Browsing by Author "Vuković, Dragana (7005414538)"

The tube test and the microtiter-plate test are the most frequently used techniques for quantifying biofilm formation, an important indicator for the pathogenicity of staphylococci. The purpose of the present study was to develop a modified microtiter-plate technique for quantification of biofilm formation. This technique involves fixing the bacterial film with methanol, staining with crystal violet, releasing the bound dye with 33% glacial acetic acid, and measuring the optical density (OD) of the solution at 570 nm by using an enzyme immunosorbent assay reader. Biofilm formation of 30 Staphylococcus strains was estimated by the tube test, the standard microtiter-plate test and the modified microtiter-plate test. The modified microtiter-plate test, as a quantitative assay, is superior to the tube test in terms of objectivity and accuracy. It is also superior to the standard microtiter-plate test because it enables indirect measuring of bacteria attached both to the bottom and to the walls of the wells, while in the standard test only the dye bound to the bacteria adhered to the bottom of the wells is spectrophotometrically registered. Highly significant differences between OD values obtained by the standard microtiter-plate test and those obtained by the modified test suggest that large number of bacteria were attached to the walls of the wells. Therefore, the modification of the standard microtiter-plate test by introduction of an additional step of decolorization by acetic acid seems to be a useful improvement of the technique. © 2000 Elsevier Science B.V.

The tube test and the microtiter-plate test are the most frequently used techniques for quantifying biofilm formation, an important indicator for the pathogenicity of staphylococci. The purpose of the present study was to develop a modified microtiter-plate technique for quantification of biofilm formation. This technique involves fixing the bacterial film with methanol, staining with crystal violet, releasing the bound dye with 33% glacial acetic acid, and measuring the optical density (OD) of the solution at 570 nm by using an enzyme immunosorbent assay reader. Biofilm formation of 30 Staphylococcus strains was estimated by the tube test, the standard microtiter-plate test and the modified microtiter-plate test. The modified microtiter-plate test, as a quantitative assay, is superior to the tube test in terms of objectivity and accuracy. It is also superior to the standard microtiter-plate test because it enables indirect measuring of bacteria attached both to the bottom and to the walls of the wells, while in the standard test only the dye bound to the bacteria adhered to the bottom of the wells is spectrophotometrically registered. Highly significant differences between OD values obtained by the standard microtiter-plate test and those obtained by the modified test suggest that large number of bacteria were attached to the walls of the wells. Therefore, the modification of the standard microtiter-plate test by introduction of an additional step of decolorization by acetic acid seems to be a useful improvement of the technique. © 2000 Elsevier Science B.V.

This study investigated the prevalence of aminoglycoside resistance and genes encoding aminoglycoside-modifying enzymes in members of the Staphylococcus sciuri group. A total of 304 S. sciuri group member isolates (284 S. sciuri, 12 S. lentus, and 8 S. vitulinus) from humans (n = 34), animals (n = 133), and environmental sources (n = 137; out-hospital and hospital environment, food) were examined for their susceptibility to amikacin, gentamicin, isepamicin, kanamycin, neomycin, netilmicin, sisomicin, streptomycin, and tobramycin. The overall prevalence of resistance to aminoglycosides was low at 12.1%. Resistance to single aminoglycosides ranged from 0% to 7.2%. The aac(6′)-Ie/ aph(2″), ant(4′)-Ia, and aph(3′)-IIIa genes, either alone or in combination, were found in 16 out of 19 isolates showing resistance to nonstreptomycin aminoglycosides. Among the 22 isolates that showed resistance to streptomycin, the genes str and ant(6)-Ia were identified in 18 and 4 isolates, respectively. © Mary Ann Liebert, Inc.

This study investigated the prevalence of aminoglycoside resistance and genes encoding aminoglycoside-modifying enzymes in members of the Staphylococcus sciuri group. A total of 304 S. sciuri group member isolates (284 S. sciuri, 12 S. lentus, and 8 S. vitulinus) from humans (n = 34), animals (n = 133), and environmental sources (n = 137; out-hospital and hospital environment, food) were examined for their susceptibility to amikacin, gentamicin, isepamicin, kanamycin, neomycin, netilmicin, sisomicin, streptomycin, and tobramycin. The overall prevalence of resistance to aminoglycosides was low at 12.1%. Resistance to single aminoglycosides ranged from 0% to 7.2%. The aac(6′)-Ie/ aph(2″), ant(4′)-Ia, and aph(3′)-IIIa genes, either alone or in combination, were found in 16 out of 19 isolates showing resistance to nonstreptomycin aminoglycosides. Among the 22 isolates that showed resistance to streptomycin, the genes str and ant(6)-Ia were identified in 18 and 4 isolates, respectively. © Mary Ann Liebert, Inc.

Objective: To characterize the antitoxoplasma activity of clindamycin in a murine model of acute toxoplasmosis. Methods: Rates of survival and mean survival times of Swiss Webster mice infected intraperitoneally with 106-102 tachyzoites of the RH strain of Toxoplasma gondii treated with clindamycin or sulfamethoxazole (positive control) or untreated (negative control) were compared. Survivors were submitted to examination of untreated brain tissue preparations, intraperitoneal and peroral subinoculations of brain tissue homogenates into fresh mice, and to pathohistology, including immunohistochemistry, of brain and lungs. Results: The effect of clindamycin treatment (400 mg/kg/day) on infected Swiss Webster mice was inoculum size dependent, ranging from no survivals in animals infected with 106 parasites, to 100% survivals with an inoculum of 102. Treatment initiated 24 h before and at time of infection prolonged mean survival times comparably to sulfamethoxazole, and significantly when compared to untreated controls. In contrast, treatment initiated 48 h postinfection with an inoculum of 106 did not postpone death. In the clindamycin-treated survivors, there was no biological or histologic evidence for the persistence of toxoplasma. Conclusions: The results obtained show that at an appropriate parasite dose/drug dose ratio, clindamycin is strongly toxoplasmacidal in a murine model of acute toxoplasmosis.

[No abstract available]

A total of 28 staphylococcal isolates from human clinical specimens belonging to the Staphylococcus sciuri group were identified and characterized. The API Staph and ID32 STAPH correctly identified S. sciuri and S. lentus but not S. vitulinus strains. Identification to the subspecies level was possible only by a PCR-based method.

In this study, the focus was on the effects of sub-MICs of the antibiotics on adherence, hydrophobicity, and biofilm formation by two groups of Streptococcus pyogenes strains, which were responsible for different clinical cases. The aim of this study was to explore the effects of sub-MICs of penicillin, ceftriaxone, erythromycin, and clindamycin on adherence, surface hydrophobicity, and biofilm biomass in two selected collections of group A streptococcus (GAS): strains isolated from carriers (CA) and strains isolated from patients with tonsillopharyngitis (TPh). Isolates were tested for hydrophobicity to xylene, adherence, and biofilm production in uncoated microtiter plates before and after treatment with 1/2 and 1/4 MICs of antibiotics. Penicillin reduced adherence and biofilm production in TPh strains, whereas ceftriaxone diminished adherence and biofilm formation in CA group. On the contrary, clindamycin enhanced adherence and biofilm production in both groups of strains. Erythromycin did not significantly alter adherence, but triggered biofilm production in both groups of isolates. Hydrophobicity of both groups of strains was significantly reduced after exposure to all antibiotics. Beta-lactams displayed anti-biofilm activity; penicillin diminished both adherence and biofilm production in TPh strains, whereas ceftriaxone reduced it in strains isolated from CA. © 2018 Akadémiai Kiadó, Budapest.

In this study, the focus was on the effects of sub-MICs of the antibiotics on adherence, hydrophobicity, and biofilm formation by two groups of Streptococcus pyogenes strains, which were responsible for different clinical cases. The aim of this study was to explore the effects of sub-MICs of penicillin, ceftriaxone, erythromycin, and clindamycin on adherence, surface hydrophobicity, and biofilm biomass in two selected collections of group A streptococcus (GAS): strains isolated from carriers (CA) and strains isolated from patients with tonsillopharyngitis (TPh). Isolates were tested for hydrophobicity to xylene, adherence, and biofilm production in uncoated microtiter plates before and after treatment with 1/2 and 1/4 MICs of antibiotics. Penicillin reduced adherence and biofilm production in TPh strains, whereas ceftriaxone diminished adherence and biofilm formation in CA group. On the contrary, clindamycin enhanced adherence and biofilm production in both groups of strains. Erythromycin did not significantly alter adherence, but triggered biofilm production in both groups of isolates. Hydrophobicity of both groups of strains was significantly reduced after exposure to all antibiotics. Beta-lactams displayed anti-biofilm activity; penicillin diminished both adherence and biofilm production in TPh strains, whereas ceftriaxone reduced it in strains isolated from CA. © 2018 Akadémiai Kiadó, Budapest.

Staphylococcus sciuri is a principally animal-associated bacterial species, but its clinical relevance for humans is increasing. Our study aimed to provide the first insight into the prevalence of this bacterium in a hospital environment. A 3-month surveillance was conducted in a hospital located in Belgrade, Serbia, and 1,028 samples taken from hands of medical personnel, medical devices, and various hospital surfaces were screened for S. sciuri presence. In total, 108 isolates were obtained, which resulted in a relatively high rate of colonization (10.5%). These isolates, along with 7 S. sciuri strains previously isolated in the same hospital (n = 115), were phenotypically and genotypically characterized. Antimicrobial susceptibility testing revealed that 73% of the strains were resistant to one or more antibiotics, with 4.3% strains displaying multiresistance. Examination of 16S-23S ribosomal DNA intergenic spacer length polymorphism identified the strains at the subspecies level, and 74 (64.3%) strains of S. sciuri subsp. sciuri, 37 (32.2%) strains of S. sciuri subsp. rodentium, and 4 (3.5%) strains of S. sciuri subsp. carnaticus were established. Pulsed-field gel electrophoresis (PFGE) analysis showed 21 distinct pulsotypes, including 17 main types and 4 subtypes. One dominant cluster with 62 strains was found, while 19 (90.5%) of the PFGE types and subtypes identified had 5 or fewer strains. The predominance of small PFGE clusters suggests that the ubiquitous presence of S. sciuri in the outside environment presents the continuous source for colonization of the hospital environment. The presence of one dominant PFGE cluster of strains indicates that some S. sciuri strains may be capable for adaptation to hospital environment conditions and continuous existence in this environment. Copyright © 2005, American Society for Microbiology. All Rights Reserved.

During a 3-year study period, 32,741 urine samples were analyzed for the presence of members of the Staphylococcus sciuri group (S. sciuri, S. lentus, and S. vitulinus), and 13 isolates were identified. They presented 0.79% of the total number of coagulase-negative staphylococci isolated. One case of symptomatic urinary tract infection and five possible cases of asymptomatic bacteriuria caused by these bacteria were established. It is noteworthy, however, that over 50% of the isolates originated from hospitalized patients.

Lyme endocarditis is extremely rare manifestation of Lyme disease. The clinical manifestations of Lyme endocarditis are non-specific and can be very challenging diagnosis to make when it is the only manifestation of the disease. Until now, only a few cases where reported. Physicians should keep in mind the possibility of borrelial etiology of endocarditis in endemic areas. Appropriate valve tissue sample should be sent for histopathology, culture, and PCR especially in case of endocarditis of unknown origin PCR on heart valve samples is recommended. With more frequent PCR, Borrelia spp. may be increasingly found as a cause of infective endocarditis. Prompt diagnosis and treatment of Lyme carditis may prevent surgical treatment and pacemaker implantations. Due to climate change and global warming Lyme disease is a growing problem. Rising number of Lyme disease cases we can expect and rising number of Lyme endocarditis. © Copyright © 2020 Nikolić, Boljević, Bojić, Veljković, Vuković, Paglietti, Micić and Rubino.

Lyme endocarditis is extremely rare manifestation of Lyme disease. The clinical manifestations of Lyme endocarditis are non-specific and can be very challenging diagnosis to make when it is the only manifestation of the disease. Until now, only a few cases where reported. Physicians should keep in mind the possibility of borrelial etiology of endocarditis in endemic areas. Appropriate valve tissue sample should be sent for histopathology, culture, and PCR especially in case of endocarditis of unknown origin PCR on heart valve samples is recommended. With more frequent PCR, Borrelia spp. may be increasingly found as a cause of infective endocarditis. Prompt diagnosis and treatment of Lyme carditis may prevent surgical treatment and pacemaker implantations. Due to climate change and global warming Lyme disease is a growing problem. Rising number of Lyme disease cases we can expect and rising number of Lyme endocarditis. © Copyright © 2020 Nikolić, Boljević, Bojić, Veljković, Vuković, Paglietti, Micić and Rubino.

Introduction Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) present the growing problem in the whole world. Carriage of MRSA is most frequent in the nose, and medical students come in contact both with patients and different persons in the community. Therefore, they may be significant for the transmission of MRSA from hospitals to outof- hospital communities and vice versa. Objective The aim of this study was to establish the carriage rate among students of the second, third and fourth year of study at the School of Medicine in Belgrade and to analyze their genotypic and phenotypic characteristics. Methods In total 533 nasal samples were taken. The samples were incubated in Trypcase-soy broth supplemented with 6.5% NaCl, and thereafter the swabs were inoculated on mannitol salt agar supplemented with 2 μg/mL of oxacillin. The presence of nuc, mecA and Panton-Valentine leukocidin genes was examined by PCR. The characteristics of the MRSA strains were determined using: antibiotic susceptibility testing by Vitek2 System, SCCmec, agr typing and MLST. Results MRSA was isolated from two of 533 investigated samples (0.37%). MRSA were isolated from the students of the second and third year of study. Profiles of strains were: ST80 (SCCmec type IV, agr type 3) and ST152 (SCCmec type V, agr type 1). MRSA strains were multiresistant. Conclusion The nasal carriage rate of MRSA in population of medical students of the first year of study in Belgrade is low. Genotypic and phenotypic characteristics of MRSA strains indicate their community origin. MLST typing revealed that isolates belong to ST80 and ST152.

[No abstract available]

The scope of the antibacterial effects of plasma-activated water (PAW) is not yet fully comprehended. We investigated the activity of PAW produced by the in-house 3-pin atmospheric pressure plasma jet against carbapenem-resistant Klebsiella pneumoniae and vancomycin-resistant Enterococcus faecalis, with a focus on PAW’s potential to promote susceptibility to conventional antibiotics in these bacteria. Bacterial inactivation was determined by the colony count after 15 and 60 min PAW treatments. Minimum inhibitory concentrations (MICs) measured following repeated exposures to PAW across multiple generations of bacteria enabled the assessment of changes in susceptibility to antibiotics. The PAW’s efficacy was also analyzed through the detection of intracellular reactive oxygen and nitrogen species in treated bacteria. Time-dependent significant inactivation efficiency against K. pneumoniae was observed (log reduction 6.92 ± 0.24 after 60 min exposure), while effects on E. faecalis were limited. PAW demonstrated potential to decrease the MICs of crucial antibiotics. Namely, a 50 to 62.5% decrease in the MICs of colistin against K. pneumoniae and a 25% reduction in the MICs of vancomycin against enterococci were recorded. We found a significant increase in the superoxide anion concentration in K. pneumoniae and E. faecalis cells after PAW treatments. This study indicates that PAW’s inactivating efficacy coupled with the capacity for the potentiation of antibiotic effects is a promising combination against multidrug-resistant bacteria. © 2025 by the authors.

The details of all steps involved in the quantification of biofilm formation in microtiter plates are described. The presented protocol incorporates information on assessment of biofilm production by staphylococci, gained both by direct experience as well as by analysis of methods for assaying biofilm production. The obtained results should simplify quantification of biofilm formation in microtiter plates, and make it more reliable and comparable among different laboratories. Copyright © Apmis 2007.

This study aimed to characterize the resistance profiles of the Staphylococcus sciuri group members to macrolides, lincosamides, streptogramins (MLS antibiotics), and linezolid upon analysis of large series of isolates that included 162 S. sciuri isolates, nine S. lentus, and one S. vitulinus. The evaluation of their susceptibility by disk diffusion and agar dilution methods, along with PCR detection of the resistance genes erm(A), erm(R), erm(C), mef(A), lnu(A), and lnu(B), were performed. Resistance to macrolides was detected in 10 (5.8%) tested strains, with three and six isolates exhibiting constitutive and inducible MLSB resistance phenotypes, respectively. Resistance mediated by active efflux was detected in one strain. The presence of genes conferring resistance, namely erm(B) or erm(C), was detected in two strains. All tested strains were susceptible to pristinamycin and linezolid. Of 172 tested strains, 70.9% were resistant and 26.2% had intermediary resistance to lincomycin, whereas 1.7% were resistant and 50% had intermediary resistance to clindamycin. The lnu(A) gene was detected in two strains only. The great majority of the tested S. sciuri strains (153 out of 162; 94.4%) presumably exhibited LSA phenotype because they did not carry lnu genes nor displayed constitutive MLSB resistance, but still showed intermediate resistance or resistance to lincomycin (MICs of 4, 8, 16, and 32 μg/ml). The results obtained indicate that S. sciuri may be naturally resistant to lincomycin. Expression of a novel type of inducible resistance to lincosamides, induced by erythromycin in erythromycin-susceptible strains, was observed in the S. sciuri group isolates. © Mary Ann Liebert, Inc.

This study aimed to characterize the resistance profiles of the Staphylococcus sciuri group members to macrolides, lincosamides, streptogramins (MLS antibiotics), and linezolid upon analysis of large series of isolates that included 162 S. sciuri isolates, nine S. lentus, and one S. vitulinus. The evaluation of their susceptibility by disk diffusion and agar dilution methods, along with PCR detection of the resistance genes erm(A), erm(R), erm(C), mef(A), lnu(A), and lnu(B), were performed. Resistance to macrolides was detected in 10 (5.8%) tested strains, with three and six isolates exhibiting constitutive and inducible MLSB resistance phenotypes, respectively. Resistance mediated by active efflux was detected in one strain. The presence of genes conferring resistance, namely erm(B) or erm(C), was detected in two strains. All tested strains were susceptible to pristinamycin and linezolid. Of 172 tested strains, 70.9% were resistant and 26.2% had intermediary resistance to lincomycin, whereas 1.7% were resistant and 50% had intermediary resistance to clindamycin. The lnu(A) gene was detected in two strains only. The great majority of the tested S. sciuri strains (153 out of 162; 94.4%) presumably exhibited LSA phenotype because they did not carry lnu genes nor displayed constitutive MLSB resistance, but still showed intermediate resistance or resistance to lincomycin (MICs of 4, 8, 16, and 32 μg/ml). The results obtained indicate that S. sciuri may be naturally resistant to lincomycin. Expression of a novel type of inducible resistance to lincosamides, induced by erythromycin in erythromycin-susceptible strains, was observed in the S. sciuri group isolates. © Mary Ann Liebert, Inc.

Objectives: To determine the incidence of toxoplasmic encephalitis (TE) in patients with acquired immunodeficiency syndrome (AIDS) in Yugoslavia and to analyze the contribution of sex, age, human immunodeficiency virus (HIV) transmission risk factors, CD4+ T-cell count, and systemic Pneumocystis carinii pneumonia (PCP) prophylaxis as risk factors for the development of TE. Design: Retrospective chart review of 288 (72%) of the 399 AIDS patients diagnosed in Yugoslavia during the 5-year period 1991 to 1995. Inclusion criteria included serology for toxoplasmosis performed prior to the TE episode. Results: Toxoplasma gondii-specific IgG antibodies were detected in 127 of the 288 AIDS patients (44.1%) tested serologically (including all with neurologic abnormalities). A total of 31 patients developed TE, indicating a TE overall attack rate of 7.8%. Of these, 29 TE episodes occurred in the T. gondii-seropositive patients (22.8%), significantly more often than in the T. gondii-seronegative (χ2 = 34.423, P < 0.005), among which TE occurred in two (2/161, 1.24%). By survival analysis, the cumulative incidence of TE in the T. gondii-seropositive patients was 32.7% for 60 months (30% for 24 months). Although not associated with age, sex, or HIV transmission risk factor, the risk for TE increased with the decrease in the CD4+ T-cell count (relative risk [RR] = 0.276, 95% confidence interval [Cl] = 0.198-0.673, P = 0.0013), and was reduced in patients on systemic PCP prophylaxis (RR = 0.222, 95% Cl = 0.068-0.642, P = 0.006). Conclusions: In areas of high exposure to T. gondii, such asYugoslavia, timely serology for toxoplasmosis in patients with AIDS and subsequent drug prophylaxis to prevent reactivation in seropositive patients is recommended.