Successive Acanthamoeba Corneal Isolates Identified in Poland Monitored in Terms of In Vitro Dynamics.

BACKGROUND: Amoebae of the genus Acanthamoeba cause a sight-threatening infection called Acanthamoeba keratitis. It is considered a rare disease in humans but poses an increasing threat to public health worldwide, including in Poland. We present successive isolates from serious keratitis preliminary examined in terms of the identification and monitoring of, among others, the in vitro dynamics of the detected strains. METHODS: Clinical and combined laboratory methods were applied; causative agents of the keratitis were identified at the cellular and molecular levels; isolates were cultivated in an axenic liquid medium and regularly monitored. RESULTS: In a phase-contrast microscope, Acanthamoeba sp. cysts and live trophozoites from corneal samples and in vitro cultures were assessed on the cellular level. Some isolates that were tested at the molecular level were found to correspond to A. mauritanensis, A. culbertsoni, A. castellanii, genotype T4. There was variability in the amoebic strain dynamics; high viability was expressed as trofozoites' long duration ability to intense multiply. CONCLUSIONS: Some strains from keratitis under diagnosis verification and dynamics assessment showed enough adaptive capability to grow in an axenic medium, allowing them to exhibit significant thermal tolerance. In vitro monitoring that was suitable for verifying in vivo examinations, in particular, was useful to detect the strong viability and pathogenic potential of successive Acanthamoeba strains with a long duration of high dynamics.

Influence of Exposure Parameters and Implant Position in Peri-Implant Bone Assessment in CBCT Images: An In Vitro Study.

The aim of this study was to assess the impact of dimensional distortion and its changes with modification of exposure setting parameters on the measurements of peri-implant bone margin. Ten titanium dental implants (InKone Primo, Global D, Paris, France) were placed in two prepared bovine ribs. Two bone models and an implant-with-transfer model were scanned with 3shape E4 (3shape, Copenhagen, Denmark) laboratory scanner. Cone beam computed tomography (CBCT) images of two bone models were taken with different values of voltage (60, 70, 80, 90 kV), tube current (4, 10 mA) and voxel size (200, 300 µm). All the data were superimposed using planning software, and the measurements of buccal bone thickness in two selected regions were performed both using CBCT and scan cross-sections. The mean squared error (MSE) being the squared differences between measurements was used in the accuracy assessment of the CBCT device. A one-way ANOVA revealed significant differences between voltage and MSE (p = 0.044), as well as implant position and MSE (p = 0.005). The distortions of measurements depend on bone margin thickness, and the higher the distance to measure, the higher the error. Accurate measurements of buccal bone thickness (MSE below 0.25) were achieved with voltage values of 70, 80, and 90 kV.

The Impact of Cone-Beam Computed Tomography Exposure Parameters on Peri-Implant Artifacts: A Literature Review.

Cone-beam computed tomography (CBCT) amounts to an excellent diagnostic tool to evaluate the peri-implant bone thickness in all dimensions. Despite the unquestionable advantages of CBCT, potential artifacts around dental implants might disturb the proper assessment of the surrounding structures. The artifacts may mask osseointegration, shallow bone defects, and other types of radiolucency, which make it difficult to establish an early diagnosis of bone loss. Proper diagnosis of bone defect is necessary to decide about surgical intervention. The aim of this literature review is to assess the CBCT exposure causing artifacts on the peri-implant structures. An electronic search of MEDLINE (PubMed) database includes studies published before July 2021 and supplemented by manual research. Clinical, ex vivo, in vitro, and animal studies evaluating the relationship between exposition parameters and occurrence of artifacts around the dental implant in CBCT studies were included. A literature review revealed that kilovoltage, tube current, and field of view may affect the occurrence of artifacts around dental implants, all of which would compromise radiological evaluation. Therefore, it is feasible to reduce the incidence of artifacts and improve the image quality by appropriate modification of the exposure parameters. However, the reduction of artifacts is often associated with a significant increase in radiation exposure; hence, an effort should be made to minimize the radiation dose in line with the ALARA (as low as reasonably achievable) principle.

Examination of Oral Microbiota Diversity in Adults and Older Adults as an Approach to Prevent Spread of Risk Factors for Human Infections.

The oral cavity environment may be colonized by polymicrobial communities with complex, poorly known interrelations. The aim of this study was to determine oral microbiota diversity in order to prevent the spread of infectious microorganisms that are risk factors for human health complications in patients requiring treatment due to various disabilities. The study examined Polish adults aged between 40 and 70 years; parasitological, microbiological, and mycological data collected before treatment were analyzed. The diversity of oral microbiota, including relatively high prevalences of some opportunistic, potentially pathogenic strains of bacteria, protozoans, and fungi detected in the patients analyzed, may result in increasing risk of disseminated infections from the oral cavity to neighboring structures and other organs. Increasing ageing of human populations is noted in recent decades in many countries, including Poland. The growing number of older adults with different oral health disabilities, who are more prone to development of oral and systemic pathology, is an increasing medical problem. Results of this retrospective study showed the urgent need to pay more attention to the pretreatment examination of components of the oral microbiome, especially to the strains, which are etiological agents of human opportunistic infections and are particularly dangerous for older adults.

Comparative study on usefulness of gentamycin-containing collagen implants in the treatment of patients with osteitis and osteomyelitis of the craniofacial skeleton.

Introduction and objective. A reduction in incidences of peri-surgical complications due to infections is achieved by antibiotic prophylaxis The objective of the study was to assess the usefulness of gentamycin-containing collagen implants (GCCI) in the treatment of patients with osteitis and osteomyelitis of the craniofacial skeleton. Materials and method. The retrospective study included 103 patients with osteitis and osteomyelitis. 54 patients were treated intra-operatively with GCCI (Garamycin, EusaPharma, Europe). 49 patients were treated according to standard procedures. Light microscopy and in vitro culture techniques were applied for bacteria specific identification, and to investigate the resistance of detected microbiota to antibiotics. Patients received one dose of antibiotic pre-operatively. Post-operative antibiotic treatment was administered individually, according to clinical course and microbiological tests. The patients were followed-up on days 3, 7 and 14 after discharge for local complications; radiographic follow-up was performed 3, 6 and 12 months after surgery. Results. The course of post-operative antibiotic therapy was shorter in GCCI patients than in the control group (median 1 vs. 7 days); they also required shorter hospitalization (median 3 vs. 4 days). Implantation of GCCI significantly reduced the incidence of local complications (OR 0.30, 95%CI 0.11-0.83, p<0.0001), independently of the use of postoperative antibiotic therapy. On follow-up after 3-12 months, all patients presented with good soft tissue and bone healing. Conclusions. The results of this comparative study advocate the use of GCCI in osteomyelitis of various origin in oral and maxillofacial surgery, as they seemed to reduce the incidence of local complications, shorten antibiotic administration time and hospital stay.

Identification of infectious microbiota from oral cavity environment of various population group patients as a preventive approach to human health risk factors.

INTRODUCTION AND OBJECTIVE: This study presents the results of comparative investigations aimed to determine microbiota that can occur in the oral environment in different human populations. The objective of the research was to identify pathogenic oral microbiota, the potential cause of health complications in patients of different population groups. MATERIALS AND METHOD: The study included 95 patients requiring dental or surgical treatment; their oral cavity environment microbiota as risk factors of local and general infections were assessed. RESULTS: In clinical assessment, differences occurred in oral cavity conditions between patients with malformations of the masticatory system, kidney allograft recipients and individuals without indications for surgical procedures. The presence of various pathogenic and opportunistic bacterial strains in oral cavities were revealed by direct microscopic and in vitro culture techniques. CONCLUSIONS: Colonization of oral cavities of patients requiring surgical treatment by the potentially pathogenic bacteria constitutes the threat of their spread, and development of general infections. Assessment of oral cavity infectious microbiota should be performed as a preventive measure against peri-surgical complications.

Comparative assessment of selected intraoral microorganisms ­ potential factors for peri-surgical management complications

In this research, a comparative analysis of results of investigations involving different human populations, in terms of a relation between the oral cavity health and the species composition of mouth microbiota is reported. The purpose of this analysis was to identify and assess microorganisms that could cause health complications in patients with neoplasm requiring dental problem-related surgical management. The patients with the oral cancer surgically treated and those without neoplasm were assessed for their oral health: status of teeth, gingiva, periodontium, and occurrence of inflammatory processes. From each patient, microorganisms isolated of periodontium, dental plaque, and dental pocket swabs were identified in wet and stained microscopic preparations; standard microbiology in vitro techniques were also applied to determine the fungal and bacterial strains. The comparative analysis of results of direct microscopic examinations and in vitro cultures assessment indicated significant differences in prevalences of fungi, parasitic oral protozoans and bacteriae in particular patient's groups. Yeast-like fungi belonging to Candida genus, mostly of C. albicans group, were identified in 93.75% patients with the oral cancer, while in 25% of individuals assessed without neoplasm. E. gingivalis amoebae were only found in 12.5% patients with the serious disease; no trichomonads were detected in all patients analyzed. Among bacteria species, potentially pathogenic Enterobacteriaceae were found in the patients with oral cancer. The pronounced shift in the microbiota species composition in the patients who needed prolonged treatment due to oral cavity cancer, compared to other generally healthy persons has been showed in this analysis.

Evaluation of selected oral cavity microbiota--risk factors of management complications in patients with masticatory system disorders.

The retrospective analysis of data on oral cavity clinical status in relation to microbiota species composition is presented. The research regards patients of different age, with and without congenital malformation, pretreatment assessed for occurrence of pathological changes in the masticatory system. Samples of the swabs collected from each patient (from dental plaque, periodontium and dental pockets) were used for identification of oral protozoans in wet slides and stained preparations; additionally, transmission electron microscope examination was performed. The material was used for in vitro cultures to identify bacteria strains. Clinically, intensity of tissue deteriorations was higher in patients with a congenital disease. Alive Trichomonas tenax and Entamoeba gingivalis, species with confirmed pathogenic impact on oral cavity and neighboring structures, were detected with higher prevalence in older patients. Enterococci, Staphylococcus aureus, various Enterobacteriaceae were more frequently detected in patients with somatic and mental retardations; in mouths of those patients, Klebsiella pneumonia and Pseudomonas aeruginosa opportunistic strains occurred. Masticatory system abnormalities favor colonization of oral cavity by exogenous species and dissemination of infections, especially dangerous for patients with congenital diseases. Oral microbiota assessment and preventive measures may be helpful to avoid subsequent peri-surgery complications.

The threats for human health induced by food pests of Plodia interpunctella as reservoirs of infectious microbiota / Zagrozenia zdrowia czlowieka indukowane przez szkodniki zywnosci Plodia interpunctella jako rezerwuary infekcyjnych microbiota

INTRODUCTION: World-wide distributed pests of Plodia interpunctella occur with increasing frequency also in Poland, in areas where food is prepared and stored, in dwellings, buildings of public use, hospitals. Larvae damage various products causing economic losses. There were no data about microbiota transmission by pests. The aim of our systematic studies firstly conducted in Poland was to explain a role of pests as reservoirs of microbiota and assess health risk induced by them in human environments. MATERIAL AND METHODS: 300 adults and 200 larvae, collected in households and health facilities by traps and directly from products, were examined by light microscopy, in vitro cultivations, molecular techniques; the susceptibility /resistance of microbiota to chemicals was also assessed. RESULTS: Gram+ bacteriae of genera Enterococcus, Micrococcus, Bacillus, Gram-: Klebsiella, Escherichia, mold fungi: Aspergillus, Penicillium and yeast-like fungi were identified, including strains potentially pathogenic for humans. CONCLUSIONS: In the European Union countries, the food circulation is audited by the law; chemicals are applied to eliminate P.interpunctella pests causing economic losses. Our successive studies showed that pyralids may generate health problems as food pests and as reservoirs of microbiota. Sources of the pathogenic, drug-resistant strains revealed by us, not identified earlier, may be particularly dangerous for elder persons, with weakened immune system, persons from groups of high risk of infections. The increased awareness of the problem is necessary for more efficacy of preventive measures. A monitoring of consequences of the health risk induced by the pests may supply data useful for adequate practical approach.

Emerging Threats for Human Health in Poland: Pathogenic Isolates from Drug Resistant Acanthamoeba Keratitis Monitored in terms of Their In Vitro Dynamics and Temperature Adaptability.

Amphizoic amoebae generate a serious human health threat due to their pathogenic potential as facultative parasites, causative agents of vision-threatening Acanthamoeba keratitis (AK). Recently, AK incidences have been reported with increasing frequency worldwide, particularly in contact lens wearers. In our study, severe cases of AK in Poland and respective pathogenic isolates were assessed at clinical, morphological, and molecular levels. Misdiagnoses and the unsuccessful treatment in other ophthalmic units delayed suitable therapy, and resistance to applied chemicals resulted in severe courses and treatment difficulties. Molecular assessment indicated that all sequenced pathogenic corneal isolates deriving from Polish patients with AK examined by us showed 98-100% homology with Acanthamoeba genotype T4, the most prevalent genotype in this human ocular infection worldwide. In vitro assays revealed that the pathogenic strains are able to grow at elevated temperature and have a wide adaptive capability. This study is our subsequent in vitro investigation on pathogenic Acanthamoeba strains of AK originating from Polish patients. Further investigations designed to foster a better understanding of the factors leading to an increase of AK observed in the past years in Poland may help to prevent or at least better cope with future cases.