The aim of this study is to evaluate opportunistic pathogenic bacteria of the genus Pseudomonas in anthropogenically impacted bathing waters, primarily focusing on bathing ponds. The findings include the detection of these bacteria, their susceptibility to selected antibiotics, and the determination of the Exotoxin A (exoA) gene using PCR method. P. aeruginosa was present in most samples, albeit in low concentrations (1-14 CFU/100 mL). The presence of P. otitidis, which is associated with ear infection, in this type of bathing water, was not rare (up to 90 CFU/100 mL). This species would not be detected by the standard methods, including tests on acetamid medium, used for P. aeruginosa in water. The isolated strains of P. otitidis lack the exoA gene and exhibited higher resistance to meropenem compared to P. aeruginosa.
- MeSH
- ADP Ribose Transferases genetics MeSH
- Anti-Bacterial Agents * pharmacology MeSH
- Drug Resistance, Bacterial MeSH
- Bacterial Proteins genetics MeSH
- Bacterial Toxins genetics MeSH
- Pseudomonas aeruginosa Exotoxin A MeSH
- Exotoxins genetics MeSH
- Virulence Factors genetics MeSH
- Microbial Sensitivity Tests * MeSH
- Water Microbiology * MeSH
- Polymerase Chain Reaction MeSH
- Pseudomonas * genetics isolation & purification classification drug effects MeSH
- Ponds * microbiology MeSH
- Publication type
- Journal Article MeSH
Despite the lower virulence of current SARS-CoV-2 variants and high rates of vaccinated and previously infected subjects, COVID-19 remains a persistent threat in kidney transplant recipients (KTRs). This study evaluated the parameters of anti-SARS-CoV-2 antibody production in 120 KTRs. The production of neutralizing antibodies in KTRs, following booster vaccination with the mRNA vaccine BNT162b2, was significantly decreased and their decline was faster than in healthy subjects. Factors predisposing to the downregulation of anti-SARS-CoV-2 neutralizing antibodies included age, lower estimated glomerular filtration rate, and a full dose of mycophenolate mofetil. Neutralizing antibodies correlated with those targeting the SARS-CoV-2 receptor binding domain (RBD), SARS-CoV-2 Spike trimmer, total SARS-CoV-2 S1 protein, as well as with antibodies to the deadly SARS-CoV-1 virus. No cross-reactivity was found with antibodies against seasonal coronaviruses. KTRs exhibited lower postvaccination production of neutralizing antibodies against SARS-CoV-2; however, the specificity of their humoral response did not differ compared to healthy subjects.
- MeSH
- COVID-19 * immunology prevention & control MeSH
- Adult MeSH
- Spike Glycoprotein, Coronavirus immunology MeSH
- Immunity, Humoral MeSH
- Middle Aged MeSH
- Humans MeSH
- Antibodies, Neutralizing * blood immunology MeSH
- Transplant Recipients * MeSH
- Antibodies, Viral * blood immunology MeSH
- SARS-CoV-2 * immunology MeSH
- Immunization, Secondary MeSH
- Aged MeSH
- Kidney Transplantation * adverse effects MeSH
- BNT162 Vaccine immunology administration & dosage MeSH
- COVID-19 Vaccines immunology administration & dosage MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Medical students are exposed to the hospital environment and patients during their studies, increasing the risk of exposure to virulent and antibiotic-resistant isolates of Staphylococcus aureus. The aim of the study is to determine the prevalence of Staphylococcus aureus among medical students who have varying levels of exposure to the hospital environment to provide valuable insights into the risk of colonization and transmission. Nasal swabs and fingerprints were obtained and cultured on a selective medium for staphylococci. The obtained isolates were confirmed as methicillin-sensitive S. aureus (MSSA) or methicillin-resistant (MRSA) using PCR. Antibiotic resistance, the presence of virulence genes including enterotoxin encoding genes, and spa typing were performed. Among pre-clinical students, MSSA was detected on the nose in 45.2% and on the fingerprints in 10.6% of the participants. Among clinical students, MSSA was detected on the nose in 42.0% and on the fingerprints in 25.4%. Only one MRSA isolate was obtained. Genes seg and sei were the most frequently detected in both student groups, with their presence in over 40% of isolates among clinical students. The eta and etb genes were mainly detected from the nose in both student groups. In pre-clinical students, S. aureus carrying eta gene occurred in 6.4% and etb in 8.5%. In clinical students, the occurrence was 5.1% for eta and 8.5% for etb. The tst gene was identified only in the nose and fingerprints of the clinical student group. The most frequently observed resistance was to clindamycin and erythromycin. In total 58 different spa types were identified. High rates of asymptomatic MSSA carriage were observed in both groups of medical students. Detected MSSA strains showed a high degree of genetic variability, with a number of them carrying the virulence and antibiotic resistance genes. Although students do not exhibit increased risk to their patient's, increased hygiene is required in asymptomatic carriage personnel. The overall prevalence of MRSA was low, with a minimal risk of spread.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Adult MeSH
- Virulence Factors * genetics MeSH
- Humans MeSH
- Methicillin-Resistant Staphylococcus aureus genetics isolation & purification drug effects classification MeSH
- Microbial Sensitivity Tests MeSH
- Young Adult MeSH
- Carrier State * microbiology epidemiology MeSH
- Prevalence MeSH
- Staphylococcal Infections * microbiology epidemiology MeSH
- Staphylococcus aureus * genetics isolation & purification drug effects classification MeSH
- Students, Medical * MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
The emergence of biofilm-induced drug tolerance poses a critical challenge to public healthcare management. Pseudomonas aeruginosa, a gram-negative opportunistic bacterium, is involved in various biofilm-associated infections in human hosts. Towards this direction, in the present study, a combinatorial approach has been explored as it is a demonstrably effective strategy for managing microbial infections. Thus, P. aeruginosa has been treated with cuminaldehyde (a naturally occurring phytochemical) and gentamicin (an aminoglycoside antibiotic) in connection to the effective management of the biofilm challenges. It was also observed that the test molecules could show increased antimicrobial activity against P. aeruginosa. A fractional inhibitory concentration index (FICI) of 0.65 suggested an additive interaction between cuminaldehyde and gentamicin. Besides, a series of experiments such as crystal violet assay, estimation of extracellular polymeric substance (EPS), and microscopic images indicated that an enhanced antibiofilm activity was obtained when the selected compounds were applied together on P. aeruginosa. Furthermore, the combination of the selected compounds was found to reduce the secretion of virulence factors from P. aeruginosa. Taken together, this study suggested that the combinatorial application of cuminaldehyde and gentamicin could be considered an effective approach towards the control of biofilm-linked infections caused by P. aeruginosa.
- MeSH
- Anti-Bacterial Agents * pharmacology MeSH
- Benzaldehydes * pharmacology MeSH
- Biofilms * drug effects MeSH
- Cymenes pharmacology MeSH
- Virulence Factors MeSH
- Gentamicins * pharmacology MeSH
- Humans MeSH
- Microbial Sensitivity Tests * MeSH
- Pseudomonas aeruginosa * drug effects physiology MeSH
- Drug Synergism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Environmental pollution is a serious problem that can cause sicknesses, fatality, and biological contaminants such as bacteria, which can trigger allergic reactions and infectious illnesses. There is also evidence that environmental pollutants can have an impact on the gut microbiome and contribute to the development of various mental health and metabolic disorders. This study aimed to study the antibiotic resistance and virulence potential of environmental Pseudomonas aeruginosa (P. aeruginosa) isolates in slaughterhouses. A total of 100 samples were collected from different slaughterhouse tools. The samples were identified by cultural and biochemical tests and confirmed by the VITEK 2 system. P. aeruginosa isolates were further confirmed by CHROMagarTM Pseudomonas and genetically by rpsL gene analysis. Molecular screening of virulence genes (fimH, papC, lasB, rhlI, lasI, csgA, toxA, and hly) and antibiotic resistance genes (blaCTX-M, blaAmpC, blaSHV, blaNDM, IMP-1, aac(6')-Ib-, ant(4')IIb, mexY, TEM, tetA, and qnrB) by PCR and testing the antibiotic sensitivity, biofilm formation, and production of pigments, and hemolysin were carried out in all isolated strains. A total of 62 isolates were identified as P. aeruginosa. All P. aeruginosa isolates were multidrug-resistant and most of them have multiple resistant genes. blaCTX-M gene was detected in all strains; 23 (37.1%) strains have the ability for biofilm formation, 33 strains had virulence genes, and 26 isolates from them have more than one virulence genes. There should be probably 60 (96.8%) P. aeruginosa strains that produce pyocyanin pigment. Slaughterhouse tools are sources for multidrug-resistant and virulent pathogenic microorganisms which are a serious health problem. Low-hygienic slaughterhouses could be a reservoir for resistance and virulence genes which could then be transferred to other pathogens.
- MeSH
- Anti-Bacterial Agents * pharmacology MeSH
- Drug Resistance, Bacterial genetics MeSH
- Biofilms drug effects growth & development MeSH
- Virulence Factors * genetics MeSH
- Abattoirs * MeSH
- Microbial Sensitivity Tests * MeSH
- Environmental Microbiology MeSH
- Pseudomonas aeruginosa * genetics drug effects pathogenicity isolation & purification MeSH
- Virulence genetics MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
Bacillus thuringiensis (Bt) is known for its Cry and Vip3A pesticidal proteins with high selectivity to target pests. Here, we assessed the potential of a novel neotropical Bt strain (UFT038) against six lepidopteran pests, including two Cry-resistant populations of fall armyworm, Spodoptera frugiperda. We also sequenced and analyzed the genome of Bt UFT038 to identify genes involved in insecticidal activities or encoding other virulence factors. In toxicological bioassays, Bt UFT038 killed and inhibited the neonate growth in a concentration-dependent manner. Bt UFT038 and HD-1 were equally toxic against S. cosmioides, S. frugiperda (S_Bt and R_Cry1 + 2Ab populations), Helicoverpa zea, and H. armigera. However, larval growth inhibition results indicated that Bt UFT038 was more toxic than HD-1 to S. cosmioides, while HD-1 was more active against Chrysodeixis includens. The draft genome of Bt UFT038 showed the cry1Aa8, cry1Ac11, cry1Ia44, cry2Aa9, cry2Ab35, and vip3Af5 genes. Besides this, genes encoding the virulence factors (inhA, plcA, piplC, sph, and chi1-2) and toxins (alo, cytK, hlyIII, hblA-D, and nheA-C) were also identified. Collectively, our findings reveal the potential of the Bt UFT038 strain as a source of insecticidal genes against lepidopteran pests, including S. cosmioides and S. frugiperda.
- MeSH
- Bacillus thuringiensis * genetics metabolism MeSH
- Bacterial Proteins genetics metabolism MeSH
- Pest Control, Biological MeSH
- Endotoxins metabolism MeSH
- Virulence Factors metabolism MeSH
- Glycine max MeSH
- Hemolysin Proteins genetics metabolism pharmacology MeSH
- Insecticides * pharmacology metabolism MeSH
- Larva MeSH
- Humans MeSH
- Moths * MeSH
- Infant, Newborn MeSH
- Spodoptera metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Infant, Newborn MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
BACKGROUND: Multidrug-resistant (MDR) bacteria pose a significant challenge to the treatment of infectious diseases. Of particular concern are members of the Klebsiella pneumoniae species complex (KpSC), which are frequently associated with hospital-acquired infections and have the potential to spread outside hospitals via wastewaters. In this study, we aimed to investigate the occurrence and phylogenetic relatedness of MDR KpSC from patients with urinary tract infections (UTIs), hospital sewage, municipal wastewater treatment plants (mWWTPs) and surface waters and to evaluate the clinical relevance of the KpSC subspecies. METHODS: A total of 372 KpSC isolates resistant to third-generation cephalosporins and/or meropenem were collected from patients (n = 130), hospital sewage (n = 95), inflow (n = 54) and outflow from the mWWTPs (n = 63), river upstream (n = 13) and downstream mWWTPs (n = 17) from three cities in the Czech Republic. The isolates were characterized by antimicrobial susceptibility testing and whole-genome sequencing (Illumina). The presence of antibiotic resistance genes, plasmid replicons and virulence-associated factors was determined. A phylogenetic tree and single nucleotide polymorphism matrix were created to reveal the relatedness between isolates. RESULTS: The presence of MDR KpSC isolates (95%) was identified in all water sources and locations. Most isolates (99.7%) produced extended-spectrum beta-lactamases encoded by blaCTX-M-15. Resistance to carbapenems (5%) was observed mostly in wastewaters, but carbapenemase genes, such as blaGES-51 (n = 10), blaOXA-48 (n = 4), blaNDM-1 (n = 4) and blaKPC-3 (n = 1), were found in isolates from all tested locations and different sources except rivers. Among the 73 different sequence types (STs), phylogenetically related isolates were observed only among the ST307 lineage. Phylogenetic analysis revealed the transmission of this lineage from patients to the mWWTP and from the mWWTP to the adjacent river and the presence of the ST307 clone in the mWWTP over eight months. We confirmed the frequent abundance of K. pneumoniae (K. pneumoniae sensu stricto and K. pneumoniae subsp. ozaenae) in patients suffering from UTIs. K. variicola isolates formed only a minor proportion of UTIs, and K. quasipneumoniae was not found among UTIs isolates; however, these subspecies were frequently observed in hospital sewage communities during the first sampling period. CONCLUSION: This study provides evidence of the transmission and persistence of the ST307 lineage from UTIs isolates via mWWTPs to surface waters. Isolates from UTIs consisted mostly of K. pneumoniae. Other isolates of KpSC were observed in hospital wastewaters, which implies the impact of sources other than UTIs. This study highlights the influence of urban wastewaters on the spread of MDR KpSC to receiving environments.
- MeSH
- Anti-Bacterial Agents * pharmacology MeSH
- Bacterial Proteins genetics MeSH
- beta-Lactamases * genetics MeSH
- Phylogeny * MeSH
- Klebsiella Infections * microbiology epidemiology MeSH
- Urinary Tract Infections microbiology epidemiology MeSH
- Cross Infection microbiology epidemiology MeSH
- Klebsiella pneumoniae * drug effects genetics isolation & purification classification MeSH
- Humans MeSH
- Microbial Sensitivity Tests * MeSH
- Drug Resistance, Multiple, Bacterial * MeSH
- Hospitals * MeSH
- Wastewater * microbiology MeSH
- Sewage microbiology MeSH
- Whole Genome Sequencing MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
Bordetella pertussis is the causative agent of whooping cough in humans, a disease that has recently experienced a resurgence. In contrast, Bordetella bronchiseptica infects the respiratory tract of various mammalian species, causing a range of symptoms from asymptomatic chronic carriage to acute illness. Both pathogens utilize type III secretion system (T3SS) to deliver the effector protein BteA into host cells. Once injected, BteA triggers a cascade of events leading to caspase 1-independent necrosis through a mechanism that remains incompletely understood. We demonstrate that BteA-induced cell death is characterized by the fragmentation of the cellular endoplasmic reticulum and mitochondria, the formation of necrotic balloon-like protrusions, and plasma membrane permeabilization. Importantly, genome-wide CRISPR-Cas9 screen targeting 19,050 genes failed to identify any host factors required for BteA cytotoxicity, suggesting that BteA does not require a single nonessential host factor for its cytotoxicity. We further reveal that BteA triggers a rapid and sustained influx of calcium ions, which is associated with organelle fragmentation and plasma membrane permeabilization. The sustained elevation of cytosolic Ca2+ levels results in mitochondrial calcium overload, mitochondrial swelling, cristolysis, and loss of mitochondrial membrane potential. Inhibition of calcium channels with 2-APB delays both the Ca2+ influx and BteA-induced cell death. Our findings indicate that BteA exploits essential host processes and/or redundant pathways to disrupt calcium homeostasis and mitochondrial function, ultimately leading to host cell death.IMPORTANCEThe respiratory pathogens Bordetella pertussis and Bordetella bronchiseptica exhibit cytotoxicity toward a variety of mammalian cells, which depends on the type III secretion effector BteA. Moreover, the increased virulence of B. bronchiseptica is associated with enhanced expression of T3SS and BteA. However, the molecular mechanism underlying BteA cytotoxicity is elusive. In this study, we performed a CRISPR-Cas9 screen, revealing that BteA-induced cell death depends on essential or redundant host processes. Additionally, we demonstrate that BteA disrupts calcium homeostasis, which leads to mitochondrial dysfunction and cell death. These findings contribute to closing the gap in our understanding of the signaling cascades targeted by BteA.
- MeSH
- Bacterial Proteins * metabolism genetics MeSH
- Bordetella bronchiseptica genetics metabolism drug effects MeSH
- Bordetella pertussis genetics pathogenicity metabolism drug effects MeSH
- Cell Death * drug effects MeSH
- Endoplasmic Reticulum metabolism drug effects MeSH
- Homeostasis * MeSH
- Host-Pathogen Interactions MeSH
- Humans MeSH
- Mitochondria metabolism drug effects MeSH
- Type III Secretion Systems metabolism genetics MeSH
- Calcium * metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Gram-positive bacteria are responsible for a wide range of infections in humans. In most Gram-positive bacteria, sortase A plays a significant role in attaching virulence factors to the bacteria's cell wall. These cell surface proteins play a significant role in virulence and pathogenesis. Even though antibiotics are available to treat these infections, there is a continuous search for an alternative strategy due to an increase in antibiotic resistance. Thus, using anti-sortase drugs to combat these bacterial infections may be a promising approach. Here, we describe a method for targeting Gram-positive bacterial infection by combining curcumin and trans-chalcone as sortase A inhibitors. We have used curcumin and trans-chalcone alone and in combination as a sortase A inhibitor. We have seen ~78%, ~43%, and ~94% inhibition when treated with curcumin, trans-chalcone, and a combination of both compounds, respectively. The compounds have also shown a significant effect on biofilm formation, IgG binding, protein A recruitment, and IgG deposition. We discovered that combining curcumin and trans-chalcone is more effective against Gram-positive bacteria than either compound alone. The present work demonstrated that a combination of these natural compounds could be used as an antivirulence therapy against Gram-positive bacterial infection.
- MeSH
- Aminoacyltransferases * antagonists & inhibitors metabolism MeSH
- Anti-Bacterial Agents * pharmacology chemistry MeSH
- Bacterial Proteins * metabolism antagonists & inhibitors MeSH
- Biofilms * drug effects MeSH
- Chalcone * pharmacology chemistry MeSH
- Cysteine Endopeptidases * metabolism MeSH
- Virulence Factors metabolism MeSH
- Gram-Positive Bacterial Infections drug therapy microbiology MeSH
- Gram-Positive Bacteria drug effects MeSH
- Curcumin * pharmacology chemistry MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Virulence drug effects MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Mycoviruses are viruses that infect fungi and are widespread across all major fungal taxa, exhibiting great biological diversity. Since their discovery in the 1960s, researchers have observed a myriad of fungal phenotypes altered due to mycoviral infection. In this review, we examine the nuanced world of mycoviruses in the context of the medically and agriculturally important fungal genus, Aspergillus. The advent of RNA sequencing has revealed a previous underestimate of viral prevalence in fungi, in particular linear single-stranded RNA viruses, and here we outline the diverse viral families known to date that contain mycoviruses infecting Aspergillus. Furthermore, we describe these novel mycoviruses, highlighting those with peculiar genome structures, such as a split RNA dependent RNA polymerase gene. Next, we delineate notable mycovirus-mediated phenotypes in Aspergillus, in particular reporting on observations of mycoviruses that affect their fungal host's virulence and explore how this may relate to virus-mediated decreased stress tolerance. Furthermore, mycovirus effects on microbial competition and antifungal resistance are discussed. The factors that influence the manifestation of these phenotypes, such as temperature, fungal life stage, and infection with multiple viruses, among others, are also evaluated. In addition, we attempt to elucidate the molecular mechanisms that underpin these phenotypes, examining how mycoviruses can be targets, triggers, and even suppressors of RNA silencing and how this can affect fungal gene expression and phenotypes. Finally, we highlight the potential therapeutic applications of mycoviruses and how, in an approach analogous to bacteriophage therapy, their ability to produce hypovirulence in Aspergillus might be used to attenuate invasive aspergillosis infections in humans.
- Publication type
- Journal Article MeSH
- Review MeSH
