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Introduction: Shiga toxin-producing Escherichia coli (STEC), particularly E. coli O157:H7, is a major contributor to foodborne outbreaks globally. Both E. coli O157 and non-O157 strains can lead to severe health issues, including hemolytic colitis and hemolytic uremic syndrome, which can result in kidney failure.

Methods: Two hundred and one frozen beef samples were purchased from various supermarkets located in the Eastern Province of Saudi Arabia and subsequently enriched in tryptic soy broth (TSB). From the enriched samples in TSB, 1 mL portion was mixed with immunomagnetic beads (IMB) coated with specific antibodies targeting the E. coli O157 O antigen. The beads, which contained the captured bacteria, were then streaked onto CHROMagar O157 and Sorbitol MacConkey (SMAC) agar. The DNA extracted from these samples was examined using multiplex PCR to identify potential virulence gene markers, specifically stx-1, stx-2, and eae.

Results: Of the 201 examined samples, 88 (43.8%) and 106 (52.7%) were positive for E. coli and produced colorless and mauve colonies on SMAC agar and CHROMagar O157, respectively. Out of 298 isolates in total, 174 isolates of E. coli were isolated with IMB enrichment. The highest detection rate of virulence gene markers was found among isolates that had been isolated using IMB enrichment, where 25 (8.4%), 2 (0.7%) and 12 (4%) isolates tested positive for stx1, stx2, and eae genes respectively. Among 42 isolates harboring potential virulence gene markers, 11 isolates were identified as E. coli O157 (stx ₁ ⁺/eae ⁺ or stx ₂ ⁺/eae ⁺). ERIC-PCR genotyping was able to determine the genetic relatedness among 42 isolates of E. coli O157 and E. coli non-O157 into 10 types with four identical related clusters and a genetic similarity rate above 90% homology from the identified isolates.

Conclusions: The present study gives a clear perspective on STEC contamination in imported frozen beef marketed in Saudi Arabia. Because of the many possibilities of STEC contamination in imported frozen beef, further studies on the spread of STEC at various levels of imported frozen meat are needed on a long-term basis.

Many microorganisms are used to produce antibiotics, vaccines, and medicines for various diseases, and preserving these microorganisms and viruses can ensure that the development process is streamlined. We have reviewed the short-term and long-term microbial and viral preservation methods including novel and emerging technologies. Short-term preservation methods of microorganisms are designed to maintain the viability of the organisms for periods ranging from a few days up to one year. The novel microfluid systems allow for the creation of microenvironments that support the growth and proliferation of specific microorganisms and the isolation of individual cells or small populations of microorganisms for studying microbial behavior and interactions. Long-term preservation involves storing the organisms for an extended period, ranging from months to decades, while retaining their viability and genetic stability. The mineral oil or liquid paraffin storage, storage in distilled water, storage in sterile soil, lyophilization, and cryopreservation are well known, and encapsulation of nanoparticles to preserve microorganisms, electrospinning, and electro spraying and supercooling are novel and emerging methods. Each short and long-term microbial and viral preservation method has advantages and disadvantages, and, based on the requirement, the appropriate method can be chosen.

Introduction: The emergence of colistin resistance threatens the treatment of Pseudomonas aeruginosa infections.

Methods: In this study, in vitro development of colistin resistance was investigated using comparative phenotypic and proteomic analysis of P. aeruginosa ATCC 9027, its 14-day colistin sub-MIC exposed strain (Col-E1), and 10-day antibiotic-free cultured Col-E1 strain (Col-E2). Antibiotic susceptibility, morphology, virulence factors, and proteomic changes were assessed using disc-diffusion, agar-based, spectrophotometry, SEM, and iTRAQ-LC-MS/MS methods.

Results: Colistin-exposed strains decreased susceptibility to colistin while remaining susceptible to other antibiotics. Col-E1 reduced the cell lengths by 17.67% and the colony size by 36.16% compared to the initial strain. The reduction remained in Col-E2. The pyocyanin production was reduced in Col-E1 (p=0.025, Tukey HSD) and increased again in Col-E2 (p=0.005, Tukey HSD). In contrast, no significant changes in elastase, protease, rhamnolipid, pyoverdine, and biofilm production were observed (p>0.05, Tukey HSD). In Col-E1, the proteome analysis showed 135 differentially expressed proteins (DEPs) of which 94 DEPs (69.23%) maintained their expression change in Col-E2. Among DEPs, 82 were involved in metabolism and protein synthesis. Some DEPs (6/135) played a role in stress response such as GrpE (fold change: 14.93) and Hmp (fold change: 12.08). In particular, membrane proteins like OprD, DdlB, and OprI showed significant colistin response with fold change of -8.47, 6.43 and 6.19, respectively.

Conclusions: In summary, colistin response in P. aeruginosa seemed to affect morphology, production of pyocyanin, and proteins of metabolism, protein synthesis, stress response and membrane.

Introduction: Central nervous system (CNS) infection due to the varicella zoster virus (VZV) can complicate the primary infection or the reactivation, leading to significant mortality and morbidity. This study aimed to describe the clinical, laboratory, and radiological characteristics of patients with confirmed VZV CNS infection in a tertiary hospital in Greece.

Methods: Data about patients hospitalized from January 2018 to September 2023 with CNS infection by VZV, confirmed by a syndromic polymerase chain reaction in the cerebrospinal fluid (CSF), were retrospectively collected and evaluated.

Results: In total, 14 patients were recorded. The median age was 49 years, and 35.7% were male. Headache was present in 71.4%, a rash in 57.1%, and fevers, nausea or vomiting, and disorientation in 35.7%. The CSF showed lymphocytic pleocytosis in all patients. Brain magnetic resonance imaging was performed in 53.8%. Empirical antivirals were given in 69.2%, and intravenous acyclovir was given to all patients after identification of VZV in the CSF. Only 7.1% (1 patient) required intensive care unit admission, and 7.1% (1 patient) died. Patients presenting without a rash may be slightly younger, have a slightly lower Charlson comorbidity index, be more likely to present with photophobia or phonophobia, and have lower serum CRP.

Conclusions: Patients presenting with VZV CNS infection have lymphocytic pleocytosis in the CSF and usually have a favorable outcome with antiviral treatment. Those presenting without a rash may have a different overall clinical phenotype from those with a rash; however, this must be evaluated in larger studies in the future.

Introduction: Dengue infection poses a serious threat to global public health, including Indonesia. The rapid spread and significant economic impact are crucial concerns for control efforts. Investigating risk factors of dengue virus infection is necessary to formulate effective strategies, particularly at the household level. This study aims to investigate contributing risk factors to dengue virus transmission in the Borobudur Subdistrict, Magelang, Indonesia, an area with persistently high dengue infection mortality rates.

Methods: This study adopted a case-control design and utilized secondary data collected from six villages in the Borobudur Subdistrict. A total of 111 households (37 cases and 74 controls) participated in the study and completed a questionnaire encompassing: 1) Sociodemographic data; 2) Healthy housing components; 3) Sanitation and behavioral components; and 4) Dengue infection prevention practices. Data were analyzed bivariately using statistical software to identify differences in group proportions.

Results: The findings of this study indicate that the presence of stagnant water (odds ratio [OR]: 5.02) and mosquito larvae (OR: 4.80) around the house, morning sleep habits (OR: 6.97), and lack of participation in anti-dengue programs (OR: 3.23) are significant risk factors (p<0.05) for dengue infection. However, no significant differences (p>0.05) were found in healthy housing components between the case and control groups.

Conclusions: This study has identified contributing risk factors to dengue virus transmission in the Borobudur Subdistrict. These results can serve as a foundation for designing more effective intervention programs for dengue infection and future prevention efforts.

High quality research is critical for evidence-based decision making in public health and fundamental to maintain progress and trust in immunization programs in Europe. In 2024 the European Centre for Disease Prevention and Control (ECDC) conducted an update of the 2020 systematic review to capture more recent evidence on of the efficacy, effectiveness of influenza vaccines in individuals aged 18 years and older in the prevention of laboratory-confirmed influenza. While this report was highly anticipated due to the strength of the protocol and processes put in place, during our assessment, we expressed two chief concerns. We are concerned by the grading of the evidence certainty applied and being unable to reproduce some data extracted in the report from the primary sources. While the systematic review benefited of strong methods and processes, the execution of the research protocol warrants revision due to the issues discussed. We encourage the ECDC to work towards an updated review within a reasonable time frame to avoid misinterpretation by decision-making bodies across Europe.

Introduction: The COVID-19 pandemic has promoted an intensive investigation into the pathophysiological mechanisms of SARS-CoV-2 infection, risk factors, and its impact on disease severity. Vitamin D has generated significant attention for its potential role in viral prevention and immune defense due to its pleiotropic functions, including immunomodulation and antimicrobial effects. This study aimed to assess serum 25(OH)D3 levels in patients with COVID-19 compared to those with other viral respiratory infections and to evaluate associations of vitamin D levels with symptomatology, clinical characteristics, presence of comorbidities and laboratory investigation.

Methods: The study included 78 patients admitted to a hospital with COVID-19 (52 patients) or other viral respiratory infections (26 patients). Routine blood biomarkers, markers of inflammation, markers of endothelial dysfunction, serum 25(OH)D3 were analyzed, and patients were classified according to vitamin D levels and presence of comorbidities.

Results: Most patients had vitamin D levels <30 ng/mL and there was no significant difference in 25(OH)D3 levels between patients with and without COVID-19 (p=0.768). Aging and comorbidity prevalence were significantly increased in the COVID-19 than in the non-COVID-19 group (p<0.001; p=0.049). A significant positive correlation was determined between endocan level and serum ferritin concentration in patients with COVID-19 and vitamin D deficiency. A borderline significantly elevated NLR was observed in patients with COVID-19 who were also vitamin D deficient, compared with the similar non-COVID-19 subgroup (p=0.05). In patients with COVID-19 and insufficient vitamin D, levels of 25(OH)D negatively correlated with endocan. Interestingly, COVID-19 patients with diabetes exhibited significantly lower 25(OH)D3 levels compared to non-diabetic patients (p=0.003), along with higher ferritin levels, suggesting a potential association between vitamin D deficiency and diabetes in COVID-19.

Conclusions: These findings contribute to the understanding of the complex interplay between vitamin D status, comorbidities, and COVID-19 outcomes, emphasizing the need for further research to elucidate their underlying mechanisms and clinical implications.

Introduction: Alveolar echinococcosis is one of the most pathogenic zoonoses caused by the larval forms of Echinococcus multilocularis. It is endemic in central Europe, but from 2001 to 2018, eight European countries reported their first cases of alveolar echinococcosis. These numbers testify to unprecedented spread of the infection.

Case report: We report the first case of alveolar echinococcosis from southern Croatia that was incidentally found in an immunocompromised 70-year-old male patient. He was admitted to the hospital due to macrohematuria and renal insufficiency. Diagnostic assessment of kidney impairment revealed a large liver infiltration whose radiographic appearance was suspicious of a cancer. Nevertheless, histopathological and molecular analysis of the liver biopsy confirmed alveolar echinococcosis. The lesion was irresectable due to expansion along the major liver vessels. Therefore, conservative treatment with albendazole was started and the existing immunosuppressive therapy was modulated. The control imaging following 11 months of albendazole treatment showed regression of the lesions.

Conclusions: It is not clear whether this case was imported or autochthonous, but it is evident that the incidence of alveolar echinococcosis is rising in Europe. Due to its rareness and malignant nature of the disease, it is often misdiagnosed or diagnosed late when radical surgical treatment is impossible to perform. Clinicians should be aware of this emerging parasitic disease, especially in immunocompromised patients, because every delay in reaching the diagnosis seriously hampers therapeutic efficacy. Better therapeutic options and standardized guidelines on the modulation of immunosuppressive therapy in these patients are highly needed.

Introduction: This paper examines the use of local antibiotic therapy in one-stage septic revision surgery for late periprosthetic joint infections (PJIs). This case study suggests that morselized bone allografts impregnated with antibiotics in powder form are a preferable alternative to polymethyl methacrylate (PMMA) because they can generate higher local antibiotic concentrations. Current research also recommends using vancomycin and aminoglycosides as the preferred choice of antibiotics, as they may have low diffusion in tissues when administered intravenously, but are effective when administered locally. The article emphasizes the importance of achieving high local antibiotic concentrations to eradicate bacterial biofilms and provides guidelines for the preparation of bone allografts.

Case report: The paper assesses the case study of a 68-year-old male patient who underwent two-stage total revision surgery for a late septic failure of the endoprosthesis (approximatively one year after implantation). The first stage involved removing the implant, debridement, lavage, and setting a fixed spacer manually made from polymethyl methacrylate impregnated with 4 g of vancomycin. The second stage of revision surgery utilized a morselized bone allograft impregnated with 4 g of vancomycin as a means of local antibiotic therapy and bone defect coverage. Systemic levels of vancomycin were measured at 4, 8, 12 and 24 hours respectively after surgery. During this period, no systemic vancomycin was administered. After the second stage was carried out and the testing was carried out, systemic vancomycin therapy was initiated and the plasma levels of the drug were assessed and subsequently therapeutic doses were adjusted.

Conclusions: Vancomycin loaded bone allograft achieved higher systemic concentrations that its polymethyl methacrylate counterpart, when being manually loaded with the same amount of antibiotic. Nonetheless, the levels were well below nephrotoxic levels, indicating that this may be a valuable tool for local means of antibiotic therapy in selected patients that could not endure such a systemic therapeutic regiment. For septic revisions, antibiotic loaded bone allograft holds a valuable place in the surgical arsenal of local antimicrobial treatment, by far exceeding that of the polymethyl methacrylate.

Introduction: Aggregatibacter (Actinobacillus) actinomycetemcomitans is a commensal bacterial pathogen in the human oral cavity. It can, however, represent the source of local or systemic infections with serious evolution, in particular infective endocarditis. We present a particular case of an adult male patient with infective endocarditis with A. actinomycetemcomitans and patent ductus arteriosus (PDA).

Case report: A 37-year-old patient, chronic ethanol user, is hospitalized for altered general condition, persistent cough, left chest pain, headache and dizziness, symptoms evolving for about 3 weeks. The clinical examination revealed crackling pulmonary rales present basally bilaterally, as well as numerous cavities and dental abscesses. Chest radiography showed mixed left hiliobasal pneumonia. Chest CT depicted pulmonary abscess and two filling defects in the pulmonary artery trunk, possible thrombotic/vegetative images/mediastinal thrombotic/adenopathic images. Broad spectrum antibiotic treatment was initiated. Transthoracic ultrasonography visualized persistence of ductus arteriosus and an echodense formation attached to the lateral wall of the pulmonary artery trunk. Following positive blood cultures for Aggregatibacter actinomycetemcomitans, the diagnosis of infective endocarditis was established and antibiotic treatment was de-escalated to ceftriaxone according to the antibiogram. The clinical course under treatment was slowly favorable, the patient was discharged on request on day 44 with continued treatment at home.

Conclusions: Infective endocarditis caused by Aggregatibacter actinomycetemcomitans should be considered in patients with altered general condition and congenital cardiovascular defects. In the present case, the patient presented two risk factors, namely poor dental hygiene and PDA.