 - 3 May 2023
Cited by 35 | Viewed by 5298
Abstract
Microbial infections resistant to conventional antibiotics constitute one of the most important causes of mortality in the world. In some bacterial species, such as Escherichia coli and Staphylococcus aureus pathogens, biofilm formation can favor their antimicrobial resistance. These biofilm-forming bacteria produce a compact [...] Read more.
Microbial infections resistant to conventional antibiotics constitute one of the most important causes of mortality in the world. In some bacterial species, such as Escherichia coli and Staphylococcus aureus pathogens, biofilm formation can favor their antimicrobial resistance. These biofilm-forming bacteria produce a compact and protective matrix, allowing their adherence and colonization to different surfaces, and contributing to resistance, recurrence, and chronicity of the infections. Therefore, different therapeutic alternatives have been investigated to interrupt both cellular communication routes and biofilm formation. Among these, essential oils (EO) from Lippia origanoides thymol-carvacrol II chemotype (LOTC II) plants have demonstrated biological activity against different biofilm-forming pathogenic bacteria. In this work, we determined the effect of LOTC II EO on the expression of genes associated with quorum sensing (QS) communication, biofilm formation, and virulence of E. coli ATCC 25922 and S. aureus ATCC 29213. This EO was found to have high efficacy against biofilm formation, decreasing—by negative regulation—the expression of genes involved in motility (fimH), adherence and cellular aggregation (csgD), and exopolysaccharide production (pgaC) in E. coli. In addition, this effect was also determined in S. aureus where the L. origanoides EO diminished the expression of genes involved in QS communication (agrA), production of exopolysaccharides by PIA/PNG (icaA), synthesis of alpha hemolysin (hla), transcriptional regulators of the production of extracellular toxins (RNA III), QS and biofilm formation transcriptional regulators (sarA) and global regulators of biofilm formation (rbf and aur). Positive regulation was observed on the expression of genes encoding inhibitors of biofilm formation (e.g., sdiA and ariR). These findings suggest that LOTCII EO can affect biological pathways associated with QS communication, biofilm formation, and virulence of E. coli and S. aureus at subinhibitory concentrations and could be a promising candidate as a natural antibacterial alternative to conventional antibiotics. Full article
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12 pages, 590 KB  
Article
Molecular Characterizations of the Coagulase-Negative Staphylococci Species Causing Urinary Tract Infection in Tanzania: A Laboratory-Based Cross-Sectional Study
by Shukrani Phillip, Martha F. Mushi, Arun Gonzales Decano, Jeremiah Seni, Blandina T. Mmbaga, Happiness Kumburu, Eveline T. Konje, Joseph R. Mwanga, Benson R. Kidenya, Betrand Msemwa, Stephen Gillespie, Antonio Maldonado-Barragan, Alison Sandeman, Wilber Sabiti, Mathew T. G. Holden and Stephen E. Mshana
Pathogens 2023, 12(2), 180; https://doi.org/10.3390/pathogens12020180 - 24 Jan 2023
Cited by 16 | Viewed by 7862
Abstract
Background: There is a growing body of evidence on the potential involvement of coagulase-negative Staphylococci (CoNS) in causing urinary tract infections (UTIs). The aim of this study was to delineate virulence potential, antimicrobial resistance genes, and sequence types of CoNS isolated from patients [...] Read more.
Background: There is a growing body of evidence on the potential involvement of coagulase-negative Staphylococci (CoNS) in causing urinary tract infections (UTIs). The aim of this study was to delineate virulence potential, antimicrobial resistance genes, and sequence types of CoNS isolated from patients with UTI symptoms and pyuria in Tanzania. Methods: CoNS from patients with UTI symptoms and more than 125 leucocytes/μL were retrieved, subcultured, and whole-genome sequenced. Results: Out of 65 CoNS isolates, 8 species of CoNS were identified; Staphylococcus haemolyticus, n = 27 (41.5%), and Staphylococcus epidermidis, n = 24 (36.9%), were predominant. The majority of S. haemolyticus were sequence type (ST) 30, with 8 new ST138-145 reported, while the majority of S. epidermidis were typed as ST490 with 7 new ST1184-1190 reported. Sixty isolates (92.3%) had either one or multiple antimicrobial resistance genes. The most frequently detected resistance genes were 53 (21%) dfrG, 32 (12.9%) blaZ, and 26 (10.5%) mecA genes conferring resistance to trimethoprim, penicillin, and methicillin, respectively. Out of 65 isolates, 59 (90.8%) had virulence genes associated with UTI, with a predominance of the icaC 47 (46.5%) and icaA 14 (13.9%) genes. Conclusion:S. haemolyticus and S. epidermidis harboring icaC, dfrG, blaZ, and mecA genes were the predominant CoNS causing UTI in Tanzania. Laboratories should carefully interpret the significant bacteriuria due to CoNS in relation to UTI symptoms and pyuria before labeling them as contaminants. Follow-up studies to document the outcome of the treated patients is needed to add more evidence that CoNS are UTI pathogens. Full article
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18 pages, 2885 KB  
Article
Genomics of Staphylococcus aureus Strains Isolated from Infectious and Non-Infectious Ocular Conditions
by Madeeha Afzal, Ajay Kumar Vijay, Fiona Stapleton and Mark D. P. Willcox
Antibiotics 2022, 11(8), 1011; https://doi.org/10.3390/antibiotics11081011 - 27 Jul 2022
Cited by 17 | Viewed by 5260
Abstract
Staphylococcus aureus is a major cause of ocular infectious (corneal infection or microbial keratitis (MK) and conjunctivitis) and non-infectious corneal infiltrative events (niCIE). Despite the significant morbidity associated with these conditions, there is very little data about specific virulence factors associated with the [...] Read more.
Staphylococcus aureus is a major cause of ocular infectious (corneal infection or microbial keratitis (MK) and conjunctivitis) and non-infectious corneal infiltrative events (niCIE). Despite the significant morbidity associated with these conditions, there is very little data about specific virulence factors associated with the pathogenicity of ocular isolates. A set of 25 S. aureus infectious and niCIEs strains isolated from USA and Australia were selected for whole genome sequencing. Sequence types and clonal complexes of S. aureus strains were identified by using multi-locus sequence type (MLST). The presence or absence of 128 virulence genes was determined by using the virulence finder database (VFDB). Differences between infectious (MK + conjunctivitis) and niCIE isolates from USA and Australia for possession of virulence genes were assessed using the chi-square test. The most common sequence types found among ocular isolates were ST5, ST8 while the clonal complexes were CC30 and CC1. Virulence genes involved in adhesion (ebh, clfA, clfB, cna, sdrD, sdrE), immune evasion (chp, esaD, esaE, esxB, esxC, esxD), and serine protease enzymes (splA, splD, splE, splF) were more commonly observed in infectious strains (MK + conjunctivitis) than niCIE strains (p = 0.004). Toxin genes were present in half of infectious (49%, 25/51) and niCIE (51%, 26/51) strains. USA infectious isolates were significantly more likely to possess splC, yent1, set9, set11, set36, set38, set40, lukF-PV, and lukS-PV (p < 0.05) than Australian infectious isolates. MK USA strains were more likely to possesses yent1, set9, set11 than USA conjunctivitis strains (p = 0.04). Conversely USA conjunctivitis strains were more likely to possess set36 set38, set40, lukF-PV, lukS-PV (p = 0.03) than MK USA strains. The ocular strain set was then compared to 10 fully sequenced non-ocular S. aureus strains to identify differences between ocular and non-ocular isolates. Ocular isolates were significantly more likely to possess cna (p = 0.03), icaR (p = 0.01), sea (p = 0.001), set16 (p = 0.01), and set19 (p = 0.03). In contrast non-ocular isolates were more likely to possess icaD (p = 0.007), lukF-PV, lukS-PV (p = 0.01), selq (p = 0.01), set30 (p = 0.01), set32 (p = 0.02), and set36 (p = 0.02). The clones ST5, ST8, CC30, and CC1 among ocular isolates generally reflect circulating non-ocular pathogenic S. aureus strains. The higher rates of genes in infectious and ocular isolates suggest a potential role of these virulence factors in ocular diseases. Full article
(This article belongs to the Special Issue Ocular Surface Infection and Antimicrobials)
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13 pages, 1938 KB  
Article
Phenotypic and Genotypic Detection of Biofilm-Forming Staphylococcus aureus from Different Food Sources in Bangladesh
by Fatimah Muhammad Ballah, Md. Saiful Islam, Md. Liton Rana, Farhana Binte Ferdous, Rokeya Ahmed, Pritom Kumar Pramanik, Jarna Karmoker, Samina Ievy, Md. Abdus Sobur, Mahbubul Pratik Siddique, Mst. Minara Khatun, Marzia Rahman and Md. Tanvir Rahman
Biology 2022, 11(7), 949; https://doi.org/10.3390/biology11070949 - 22 Jun 2022
Cited by 50 | Viewed by 7367
Abstract
Staphylococcus aureus is a major foodborne pathogen. The ability of S. aureus to produce biofilm is a significant virulence factor, triggering its persistence in hostile environments. In this study, we screened a total of 420 different food samples and human hand swabs to [...] Read more.
Staphylococcus aureus is a major foodborne pathogen. The ability of S. aureus to produce biofilm is a significant virulence factor, triggering its persistence in hostile environments. In this study, we screened a total of 420 different food samples and human hand swabs to detect S. aureus and to determine their biofilm formation ability. Samples analyzed were meat, milk, eggs, fish, fast foods, and hand swabs. S. aureus were detected by culturing, staining, biochemical, and PCR. Biofilm formation ability was determined by Congo Red Agar (CRA) plate and Crystal Violet Microtiter Plate (CVMP) tests. The icaA, icaB, icaC, icaD, and bap genes involved in the synthesis of biofilm-forming intracellular adhesion compounds were detected by PCR. About 23.81% (100/420; 95% CI: 14.17–29.98%) of the samples harbored S. aureus, as revealed by detection of the nuc gene. The CRA plate test revealed 20% of S. aureus isolates as strong biofilm producers and 69% and 11% as intermediate and non-biofilm producers, respectively. By the CVMP staining method, 20%, 77%, and 3% of the isolates were found to be strong, intermediate, and non-biofilm producers. Furthermore, 21% of S. aureus isolates carried at least one biofilm-forming gene, where icaA, icaB, icaC, icaD, and bap genes were detected in 15%, 20%, 7%, 20%, and 10% of the S. aureus isolates, respectively. Bivariate analysis showed highly significant correlations (p < 0.001) between any of the two adhesion genes of S. aureus isolates. To the best of our knowledge, this is the first study in Bangladesh describing the detection of biofilm-forming S. aureus from foods and hand swabs using molecular-based evidence. Our findings suggest that food samples should be deemed a potential reservoir of biofilm-forming S. aureus, which indicates a potential public health significance. Full article
(This article belongs to the Special Issue Microbial Contamination and Food Safety)
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10 pages, 495 KB  
Article
Inducible Clindamycin Resistance and Biofilm Production among Staphylococci Isolated from Tertiary Care Hospitals in Nepal
by Sarita Manandhar, Raju Shrestha, Ratna Shova Tuladhar and Sunil Lekhak
Infect. Dis. Rep. 2021, 13(4), 1043-1052; https://doi.org/10.3390/idr13040095 - 7 Dec 2021
Cited by 10 | Viewed by 6766
Abstract
Resistance to antibiotics, biofilm formation and the presence of virulence factors play important roles in increased mortality associated with infection by staphylococci. The macrolide lincosamide streptogramin B (MLSB) family of antibiotics is commonly used to treat infections by methicillin-resistant isolates. Clinical [...] Read more.
Resistance to antibiotics, biofilm formation and the presence of virulence factors play important roles in increased mortality associated with infection by staphylococci. The macrolide lincosamide streptogramin B (MLSB) family of antibiotics is commonly used to treat infections by methicillin-resistant isolates. Clinical failure of clindamycin therapy has been reported due to multiple mechanisms that confer resistance to MLSB. This study aims to find the incidence of different phenotypes of MLSB resistance and biofilm production among staphylococci. A total of 375 staphylococci were isolated from different clinical samples, received from two tertiary care hospitals in Nepal. Methicillin resistance was detected by cefoxitin disc diffusion method and inducible clindamycin resistance by D test, according to CLSI guidelines. Biofilm formation was detected by the tissue culture plate method and PCR was used to detect ica genes. Of the total staphylococci isolates, 161 (42.9%) were Staphylococcus aureus, with 131 (81.4%) methicillin-resistant strains, and 214 (57.1%) isolates were coagulase-negative staphylococci, with 143 (66.8%) methicillin-resistant strains. The overall prevalence of constitutive MLSB (cMLSB) and inducible MLSB (iMLSB) phenotypes was 77 (20.5%) and 87 (23.2%), respectively. Both iMLSB and cMLSB phenotypes predominated in methicillin-resistant isolates. The tissue culture plate method detected biofilm formation in 174 (46.4%) isolates and ica genes in 86 (22.9%) isolates. Among biofilm producing isolates, cMLSB and iMLSB phenotypes were 35 (20.1%) and 27 (15.5%), respectively. The cMLSB and iMLSB were 11 (12.8%) and 19 (22.1%), respectively, in isolates possessing ica genes. Clindamycin resistance in the form of cMLSB and iMLSB, especially among MRSA, emphasizes the need for routine D tests to be performed in the lab. Full article
(This article belongs to the Section Bacterial Diseases)
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17 pages, 9062 KB  
Article
Carvacrol Essential Oil: A Natural Antibiotic against Zoonotic Multidrug-Resistant Staphylococcus Species Isolated from Diseased Livestock and Humans
by Ahmed H. Abed, Esraa F. Hegazy, Sherif A. Omar, Rehab M. Abd El-Baky, Ahmed A. El-Beih, Ahmed Al-Emam, Ahmed M. S. Menshawy and Eman Khalifa
Antibiotics 2021, 10(11), 1328; https://doi.org/10.3390/antibiotics10111328 - 30 Oct 2021
Cited by 24 | Viewed by 4491
Abstract
Staphylococcus species cause diseases in animals and humans. The prevalence and antimicrobial profiles of Staphylococcus spp. in animals and human samples in the Minya Governorate, Egypt, were determined, and resistance- and virulence-associated genes were observed in multidrug-resistant (MDR) isolates. Moreover, the antibacterial effect [...] Read more.
Staphylococcus species cause diseases in animals and humans. The prevalence and antimicrobial profiles of Staphylococcus spp. in animals and human samples in the Minya Governorate, Egypt, were determined, and resistance- and virulence-associated genes were observed in multidrug-resistant (MDR) isolates. Moreover, the antibacterial effect of carvacrol essential oil (EO) on the MDR isolates was studied. A total of 216 samples were aseptically collected from subclinically mastitic cow’s milk (n = 100), sheep abscesses (n = 25) and humans (n = 91). Out of 216 samples, a total of 154 single Staphylococcus species (71.3%) were isolated. The most frequent bacterial isolates were S. aureus (43%), followed by S. schleiferi (25%), S. intermedius (12%), S. xylosus (12%), S. haemolyticus (4.5%), S. epidermidis (2%) and S. aurecularis (1%). Haemolytic activity and biofilm production were detected in 77 and 47% of isolates, respectively. Antimicrobial susceptibility testing showed a high degree of resistance to the most commonly used antimicrobials in human and veterinary practices. The mecA, vanA, vanC1 and ermC resistance genes were detected in 93, 42, 83 and 13% of isolates, respectively. Moreover, hla, icaA and icaD virulence genes were detected in 50, 75 and 78% of isolates, respectively. Carvacrol effectively inhibited the growth of all tested isolates at concentrations of 0.1, 0.05 and 0.04% while a concentration of 0.03% inhibited 75% of isolates. Interestingly, some phenotypic changes were observed upon treatment with a carvacrol oil concentration of 0.03%. All the treated MDR Staphylococcus isolates changed from multidrug resistant to either susceptible or intermediately susceptible to 2–3 antimicrobials more than parental bacterial isolates. Real-time PCR was applied for the detection of the differential expression of mecA and vanC1 genes before and after treatment with carvacrol which revealed a mild reduction in both genes’ expression after treatment. Staphylococcus spp. Containing MDR genes are more likely to spread between humans and animals. From these results, carvacrol EO is a promising natural alternative to conventional antimicrobials for pathogens impacting human health and agriculture due to its potential antimicrobial effect on MDR pathogens; even in sub-lethal doses, carvacrol EO can affect their phenotypic properties and genes’ expression. Full article
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12 pages, 458 KB  
Article
Molecular Characterization of Antimicrobial Resistance and Virulence Genes of Bacterial Pathogens from Bovine and Caprine Mastitis in Northern Lebanon
by Zahie Abboud, Lucia Galuppo, Marco Tolone, Maria Vitale, Roberto Puleio, Marwan Osman, Guido Ruggero Loria and Monzer Hamze
Microorganisms 2021, 9(6), 1148; https://doi.org/10.3390/microorganisms9061148 - 27 May 2021
Cited by 17 | Viewed by 6442
Abstract
Mastitis is an infectious disease encountered in dairy animals worldwide that is currently a growing concern in Lebanon. This study aimed at investigating the etiology of the main mastitis-causing pathogens in Northern Lebanon, determining their antimicrobial susceptibility profiles, and identifying their antimicrobial resistance [...] Read more.
Mastitis is an infectious disease encountered in dairy animals worldwide that is currently a growing concern in Lebanon. This study aimed at investigating the etiology of the main mastitis-causing pathogens in Northern Lebanon, determining their antimicrobial susceptibility profiles, and identifying their antimicrobial resistance (AMR) genes. A total of 101 quarter milk samples were collected from 77 cows and 11 goats presenting symptoms of mastitis on 45 dairy farms. Bacterial identification was carried out through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibility was tested by disc diffusion and broth microdilution methods. Molecular characterization included polymerase chain reaction (PCR) screening for genes encoding extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC among Enterobacterales isolates, and virulence factors among Staphylococcus isolates. Escherichia coli isolates were subjected to phylogenetic typing by a quadruplex PCR method. The most frequently identified species were Streptococcus uberis (19.2%), Streptococcus agalactiae (15.1%), E. coli (12.3%), and Staphylococcus aureus (10.96%). Gram-positive bacteria were resistant to macrolides and tetracycline, whereas gram-negative bacteria displayed resistance to ampicillin and tetracycline. Two ESBL genes, blaTEM (83.3%) and blaOXA (16.7%), and one AmpC beta-lactamase gene, blaCMY-II (16.7%), were detected among six E. coli isolates, which mainly belonged to phylogenetic group B1. Among Staphylococcus spp., the mecA gene was present in three isolates. Furthermore, four isolates contained at least one toxin gene, and all S. aureus isolates carried the ica operon. These findings revealed the alarming risk of AMR in the Lebanese dairy chain and the importance of monitoring antimicrobial usage. Full article
(This article belongs to the Special Issue ß-Lactamases)
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18 pages, 1312 KB  
Article
Virulence Factors in Staphylococcus Associated with Small Ruminant Mastitis: Biofilm Production and Antimicrobial Resistance Genes
by Nara Cavalcanti Andrade, Marta Laranjo, Mateus Matiuzzi Costa and Maria Cristina Queiroga
Antibiotics 2021, 10(6), 633; https://doi.org/10.3390/antibiotics10060633 - 25 May 2021
Cited by 43 | Viewed by 6510
Abstract
Small ruminant mastitis is a serious problem, mainly caused by Staphylococcus spp. Different virulence factors affect mastitis pathogenesis. The aim of this study was to investigate virulence factors genes for biofilm production and antimicrobial resistance to β-lactams and tetracyclines in 137 staphylococcal isolates [...] Read more.
Small ruminant mastitis is a serious problem, mainly caused by Staphylococcus spp. Different virulence factors affect mastitis pathogenesis. The aim of this study was to investigate virulence factors genes for biofilm production and antimicrobial resistance to β-lactams and tetracyclines in 137 staphylococcal isolates from goats (86) and sheep (51). The presence of coa, nuc, bap, icaA, icaD, blaZ, mecA, mecC, tetK, and tetM genes was investigated. The nuc gene was detected in all S. aureus isolates and in some coagulase-negative staphylococci (CNS). None of the S. aureus isolates carried the bap gene, while 8 out of 18 CNS harbored this gene. The icaA gene was detected in S. aureus and S. warneri, while icaD only in S. aureus. None of the isolates carrying the bap gene harbored the ica genes. None of the biofilm-associated genes were detected in 14 isolates (six S. aureus and eight CNS). An association was found between Staphylococcus species and resistance to some antibiotics and between antimicrobial resistance and animal species. Nine penicillin-susceptible isolates exhibited the blaZ gene, questioning the reliability of susceptibility testing. Most S. aureus isolates were susceptible to tetracycline, and no cefazolin or gentamycin resistance was detected. These should replace other currently used antimicrobials. Full article
(This article belongs to the Special Issue Antimicrobial Resistance and Virulence - 2nd Volume)
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14 pages, 5812 KB  
Article
Virulence Factors Found in Nasal Colonization and Infection of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolates and Their Ability to Form a Biofilm
by Thamiris Santana Machado, Felipe Ramos Pinheiro, Lialyz Soares Pereira Andre, Renata Freire Alves Pereira, Reginaldo Fernandes Correa, Gabriela Coutinho de Mello, Tainara Aparecida Nunes Ribeiro, Bruno Penna, Daniela Sachs and Fábio Aguiar-Alves
Toxins 2021, 13(1), 14; https://doi.org/10.3390/toxins13010014 - 25 Dec 2020
Cited by 17 | Viewed by 4454
Abstract
Hospitalizations related to Methicillin-resistant Staphylococcus aureus (MRSA) are frequent, increasing mortality and health costs. In this way, this study aimed to compare the genotypic and phenotypic characteristics of MRSA isolates that colonize and infect patients seen at two hospitals in the city of [...] Read more.
Hospitalizations related to Methicillin-resistant Staphylococcus aureus (MRSA) are frequent, increasing mortality and health costs. In this way, this study aimed to compare the genotypic and phenotypic characteristics of MRSA isolates that colonize and infect patients seen at two hospitals in the city of Niterói—Rio de Janeiro, Brazil. A total of 147 samples collected between March 2013 and December 2015 were phenotyped and genotyped to identify the protein A (SPA) gene, the mec staphylococcal chromosomal cassette (SCCmec), mecA, Panton-Valentine Leucocidin (PVL), icaC, icaR, ACME, and hla virulence genes. The strength of biofilm formation has also been exploited. The prevalence of SCCmec type IV (77.1%) was observed in the colonization group; however, in the invasive infection group, SCCmec type II was prevalent (62.9%). The Multilocus Sequence Typing (MLST), ST5/ST30, and ST5/ST239 analyses were the most frequent clones in colonization, and invasive infection isolates, respectively. Among the isolates selected to assess the ability to form a biofilm, 51.06% were classified as strong biofilm builders. Surprisingly, we observed that isolates other than the Brazilian Epidemic Clone (BEC) have appeared in Brazilian hospitals. The virulence profile has changed among these isolates since the ACME type I and II genes were also identified in this collection. Full article
(This article belongs to the Special Issue Resistance to Staphylococcus aureus Toxins)
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