Effect of Camel Peptide on the Biofilm of Staphylococcus epidermidis and Staphylococcus haemolyticus Formed on Orthopedic Implants.

The increasing bacterial drug resistance and the associated challenges in the treatment of infections warrant the search for alternative therapeutic methods. Hope is placed in antimicrobial peptides, which have a broad spectrum of action and are effective against strains which are resistant to conventional antibiotics. Antimicrobial peptides are also tested for their efficacy in the treatment of infections associated with the formation of biofilm. The aim of the present study was to examine the effect of Camel peptide on S. epidermidis and S. haemolyticus adhesion to and formation of biofilm on steel cortical bone screws and also on the process of reducing mature biofilm in orthopedic implants. The tests were performed on steel implants for osteosynthesis. The MIC value and MBEC values of the peptide were determined using the microdilution method in microtiter plates. The effect of the peptide on adhesion and biofilm formation, as well as on the activity on the preformed biofilm, was evaluated using quantitative methods and confocal microscopy. The presented research results indicate that the peptide exhibits very good antimicrobial properties against the analyzed strains. Concentrations above MIC reduced biofilm in the range of 90-99%.

Preservation fluids of heritage anatomical specimens - a challenge for modern science. Studies of the origin, composition and microbiological contamination of old museum collections.

Anatomical museums preserve specimens of great historical value and undiscovered scientific potential. However, frequently these collections lack documentation of the techniques of preparation and the composition of preservative substances (conservation principles). This poses a huge problem for the care and preservation of these materials, more so because understanding this issue requires knowledge of fundamentals from different scientific disciplines. The aim of the research was to obtain information about the composition of substances used to preserve historic specimens, as well as to conduct a microbiological assessment of the specimens to detect possible factors causing their deterioration. Furthermore, we wanted to fill an existing gap in the literature, as there is a lack of reports on analytical methods that could be successfully applied by anatomists involved in the daily care of museum collections in human anatomy departments. The starting point was the analysis of the sources and history of the collections, on which basis the choice of research methods was made. Methods based on simple chemical reactions and specialised methods (such as gas chromatography-tandem mass spectrometry, Fourier transform infrared spectroscopy, inductively coupled plasma optical emission spectroscopy) were used in the analyses of the composition of fluids. Microbiological analyses were based on culture and isolation methods, analysis of microscopy slides and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry analysis. As a result of these analyses, some components of the preservative mixtures and their concentrations were determined. The presence of methanol, ethanol, formaldehyde and glycerol was detected, among other chemicals. The concentrations of these substances were different between the samples and their determination required the use of a variety of methods suitable for the individual components of the preservative mixture. In microbiological tests, both bacteria and fungi were isolated from swabs taken from anatomical specimens. The bacterial flora was less numerous than the fungal flora. Among the bacteria, environmental Gram-positive Bacillus cereus, Bacillus thuringiensis and a rare bacterium of the Cupriavidus genus were isolated, whereas among the fungal organisms, the yeast-like fungi Candida boidinii and Geotrichum silvicola as well as mould fungi Penicillium sp. and Fusarium sp. were detected. However, the microscopic evaluation showed a greater diversity of microorganisms, which may be related to the fact that many environmental bacteria cannot be cultured using classical methods, but can be observed under the microscope. The results of the research made it possible to draw conclusions about the mutual influence of physical, chemical, and microbiological factors on the condition of historical anatomical specimens. In the course of the research, information was obtained on the processes which could have taken place during the storage of these collections. Maintaining the integrity of a container housing a preserved anatomical specimen has a major impact on maintaining the concentration of preservative fluid and keeping the specimen environment sterile. Many conservation procedures for historical specimens carried out nowadays pose a risk of destroying valuable specimens, as well as a health risk for the person carrying out the work. The exploration of the topic of conservation of anatomical specimens, especially those that lack documentation of their origin, is a key issue in current research on historical collections of anatomical specimens.

Current State of Knowledge about Role of Pets in Zoonotic Transmission of SARS-CoV-2.

Pets play a crucial role in the development of human feelings, social life, and care. However, in the era of the prevailing global pandemic of COVID-19 disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), many questions addressing the routes of the virus spread and transmission to humans are dramatically emerging. Although cases of SARS-CoV-2 infection have been found in pets including dogs, cats, and ferrets, to date there is no strong evidence for pet-to-human transmission or sustained pet-to-pet transmission of SARS-CoV-2. However, an increasing number of studies reporting detection of SARS-CoV-2 in farmed minks raises suspicion of potential viral transmission from these animals to humans. Furthermore, due to the high susceptibility of cats, ferrets, minks and hamsters to COVID-19 infection under natural and/or experimental conditions, these animals have been extensively explored as animal models to study the SARS-CoV-2 pathogenesis and transmission. In this review, we present the latest reports focusing on SARS-CoV-2 detection, isolation, and characterization in pets. Moreover, based on the current literature, we document studies aiming to broaden the knowledge about pathogenicity and transmissibility of SARS-CoV-2, and the development of viral therapeutics, drugs and vaccines. Lastly, considering the high rate of SARS-CoV-2 evolution and replication, we also suggest routes of protection against the virus.

Identification of Fungi Isolated from Oral Cavity of Patients with HIV Using MALDI-TOF MS.

BACKGROUND: A growing incidence of invasive fungal infections, especially among immunocompromised patients, has given increased significance to microbiological diagnostics of yeast-like fungi. More accurate and faster fungi identification methods that can compete with classical methods are being searched for. In this paper, classical microbiological methods are compared to MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). METHODS: The diagnostic material was collected from buccal mucosa from 98 adults, including 69 with HIV. Only positive cultures were included in the study. RESULTS: Matching results were obtained in 45 samples, and there were nonmatching results in 35 samples, with the majority of these in the study group, constituting 50% of identifications within this group. A particularly common mistake resulting from the use of classical methods is the false identification of C. dubliniensis as C. albicans. Additionally, C. tropicalis proves to be difficult to identify. CONCLUSIONS: Our results and literature data suggest that MALDI-TOF MS should be considered an effective alternative to classical methods in terms of fungi identification, especially among HIV-positive patients, due to the different morphology of fungal colonies.

Oral cavity fungal flora among HIV-positive people.

BACKGROUND: Immunosuppressed patients, also those who are HIV-positive patients, are susceptible to oral cavity fungal infections. AIM OF STUDY: In this study, we aimed to show differences in qualitative composition of oral cavity flora between HIV-positive people and healthy controls and identify factors which affect fungal oral cavity flora. MATERIALS AND METHODS: The study group contained HIV-positive people and a control group of healthy people. All cultured species were analysed using MALDI-TOF MS. RESULTS: More HIV-positive people had two or more fungus species present than controls (p=0.008). Seven species were cultured in the study group compared to three in the control group. Smoking was associated with higher prevalence of C. albicans (p=0.03), C. glabrata (p=0.026), C. tropicalis (p=0.01). Dental prosthesis or braces was also associated with presence of more species (p=0.04).The lower level of lymphocytes CD4+ was not associated with fungus presence in oral cavity. CONCLUSIONS: HIV infection is associated with changes to oral cavity fungal flora. Given the higher number of non-albicans species among HIV-positive patients it is important to individually choose a treatment for such patients' fungal infections. Proper oral hygene and not smoking can reduce prevalence of fungi in oral cavity. Patients' immunological status did not have an impact on the frequency of Candida species isolation from the oral cavity.

Methods Used for the Eradication of Staphylococcal Biofilms.

Staphylococcus aureus is considered one of the leading pathogens responsible for community and healthcare-associated infections. Among them, infections caused by methicillin-resistant strains (MRSA) are connected with ineffective or prolonged treatment. The therapy of staphylococcal infections faces many difficulties, not only because of the bacteria's resistance to antibiotics and the multiplicity of virulence factors it produces, but also due to its ability to form a biofilm. The present review focuses on several approaches used for the assessment of staphylococcal biofilm eradication. The methods described here are successfully applied in research on the prevention of biofilm-associated infections, as well as in their management. They include not only the evaluation of the antimicrobial activity of novel compounds, but also the methods for biomaterial functionalization. Moreover, the advantages and limitations of different dyes and techniques used for biofilm characterization are discussed. Therefore, this review may be helpful for those scientists who work on the development of new antistaphylococcal compounds.

Use of the Real Time xCelligence System for Purposes of Medical Microbiology.

Roche's xCelligence impedance-measuring instrument is one of a few commercially available systems of such type. According to the best knowledge of authors, instrument was tested so far only for eukaryotic cell research. The aim of this work was to estimate xCELLigence suitability for the microbiological tests, including (i) measurement of morphological changes in eukaryotic cells as a result of bacterial toxin activity, (ii) measurement of bacterial biofilm formation and (iii) impact of antiseptics on the biofilm structure. To test the infuence of bacterial LT enterotoxin on eukaryotic cell lines, Chinese Hamster Ovary (CHO) cell line and reference strain Escherichia coli ATTC 35401 were used. To investigate Roche's instrument ability to measure biofilm formation and impact of antiseptics on its development, Staphylococcus aureus ATTC6538 reference strain was used. The data generated during the experiments indicate excellent ability of xCelligence instrument to detect cytopathic effect caused by bacterial LT endotoxin and to detect staphylococcal biofilm formation. However, interpretation of the results obtained during real-time measurement of antiseptic's bactericidal activity against staphylococcal biofilm, caused many difficulties. xCelligence instrument can be used for real-time monitoring of morphological changes in CHO cells treated with bacterial LT enterotoxin and for real-time measurement of staphylococcal biofilm formation in vitro. Further investigation is necessary to confirm suitability of system to analyze antiseptic's antimicrobial activity against biofilm in vitro.