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    Detection of Biofilm Production and Antimicrobial Susceptibility in Clinical Isolates of Acinetobacter baumannii and Pseudomonas aeruginosa
    (Институт за јавно здравје на Република Македонија = Institute of public health of Republic of Macedonia, 2022-08)
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    Todosovska Ristovska, Aneta
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    Lameski, Maja
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    Preshova, Ardian
    Acinetobacter baumannii and Pseudomonas aeruginosa are commensal which commonly colonize humans. As a result of their ubiquitous nature, reservoirs in hospital environment and resistance to many antimicrobial agents they are responsible for hospital – acquired infections. Additionally treatment of these infections is difficult because of the ability for biofilm formation. Aim of the paper was to determine the association between biofilm formation on medical devices and antibiotic resistance profile, compared to respiratory samples in clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa. Material and methods: The study comprised 50 clinical samples (36 from medical devices and 14 as а control group from respiratory secretions). Acinetobacter baumannii and Pseudomonas aeruginosa were identified by routine microbiological methods. Modification of the microtiter plate assay described by Stepanovic et al. was used to investigate the formation of biofilm. The antimicrobial susceptibility testing was performed according to EUCAST guidelines. Results: Of the 50 analyzed strains, 16 (32%) were non-biofilm producers, and 34 (68%) were producing biofilms. Out of these, 29 (58%) were from medical devices, and 5 (10%) from the control group. Acinetobacter baumannii showed biofilm formation in 19 (67.9%), of which 17 (60.7%) from medical devices, and 2 (7.1%) from control group. Pseudomonas aeruginosa produced biofilm in 15 (68.1%), of which 12 (54.5%) from medical devices, and 3 (13.6%) from the control group. Multidrug resistance was detected in 40 (80%). All strains of Acinetobacter baumannii were multidrug resistant (MDR). For Pseudomonas aeruginosa, 11 (73.3%) biofilm forming isolates were MDR, and 1 (14.2%) non-biofilm forming isolate was MDR. Conclusion: Biofilm production was higher in strains from medical devices. Eighty percent of isolates were MDR. This is a serious challenge for treatment of these hospital-acquired infections.
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    CHRONIC WOUNDS - MICROBIOLOGY AND BIOFILM FORMATION
    (Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, 2024)
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    Ahtarova, Biljana
    Introduction: Chronic wounds are wounds that do not heal for more than 4 weeks. Among the most important causes of wound chronicity are wound infection, biofilm formation with microbial agents that are resistant to antimicrobial agents. Aim: To investigate microbial agents in chronic wounds, their potential to form biofilm. Material and methods: This was a prospective study involving 24 patients from the University Clinic for in Skopje. Microbiological swabs were taken from patients with chronic wounds on the lower extremities,and were analyzed at the Institute of Microbiology and Parasitology, Faculty of Medicine in Skopje. The biofilm formation potential in isolates was determined using the tissue culture plate (TCP) biofilm detection method. Results: In 17 patients, at least one microorganism was isolated from 23 wounds, a total of 36, and in 7 patients there was no positive isolate from 7 wounds. Eighteen wounds (60%) had an isolate with the potential to form a biofilm. The following microorganisms from the wound samples were recorded: 17 (47%) isolates of the genus Staphylococcus, 4 (11%) isolates each from the genera Proteusand Pseudomonas, 3 (8%) isolates each from Enterococcusand Escherichia, 2 (6%) isolates from Streptococcus, 1 (3%) each isolate from the genera Acinetobacter, Enterobacterand Candida albicans. Conclusion: In 60% of the isolates, a biofilm formation with different degrees was confirmed. A statistically significant association was found between the degree of wound healing and isolates with low potential for biofilm formation, as well as the type of isolate.
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    BIOFILM IN CHRONIC DIABETIC FOOT ULCER-CASE REPORT
    (Macedonian Association of Anatomists and Morphologists, 2023)
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    Labacevska Gjatovska, Liljana
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    Dohceva Karajovanov, Ivana
    Diabetic foot ulcers(DFUs) as one of the most common complications in patients with diabetes mellitus are usually chronic wounds.The reason for its chronicity are infections and biofilm formation. We present a patient with diabetic foot neuropathic ulcer on the right foot. Microbiological swab showed isolates of bacteria and fungi, Candida albicans,Enterococcus and Acinetobacter which were tested for biofilm formation with microtiter plate assay. Biofilm mass was evaluated spectrophotometrically by measuring the absorbence of crystal violet. Enterococcus was with high potential of biofilm formation. Wound surface was measured every week for a period of one month and it was reduced for 23.93%. Ulcer was treated with peroral antibiotic and antifungal medications and standard wound care was performed. Microorganisms isolated from wound swabs showed mixed bacterial and fungal components. Current sudies show that relation in between this biofilm is still unclear. All of this is a key role in treating chronic wounds, making it a challenge for everyone not only in the field of making diagnosis , but also in the field of treatment.
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    MICROBIOME AND MYCOBIOME IN CHRONIC WOUNDS
    (Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, 2023)
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    Skin as the biggest organ with protective function in the human body, makes an equilibrium between microbial communities and immune system. Skin microbiome is defined as the genome of microorganisms found on the skin with which microorganisms have a complex relationship. Microbiota of healthy skin consists of resident and transient microorganisms. Two most common factors for delayed healing process in chronic wounds are infection and biofilm formation. Thus, it is important to analyze microbiome and mycobiome of chronic wounds.
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    Susceptibility of strong biofilm-producing uroisolates in planktonic state vs. Biofilm growth mode.
    (Macedonian Association of Anatomists and Morphologists (MAAM), 2021)
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    Jankoska, G
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    Bojan Labachevski
    Introduction: Biofilms are defined as functional consortiums of microorganisms, attached to a surface, enclosed in self-produced hydrated polymeric matrix. Between microorganisms in biofilm and free floating microorganisms of the same species there are essential differences regarding: specific gene expression, ability for intercellular communication via biochemical signaling molecules, antimicrobial resistance. These biofilm features play a key role in the development of chronic and antibiotics tolerant infections. The ability of uropathogens to cause relapses of UTI directly correlates with their genetic capability for biofilm production on catheter surfaces or uroepithelium and with the innate recalcitrance to treatment of the biofilm itself. Objective: To investigate the differences in pathogens resistance in relation to biofilm expression phenotype by comparing the antibiotic susceptibility of sessile cells and their planktonic counterpart, for biofilm forming bacteria isolated from patients with UTI. Materials and methods: The study included 120 urine samples from outpatients, from both genders, over 18 years of age, with suspected UTI, referred for microbiological examination of urine at the Institute of microbiology and parasitology, Medical faculty, Skopje. A total number of 80 bacterial species, isolated in monobacterial culture were examined for biofilm production. For biofilm cultivation and biofilm biomass determination, adherence assay on 96-well microtitre plate and semi-quantitative spectrophotometric method were used. For 25 selected strong biofilm producers, antibiotic susceptibility to ciprofloxacin and sulfametoxasole trimetoprime of planktonic cells was tested by microdilution assay and compared with the antimicrobial sensitivity of bacterial biofilms (performed with the Calgary Biofilm Device) Results: Distribution of susceptibilities, regarding the frequency of the sensitive, intermediate and resistant bacteria showed statistically significant difference in the antibiotic susceptibility of planktonic cells vs. their sessile counterparts (p<0.01) / Fisher ́s Exact p. Results showed that bacteria are more resistant in biofilm communities as compared to planktonic form by comparison of MIC & MBEC assay. Conclusion: Attached bacterial communities (biofilms) present with an innate deficit of antibiotic susceptibility not registered in the same bacteria grown at planktonic state. Minimal biofilm eradication concentrations, determined applying the Calgary Biofilm Device, reveal that treatment of biofilms requires 20 to 1,200 times concentrations of a certain antibiotic to achieve the desired antimicrobial effect compared with planktonic cells of the same organisms which urges future changes in the conventional therapeutical approach.