High oral carriage of multidrug resistant Gram-negative bacilli in adolescents: the SOPKARD-Junior study.

Introduction: The colonization of the oral cavity by potentially pathogenic antimicrobial-resistant bacteria in adolescents and its consequences is very poorly understood. The present study focused on the occurrence of oral colonization by Gram-negative bacilli (GNB) and their multidrug resistance, including the production of extended-spectrum ß-lactamases (ESBLs) and carbapenemases, among healthy adolescents and risk factors associated with GNB colonization. Materials and methods: This study was conducted as part of "A program for the early detection of risk factors for lifestyle diseases SOPKARD-Junior" (SOPKARD-Junior). Oral samples were collected from 182 adolescents from four public elementary schools in Sopot, Poland, aged 13-14 years. Bacterial strains were identified by the MALDI-TOF MS method. Screening of antimicrobial resistance was performed using a disk diffusion method. The NG-Test® CARBA-5 was used to detect and differentiate the five most widely distributed carbapenemases. Demographic and clinical data were collected and statistical analysis of risk factors was performed. Results: A total of 68 out of 182 (37.4%) healthy adolescents was documented oral colonization with Gram-negative bacilli, including 50/182 (27.5%) multidrug resistant (MDR-GNB) strains. Over 60% of oral carriage concerned three main genera Enterobacter spp., Pseudomonas spp., and Serratia spp., which were detected in 22.1%, 19.1%, and 19.1% of participants, respectively. Citrobacter spp., Escherichia coli, Klebsiella spp., Hafnia spp., Aeromonas spp., Acinetobacter spp., and Stenotrophomonas spp. were also isolated. The antimicrobial resistance to ampicillin (100%), ceftazidime (69.1%), meropenem (60.3%), gentamycin (60.3%), piperacillin/tazobactam (52.9%), and piperacillin (45.6%) were the most common. Among 73.5% GNB strains multidrug resistance was observed, including all Pseudomonas spp. strains. Among MDR-GNB, 30.4% were resistant to four groups of antibiotics, half of the MDR Pseudomonas spp. strains were resistant to 10 groups of antibiotics. Extended-spectrum ß-lactamases were produced by Enterobacter cloacae, Klebsiella spp., and Serratia spp. (7.4%). Colonization by ESBLs-positive GNB strains was significantly associated with recurrent respiratory infections, nasal congestion, and bronchitis (p<0.05). Conclusion: Our study revealed high oral carriage of multi-drug resistant Gram-negative bacilli in healthy adolescents and the association of ESBL-producing strains with respiratory infections. Further studies on oral colonization with GNB are necessary due to the possibility of distinct infections and the acquisition of antibiotic resistance by resident microbiota.

Antioxidant and antimicrobial properties of an extract rich in proteins obtained from Trametes versicolor.

Introduction: Bioactive proteins and peptides generated from fruit, vegetables, meat or fish have great potential as functional food or substitutes for antibiotics. In recent years it has also been demonstrated that the fungus kingdom could be a source of these compounds. The study investigated the bioactivity of an extract of the lignicolous fungus Trametes versicolor and its hydrolysate. Material and Methods: The fungus was collected in a mixed forest in October, extracted and hydrolysed. To inspect the protein and peptide profiles before and after hydrolysis, matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometric analysis was performed. To evaluate the antioxidant properties of the preparations, 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+) radical scavenging assays were used. The activity of the fungus extract and hydrolysate against Aeromonas veronii, Bacillus cereus, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella Typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis was determined by the minimum inhibitory concentration and minimum bactericidal concentration values. Results: The extract and its hydrolysate showed almost 100% ABTS•+ and DPPH• radical scavenging with a low half maximal inhibitory concentration. The water extract and hydrolysate of T. versicolor exhibited antimicrobial activity against two S. aureus strains, E. coli, P. aeruginosa and Salmonella Typhimurium. Conclusion: These results provide compelling evidence that the analysed fungus extract and its hydrolysate hold promise with their antibacterial and antioxidant properties.

Recent Trends in the Application of Oilseed-Derived Protein Hydrolysates as Functional Foods.

Oilseed-derived proteins have emerged as an excellent alternative to animal sources for the production of bioactive peptides. The bioactivities exhibited by peptides derived from plant proteins encompass a wide range of health-promoting and disease-preventing effects. Peptides demonstrate potential capabilities in managing diseases associated with free radicals and regulating blood pressure. They can also exhibit properties that lower blood sugar levels and modify immune responses. In addition to their bioactivities, plant-derived bioactive peptides also possess various functional properties that contribute to their versatility. An illustration of this potential can be the ability of peptides to significantly improve food preservation and reduce lipid content. Consequently, plant-derived bioactive peptides hold great promise as ingredients to develop functional products. This comprehensive review aims to provide an overview of the research progress made in the elucidation of the biological activities and functional properties of oilseed-derived proteins. The ultimate objective is to enhance the understanding of plant-derived bioactive peptides and provide valuable insights for further research and use in the food and medicine industries.

Egg Yolk as a New Source of Peptides with Antioxidant and Antimicrobial Properties.

A significant increase in interest in food-derived peptides obtained mostly through enzymatic reactions has been observed in the past few years. One of the best sources of bioactive peptides are defatted egg yolk proteins, which can potentially find application as high-quality nutritional supplements for infants with cow's milk protein intolerance and as natural preservatives. The aim of this study was to obtain peptides from defatted egg yolk protein, to study their antioxidant and antimicrobial properties, and to identify peptides with bioactive properties To control the course of the process, MALDI-TOF/MS (matrix-assisted laser desorption/ionization time-of flight/mass spectrometry) spectra were also examined. The peptide mixture obtained through enzyme digestion was tested for its antioxidant properties by measuring the scavenging activity in 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization (ABTS•+), and ferric reducing activity (FRAP) assays. Antimicrobial activity was also studied. The peptide mixture exhibited significant antioxidant activity: DPPH-1776.66 ± 32.99, ABTS-390.43 ± 8.92, and FRAP-16.45 ± 0.19. The inhibition of bacterial growth by two concentrations of the peptide mixture was examined. The best result was obtained in Bacillus cereus, with an inhibition zone of 20.0 ± 1.0 and 10.7 ± 0.6 mm at the concentrations of 50 and 25 mg/mL, respectively. The results of the study suggest that the mixture of egg yolk peptides may exhibit a number of health-promoting properties.

Indole-Acrylonitrile Derivatives as Potential Antitumor and Antimicrobial Agents-Synthesis, In Vitro and In Silico Studies.

A series of 2-(1H-indol-2-yl)-3-acrylonitrile derivatives, 2a-x, 3, 4a-b, 5a-d, 6a-b, and 7, were synthesized as potential antitumor and antimicrobial agents. The structures of the prepared compounds were evaluated based on elemental analysis, IR, 1H- and 13NMR, as well as MS spectra. X-ray crystal analysis of the representative 2-(1H-indol-2-yl)-3-acrylonitrile 2l showed that the acrylonitrile double bond was Z-configured. All compounds were screened at the National Cancer Institute (USA) for their activities against a panel of approximately 60 human tumor cell lines and the relationship between structure and in vitro antitumor activity is discussed. Compounds of interest 2l and 5a-d showed significant growth inhibition potency against various tumor cell lines with the mean midpoint GI50 values of all tests in the range of 0.38-7.91 µM. The prominent compound with remarkable activity (GI50 = 0.0244-5.06 µM) and high potency (TGI = 0.0866-0.938 µM) against some cell lines of leukemia (HL-60(TB)), non-small cell lung cancer (NCI-H522), colon cancer (COLO 205), CNS cancer (SF-539, SNB-75), ovarian cancer ((OVCAR-3), renal cancer (A498, RXF 393), and breast cancer (MDA-MB-468) was 3-[4-(dimethylamino)phenyl]-2-(1-methyl-1H-indol-2-yl)acrylonitrile (5c). Moreover, the selected 2-(1H-indol-2-yl)-3-acrylonitriles 2a-c and 2e-x were evaluated for their antibacterial and antifungal activities against Gram-positive and Gram-negative pathogens as well as Candida albicans. Among them, 2-(1H-indol-2-yl)-3-(1H-pyrrol-2-yl)acrylonitrile (2x) showed the most potent antimicrobial activity and therefore it can be considered as a lead structure for further development of antimicrobial agents. Finally, molecular docking studies as well as drug-likeness and ADME profile prediction were carried out.

Effectiveness of a 12-Month Online Weight Reduction Program in Cohorts with Different Baseline BMI-A Prospective Cohort Study.

The purpose of this article was to answer the question of whether people who want to reduce their body weight can achieve different results depending on their baseline BMI and whether the rate of weight loss is constant over the months of intervention. The study included 400 individuals aged 19 to 55 years with a mean BMI of 31.83 ± 4.77 (min 25.1 max 51.8). Men comprised 190 subjects and women 210 subjects. The participants were divided into three groups with the following BMI: overweight, class 1 obesity, and obesity class > 1 (class 2 and 3 combined). BMI groups were randomized by gender, the number of trainings per week, training time, intervention length, and intervention type. The online intervention consisted of a 15% energy deficit diet and training. Over the 12-month dietary intervention, overweight subjects reduced average body weight by 16.6%. The group with class 1 obesity reduced body weight by 15.7%. The group with obesity class > 1 reduced mean body weight by 15.4%. The relative weight reduction in the overweight group was significantly greater than in the other obesity groups (p = 0.007). In all groups, the rate of weight loss from month-to-month was statistically significant (p = 0.0001), ranging between 0.6 and 2.6% per month. The results indicate that overweight individuals are likely to experience a percent greater weight loss as a result of a comprehensive lifestyle intervention. Regardless of baseline BMI, the observed weight loss was consistent from month-to-month throughout the 12-month period, which may indicate that the diet, as well as, the training plan were properly tailored to the subjects' needs and that they were highly motivated to participate in the program throughout its course. Properly conducted lifestyle intervention enables significant weight loss regardless of baseline BMI values.

Protein Hydrolysates Derived from Animals and Plants-A Review of Production Methods and Antioxidant Activity.

There is currently considerable interest on the use of animal, plant, and fungal sources in the production of bioactive peptides, as evidenced by the substantial body of research on the topic. Such sources provide cheap and environmentally friendly material as it often includes waste and by-products. Enzymatic hydrolysis is considered an efficient method of obtaining peptides capable of antioxidant activity. Those properties have been proven in terms of radical-scavenging capacity using the DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2-azinobis-(3-ethyl-benzothiazoline-6-sulphonic acid)), hydroxyl and superoxide radical methods. Additionally, the reducing power, ferrous ion-chelating (FIC), ferric reducing antioxidant power (FRAP), and the ability of the protein hydrolysates to inhibit lipid peroxidation have also been explored. The results collected in this review clearly indicate that the substrate properties, as well as the conditions under which the hydrolysis reaction is carried out, affect the final antioxidant potential of the obtained peptides. This is mainly due to the structural properties of the obtained compounds such as size or amino acid sequences.

Alternative and online weight loss interventions during Covid-19 pandemic lockdown.

The problem of obesity is affecting an increasing number of people worldwide. The COVID-19 pandemic and the required social distancing, which make it impossible to see a dietitian, present new challenges and require the development of new ways of working with overweight individuals. Based on research and practice, dietetic services are beginning to shift from in-office consultations to a form of online consultation using mobile apps and websites. This literature review aims to critically analyze the scientific evidence for the effectiveness of interventions targeting weight loss in overweight or obese individuals focusing on behavioral and online interventions. The data from the presented studies suggest that the effectiveness of online interventions to control body weight is high enough to be used in nutritional education and in weight reduction or maintenance. The main advantage is a wide access to them for the public.

Microbiological, Clinical and Radiological Aspects of Diabetic Foot Ulcers Infected with Methicillin-Resistant and -Sensitive Staphylococcus aureus.

Diabetic foot ulcer (DFU) is one of the most common chronic complications of diabetes. This study aimed to assess the factors with an impact on the infection of diabetic foot ulcers by methicillin-resistant S. aureus and to evaluate the influence of methicillin resistance on the frequency of osteitis (based on classic X-ray images). A total of 863 patients suffering from DFU were analyzed during the study period. Out of 201 isolated S. aureus cases, 31 (15.4%) were methicillin-resistant (MRSA). MRSA infections were associated with a higher incidence of osteitis compared to MSSA infections (p << 0.0001), both the occurrence of smaller (<50%)) and greater (>50%) inflammatory bone changes (p << 0.0001). Furthermore, MRSA occurred significantly more frequently in men than in women (p < 0.01) and more often among patients with type 2 diabetes than among patients with type 1 diabetes (p < 0.05). MRSA were isolated statistically less often in overweight patients than in patients with normal BMI (p < 0.05). DFUs infected with MRSA were significantly more frequently associated with the presence of Pseudomonas sp. and other non-fermenting bacilli than those infected with MSSA (p < 0.05). To conclude, osteitis incidence is related to MRSA infection in patients with diabetic foot ulcers; thus, patients infected by S. aureus should be closely monitored in the course of using antibiotics and treated with narrow-spectrum antibiotics.

Anticancer activity of lactic acid bacteria.

Lactic acid bacteria (LAB), a group of Gram-positive microorganisms naturally occurring in fermented food products and used as probiotics, have been gaining the interest of researchers for years. LAB are potent, albeit still not wholly understood, source of bioactive compounds with various functions and activity. Metabolites of LAB, among others, short-chain fatty acids, exopolysaccharides and bacteriocins have promising anticancer potential. Research on the interactions between the bioactive metabolites of LAB and immune mechanisms demonstrated that these substances could exert a strong immunomodulatory effect, which would explain their vast therapeutic potential. The anticancer activity of LAB was confirmed both in vitro and in animal models against cancer cells from various malignancies. LAB inhibit tumor growth through various mechanisms, including antiproliferative activity, induction of apoptosis, cell cycle arrest, as well as through antimutagenic, antiangiogenic and anti-inflammatory effects. The aim of this review was to summarize the most recent data about the anticancer activity of LAB, with particular emphasis on the most promising bioactive compounds with potential clinical application.

Probiotics: Should All Patients Take Them?

The usefulness of probiotics in the treatment as well as prevention of many infections and disorders has been confirmed by previous clinical studies. They can protect not only against gastrointestinal diseases such as diarrhea or enteritis but they have proven efficacy against pneumonia, urogenital infection, depression/anxiety, cancer metastasis, obesity, and others. However, it should be mentioned that not all clinical trials have shown improvement of health in patients undergoing probiotic treatment, and very rarely have even reported that probiotic strains may be the causative agents of opportunistic infections. Studies have documented cases of sepsis/bacteremia, endocarditis, liver abscess, pneumonia, and fungemia caused by probiotic strains, mainly in high-risk groups. This review summarizes the cases of infections caused by probiotic strains and the potential hazard associated with the supplementation of probiotics in seriously ill and hospitalized patients.

Staphylococcus aureus Isolated from the Oral Cavity: Phage Susceptibility in Relation to Antibiotic Resistance.

Nowadays, research on bacteriophage therapy and its potential use in combination with antibiotics has been gaining momentum. One hundred and ten oral Staphylococcus aureus isolates were phage-typed and their antibiotic resistance was determined by standard and molecular methods. The prevalence of MSSA and MRSA strains was 89.1% and 10.9%, respectively. Nearly all (91.8%) analyzed isolates, whether MSSA or MRSA, were susceptible to the phages used from the international set. The highest lytic activity showed phages 79 and 52 A from lytic group I. The predominant phage groups were mixed, the I+III group and a mixed group containing phages from at least three various lytic groups. S. aureus strains sensitive to phage group I were usually resistant to penicillin and susceptible to ciprofloxacin, whereas the strains typeable with group V or group V with the 95 phage were susceptible to most antibiotics. Epidemic CA-MRSA strains (SCCmecIV) of phage type 80/81 carried Panton-Valentine leucocidin genes. Considering the high sensitivity of oral S. aureus to the analyzed phages and the promising results of phage therapies reported by other authors, phage cocktails or phage-antibiotic combinations may potentially find applications in both the prevention and eradication of staphylococcal infections.

Distribution and antibiotic-resistance of different Staphylococcus species identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) isolated from the oral cavity.

BACKGROUND: The use of antibiotics in dentistry is associated with the emergence and spread of antibiotic-resistant microorganisms, including commensal staphylococci. METHODS: A total of 367 oral samples were collected, from which staphylococci were isolated and identified by using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF). The antibiotic susceptibility of the isolates was determined and molecular characteristics for methicillin-resistant staphylococci was performed. RESULTS: A total of 103 coagulase-negative staphylococci (CoNS), among them S. warneri, S. haemolyticus, S. saprophyticus, S. pasteuri, S. epidermidis, S. hominis, S. xylosus, S. equorum, S. kloosii, S. succinus, S. cohnii, and S. simulans, were confirmed by MALDI-TOF. Resistance to most tested antibiotics was statistically higher in CoNS than in S. aureus isolates (P-value < 0.05). CoNS isolates showed high resistance to penicillin (S. saprophyticus 88.9%), erythromycin (S. haemolyticus 84.6%), fusidic acid (S. saprophyticus 77.8%), co-trimoxazole (S. epidermidis 71.4%), gentamicin (S. warneri 63.8%), and tetracycline (S. saprophyticus 55.6%). Multidrug resistance was largely observed, especially among S. haemolyticus and S. saprophyticus species. Methicillin-resistance in S. haemolyticus (38.5%), S. saprophyticus (22.2%) and S. aureus (13.5%) was associated with the presence of the mecA gene and SCCmec type IV or V. CONCLUSION: Coagulase-negative staphylococci, especially S. haemolyticus and S. saprophyticus, seem to be a reservoir of methicillin resistance and multidrug resistance in the oral cavity.

COVID-19 in dental care: What do we know?

SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) is primarily transmitted by airborne droplets and its spread is favored by close human contact, thus the COVID-19 pandemic is the new challenge in dental practice. The oral cavity was considered as a SARS-CoV-2 reservoir, the viruses were detected in the saliva and periodontal pockets of infected persons. Therefore, aside from the most common symptoms, COVID-19 can manifest as lesions in the oral cavity. Due to the high risk of cross-contamination in the dental office, new precautionary measures were implemented in professional dental care to ensure safety for both, dental staff and patients. Given the dynamically changing situation, dental practitioners should follow local guidelines and implement them according to current needs and available resources. The key to success is to reduce the risk of cross-infection with SARS-CoV-2 at no cost to the good oral health of the population.

Novel staphylococci nucH taxonomical marker used in identification of human-associated Staphylococcus succinus subsp. casei.

The aim of our study was to assess the sequencing of unique nucH gene fragment based on performed bioinformatics analysis as a novel diagnostic method for the identification of difficult to identify staphylococcal human pathogenic strains. Initially, PCR-RFLP-rrn analysis specific to the spacers between 16SrDNA and 23SrDNA followed by HhaI restriction analysis was performed. Further, sequencing of nucH and 16S rDNA genes fragments was carried out. Blast analysis from the NCBI showed 99% similarity of nucH gene fragment with reference genomic DNA for S. succinus with the accession no. CP018199. This result was also confirmed by MALDI-TOF analysis. Sequencing analysis of 16S rDNA gene fragment allowed for 100% identification of two strains isolated from human samples as Staphylococus succinus subsp. casei. Sequencing of identified unique nucH gene fragment seems to be a promising diagnostic assay for the identification of Staphylococcus species. Based on our results, we can assume that probably other Staphylococcus species originated from different clinical samples could be identified using nucH gene sequencing method we developed. However, an extension of the genetic databases with a substantially bigger number of reference staphylococcal species for nucH gene is needed to make this method better than widely used standard 16S rDNA sequencing assay. To the best of our knowledge, it is the second published isolation of S. succinus subsp. casei from human clinical specimens. Moreover, possibility of decreasing the number of dimensions from multi-PCR-bands results using ribotyping analysis is also described.

Presence of egc-positive major clones ST 45, 30 and 22 among methicillin-resistant and methicillin-susceptible oral Staphylococcus aureus strains.

The oral cavity may comprise a significant reservoir for Staphylococcus aureus but the data on molecular epidemiology and clonal distribution of oral strains are really scarce. This study aimed to evaluate the clonal relatedness in S. aureus isolated from oral cavity and their relationship with carriage of virulence genes, and antimicrobial resistance profiles. A total of 139 oral S. aureus isolates were obtained from 2327 analysed oral samples of dental patients. Antimicrobial susceptibility testing was performed. Isolates were characterized using protein A gene (spa) typing, spa-CC clonal complexes, toxin genes and SCCmec typing for MRSA. High resistance rates for penicillin, tetracycline and gentamicin were detected, respectively 58.3%, 42.4%, and 35.2%. Twelve (8.6%) S. aureus isolates were identified as MRSA. All of MRSA isolates were mecA-positive and mecC-negative. SCCmec IV was the most common type (66.7%), which was typical for community-acquired MRSA (CA-MRSA). Overall, the enterotoxin gene cluster (egc) was the most frequent detected virulence factor (44.9%), both in MSSA and MRSA isolates. Presence of genes encoding for the enterotoxins (sea, seb, sec, seh, sek), exfoliative toxin A (eta), and toxic shock syndrome toxin-1 (tst) was also observed. Strains carrying lukS-PV/lukF-PV genes belonged to SCCmecV- spa type t437. The most prevalent spa types were t091, t015, t084, t002, t571, and t026 among all 57 identified. Spa types, including 3 new ones, grouped in 6 different spa-CC clonal complexes, with four major dominated; CC45, CC30, CC5, and CC15. This study demonstrated that both methicillin-susceptible and methicillin-resistant major European clones of S. aureus could be isolated from the oral cavity of dental patients, with the emergence of PVL-positive CA-MRSA strains. The oral cavity should be considered as a possible source of toxigenic egc-positive S. aureus strains, in terms of potential risk of cross-infection and dissemination to other body sites.

In Vitro Efficiency of Antimicrobial Peptides against Staphylococcal Pathogens Associated with Canine Pyoderma.

The emergence of staphylococcal canine pathogens resistant to multiple antimicrobial agents is a growing and urgent problem in veterinary practice. Antimicrobial peptides (AMPs) seem to be a promising alternative to conventional antibiotics. The aim of this in vitro study was to evaluate the antimicrobial activity of selected AMPs against pathogenic staphylococcal strains, including multidrug- and methicillin-resistant strains isolated from canine pyoderma cases. Seven antimicrobial peptides (aurein 1.2, CAMEL, citropin 1.1, protegrin-1, pexiganan, temporin A and uperin 3.6) synthesized by the 9-fluorenylmethoxycarbonyl (Fmoc) solid-phase method were tested. The minimal inhibitory and minimal bactericidal concentrations (MIC and MBC) were determined by the broth microdilution method. The study showed that analyzed AMPs exerted an extensive effect against canine pathogens, with the most active peptide being uperin 3.6. The tested AMPs were equally efficient against both resistant- and susceptible staphylococcal strains and were more efficient against Staphylococcus pseudintermedius than against Staphylococcus aureus strains. Our findings are particularly interesting from a clinical perspective, as they point to AMPs as potential therapeutic topical agents in canine pyoderma cases associated with antimicrobial resistance of staphylococci.

Genetic diversity and antimicrobial resistance of Staphylococcus aureus from recurrent tonsillitis in children.

The aim of this study was to analyze the prevalence of Staphylococcus aureus in the tonsils of children subjected tonsillectomy due to recurrent tonsilitis and to determine the spa types of the pathogens, carriage of virulence genes and antimicrobial resistance profiles. The study included 73 tonsillectomized children. Bacteria, including S. aureus were isolated from tonsillar surface prior to tonsillectomy, recovered from tonsillar core at the time of the surgery, and from posterior pharynx 2-4 weeks after the procedure. Staphylococcus aureus isolates were compared by spa typing, tested for antimicrobial susceptibility and for the presence of superantigenic toxin genes (sea-seu, eta, etb, tst, lukS/lukF-PV) by multiplex polymerase chain reaction. Seventy-three patients (mean 7.1 ± 4.1 years, 61.6% male) were assessed. The most commonly isolated bacteria were S. aureus. The largest proportion of staphylococcal isolates originated from tonsillar core (63%), followed by tonsillar surface (45.1%) and posterior pharynx in tonsillectomized children (18.2%, p = 0.007). Five (6.3%) isolates were identified as MRSA (mecA-positive). Up to 67.5% of the isolates synthesized penicillinases (blaZ-positive isolates), and 8.8% displayed MLSB resistance. The superantigenic toxin genes were detected in more than half of examined isolates (56.3%). spa types t091, t084, and t002, and clonal complexes (CCs) CC7, CC45, and CC30 turned out to be most common. Staphylococcus aureus associated with RT in children showed pathogenicity potential and considerable genetic diversity, and no clones were found to be specific for this condition although further studies are needed.

Denture stomatitis associated with small-colony variants of Staphylococcus aureus: a case report.

BACKGROUND: The small-colony variants (SCVs) of Staphylococcus aureus were isolated from persistent and recurrent infections, especially after placement of medical devices having direct contact with human tissues. The emergence of SCVs is a survival strategy of S. aureus which enables them to hide inside host's cells and induces a less severe immune response than to wild-type S. aureus. However, contrary to other medical devices, dental prosthesis as a surface potentially colonized by SCVs of S. aureus has not been examined thus far. We reported the first case of SCVs - S. aureus infection in denture wearer. CASE PRESENTATION: A 62-year-old woman with a complete removable acrylic denture presented extensive elevated erythematous lesions on the palate, compatible with denture stomatitis. The patient had a history of arterial hypertension, cigarette smoking and wearing denture at night. The fungal colonies, identified as Candida albicans, were cultured on Sabouraud agar. From three swabs (from hard palate mucosa, denture surface and angular cheilitis lesions) were cultured of pinpoint, clear, non-pigmented, and non-haemolytic colonies on Columbia agar. The small colonies turned out to be Gram-positive cocci, catalase-, Pastorex Staph Plus -, and clumping factor-positive, and oxidase-negative. Suspected phenotypically SCVs forms were definitively identified as S. aureus based on PCR amplification of species specific nuc and coa genes. Methicillin-resistance was verified by mecA gene detection. The isolates turned out to be susceptible to methicillin (MSSA) and resistant to gentamicin. The isolate was identified as menadione-auxotrophic variant. CONCLUSIONS: This case demonstrated that oral cavity in denture wearers may be a reservoir of small-colony variants of S. aureus, besides C. albicans. The prevalence of these bacteria and their role in the pathogenesis of oral diseases are not understood. Due to problems with their detection and identification, the true prevalence of oral SCVs may be underestimated.

Dissemination Of t437-SCCmecIV And Coagulase-Negative t037-SCCmecIII Types Among Borderline Oxacillin-Resistant Staphylococcus aureus Isolated From Skin Infections And Diabetic Foot Ulcers.

BACKGROUND: In a recent decade, the occurrence of S. aureus isolates with low-level oxacillin resistance, have been reported increasingly. The aim of this study was to estimate the prevalence of S. aureus with low-level of oxacillin resistance and to determine their molecular characteristics, including spa types, SCCmec types and presence of toxin genes. METHODS: A total of 249 S. aureus strains were analyzed. Antimicrobial susceptibility was preliminarily tested by the disk diffusion method, and further was verified with the E-test and agar dilution methods. All borderline oxacillin-resistant strains (BORSA) were screened for the mecA gene and virulence factors, including Panton-Valentine leukocidin (PVL). Staphylococcal cassette chromosome mec (SCCmec) typing and spa typing were also carried out. RESULTS: Twelve (4.8%) borderline oxacillin-resistant strains with MIC ≤4 µg/mL were identified. Almost all strains (11/12) were oxacillin-susceptible methicillin resistant S. aureus carrying mecA gene (OS-MRSA). Among the 12 bordeline strains, five spa types (t437, t037, t015, t216, t267) and two SCCmec types (III, IV) were identified, with the most prevalent being t437-SCCmecIV pvl-positive. The second most frequent spa type, t037-SCCmecIII, was sea-positive and did not produce coagulase. The majority of borderline strains originated from skin infections and diabetic foot ulcers and were multidrug-resistant (macrolides, lincosamides and chloramphenicol). CONCLUSION: This study demonstrated that S. aureus with borderline resistance to oxacillin represented primarily SCCmecIV spa type t437 and coagulase-negative SCCmecIII spa type t037 and were isolated from skin infections and diabetic foot ulcers.