Search Results (785)

The demand for wild animal meat, popularly called “bushmeat”, serves as a driving force behind the emergence of infectious diseases, potentially transmitting a variety of pathogenic bacteria to humans through handling and consumption. This study investigated the microbial load and bacterial diversity in bushmeat sourced from a prominent bushmeat market in Kumasi, Ghana. Carcasses of 61 wild animals, including rodents (44), antelopes (14), and African civets (3), were sampled for microbiological analysis. These samples encompassed meat, intestines, and anal and oral swabs. The total aerobic bacteria plate count (TPC), Enterobacteriaceae count (EBC), and fungal counts were determined. Bacterial identification was conducted using MALDI-TOF biotyping. Fungal counts were the highest across all animal groups, with African civets having 11.8 ± 0.3 log₁₀ CFU/g and 11.9 ± 0.2 log₁₀ CFU/g in intestinal and meat samples, respectively. The highest total plate count (TPC) was observed in rodents, both in their intestines (10.9 ± 1.0 log₁₀ CFU/g) and meat (10.9 ± 1.9 log₁₀ CFU/g). In contrast, antelopes exhibited the lowest counts across all categories, particularly in EBC from intestinal samples (6.1 ± 1.5 log₁₀ CFU/g) and meat samples (5.6 ± 1.2 log₁₀ CFU/g). A comprehensive analysis yielded 524 bacterial isolates belonging to 20 genera, with Escherichia coli (18.1%) and Klebsiella spp. (15.5%) representing the most prevalent species. Notably, the detection of substantial microbial contamination in bushmeat underscores the imperative for a holistic One Health approach to enhance product quality and mitigate risks associated with its handling and consumption. Full article

Introduction/objectives: Peritonitis resulting from Candida spp. is common among critically ill patients and has been associated with adverse clinical outcomes. This study aimed to determine the effects of isolates of Candida species in patients with peritonitis on in-hospital mortality, general hospital stay, and intensive care unit (ICU) stays. Methods: This retrospective cohort study was conducted in two highly complex hospitals in Bogotá, Colombia, specifically by reference to patients who were hospitalized in the ICU between 2016 and 2022 with a clinical and microbiological diagnosis of peritonitis. For the analysis conducted for this research, two groups were established: patients with isolates of Candida spp. in the peritoneum and patients who had at least one bacterial microorganism in the culture. Multivariate logistic regression models and counting models featuring different mortality outcomes, different lengths of stay in the ICU, and different lengths of stay in the hospital were generated to evaluate the effect of the presence of Candida spp. and to account for potentially confounding variables. Results: A total of 373 patients, including 83 with Candida spp. and 290 with a bacterial etiology, were identified. Among the former group of patients, the most frequently identified species were C. albicans (50, 60.2%), C. tropicalis (18, 21.7%), and C. glabrata (7, 8.4%), whereas among the latter group, E. coli (186, 48.5%), K. pneumoniae (110, 29.8%), and E. faecalis (63, 16.9%) were most frequent. The 30-day mortality rate among patients with peritonitis and Candida isolates was 36.1%, and the corresponding rate among patients in the bacterial peritonitis group was 31.4% (p = 0.071). After adjustments were made to account for covariates, no significant differences were observed in mortality at 30 days (OR 0.75, 95% CI 0.20–1.18), length of hospital stay (iRR 1.11, 95% CI 0.90–1.40), or length of stay in the ICU (iRR 1.11, 95% CI 0.39) with respect to patients with peritonitis without fungal isolates. The Simplified Acute Physiology Score (SAPS2) (OR 1.04, 95% CI 1.03–1.06), World Society of Emergency Surgery (WSES) score (OR 1.11, (1.03–1.19), previous use of antifungals (OR 2.33, 1.21–4.52), and connective tissue disease (OR 3.71, 95% CI 1.30–10.99) were associated with 30-day mortality. Conclusions: The isolation of Candida species in peritoneal fluid from critically ill patients with peritonitis was not significantly associated with in-hospital mortality, length of hospital stay, or length of ICU stay after adjustments were made to account for other variables. Full article

Background/Objectives: Romania remains a tuberculosis (TB) hotspot in the European Union, yet host-derived factors of poor outcomes are poorly characterised. We quantified circulating pro-inflammatory cytokines and examined their interplay with behavioural risk factors, the nutritional status, and the clinical course in adults hospitalised with pulmonary TB. We analysed 80 adults with microbiologically confirmed pulmonary TB and 40 respiratory symptom controls; four TB patients (5%) died during hospitalisation, all within 10 days of admission. Methods: A retrospective analytical case–control study was conducted at the Constanța regional TB referral centre (October 2020—October 2023). Patients with smear- or culture-confirmed TB were frequency-matched by sex, 10-year age band, and BMI class to culture-negative respiratory controls at a 2:1 ratio. The patients’ serum interferon-γ (IFN-γ), interleukin-1α (IL-1α), interleukin-1β (IL-1β), and tumour-necrosis-factor-α (TNF-α) were quantified within 24 h of admission; the neutrophil/lymphocyte ratio (NLR) was extracted from full blood counts. Independent predictors of in-hospital mortality were identified by multivariable logistic regression; factors associated with the length of stay (LOS) were modelled with quasi-Poisson regression. Results: The median TNF-α (24.1 pg mL⁻¹ vs. 16.2 pg mL⁻¹; p = 0.009) and IL-1β (5.34 pg mL⁻¹ vs. 3.67 pg mL⁻¹; p = 0.008) were significantly higher in the TB cases than in controls. TNF-α was strongly correlated with IL-1β (ρ = 0.80; p < 0.001), while NLR showed weak concordance with multiplex cytokine patterns. Among the patients with TB, four early deaths (5%) exhibited a tripling of TNF-α (71.4 pg mL⁻¹) and a doubling of NLR (7.8) compared with the survivors. Each 10 pg mL⁻¹ rise in TNF-α independently increased the odds of in-hospital death by 1.8-fold (95% CI 1.1–3.0; p = 0.02). The LOS (median 29 days) was unrelated to the smoking, alcohol, or comorbidity load, but varied across BMI strata: underweight, 27 days; normal weight, 30 days; overweight, 23 days (Kruskal–Wallis p = 0.03). In a multivariable analysis, under-nutrition (BMI < 18.5 kg m⁻²) prolonged the LOS by 19% (IRR 1.19; 95% CI 1.05–1.34; p = 0.004) independently of the disease severity. Conclusions: A hyper-TNF-α/IL-1β systemic signature correlates with early mortality in Romanian pulmonary TB, while under-nutrition is the dominant modifiable determinant of prolonged hospitalisation. Admission algorithms that pair rapid TNF-α testing with systematic nutritional assessment could enable targeted host-directed therapy trials and optimise bed utilisation in high-burden settings. Full article

Accurate and efficient estimation of microalgae cell concentration is critical for applications in hydrochemical monitoring, biofuel production, pharmaceuticals, and ecological studies. Traditional methods, such as manual counting with a hemocytometer, are time-consuming and prone to human error, while automated systems are often costly and require extensive training data. This paper presents a low-cost, automated approach for estimating cell concentration in Chlorella vulgaris suspensions using classical computer vision techniques. The proposed method eliminates the need for deep learning by leveraging the Hough circle transform to detect and count cells in microscope images, combined with a conversion factor to translate pixel measurements into metric units for direct concentration calculation (cells/mL). Validation against manual hemocytometer counts demonstrated strong agreement, with a Pearson correlation coefficient of 0.96 and a mean percentage difference of 17.96%. The system achieves rapid processing (under 30 s per image) and offers interpretability, allowing specialists to verify results visually. Key advantages include affordability, minimal hardware requirements, and adaptability to other microbiological applications. Limitations, such as sensitivity to cell clumping and impurities, are discussed. This work provides a practical, accessible solution for laboratories lacking expensive automated equipment, bridging the gap between manual methods and high-end technologies. Full article

Objective: To evaluate the effect of a probiotic combination on clinical and oral microbiological parameters in patients with head and neck cancer (HNC) undergoing radiotherapy. Materials and Methods: A randomised, double-blind, placebo-controlled clinical trial was conducted with 72 HNC patients who had received radiotherapy within the past year. Participants were randomly assigned to receive either daily probiotic sachets or placebo for 30 days. Salivary parameters—including unstimulated and stimulated flow rates and pH—were evaluated alongside oral microbiota profiles, including total bacterial load and selected periodontopathogens. Assessments were performed at baseline and post-intervention using sialometry, pH analysis, bacterial culture, and quantitative real-time PCR (qPCR). Results: Sixty-one patients completed the study (31 in the probiotic group, 30 in the placebo group). Stimulated salivary flow increased significantly in the probiotic group (p = 0.0016), while unstimulated flow improved in both groups (p < 0.05). Salivary pH decreased significantly in the probiotic group (p = 0.0209); however, no intergroup differences were observed at the end of the intervention (p = 0.9839). qPCR showed significant reductions in total bacterial load (p = 0.0209) and Fusobacterium nucleatum (p = 0.0080). Culture confirmed the reduction of F. nucleatum (p = 0.0026), with a trend towards significance for total cultivable bacterial count (p = 0.0502). Conclusions: Daily supplementation with a probiotic combination may serve as a practical and well-tolerated adjunctive measure in clinical settings to improve salivary function and reduce key oral pathogens, particularly Fusobacterium nucleatum, in patients undergoing or recovering from radiotherapy for head and neck cancer. These findings support its potential integration into routine supportive care protocols to mitigate xerostomia and oral dysbiosis in this population. Full article

This study aimed to develop a fermented mango-based beverage using Lactiplantibacillus plantarum strains Lp6 and Lp32, focusing on enhancing its functional properties, ensuring microbiological safety, improving nutritional value, and achieving sensory acceptability. A central composite design (CCD) was employed to assess the effects of two factors (fermentation time and inoculum concentration) on several response variables: viable cell concentration (CC), total phenolic compounds (TPCs), total flavonoid compounds (TFCs), and concentrations of L-lactic acid and D-lactic acid. The optimized formulation was achieved using L. plantarum Lp6, with an inoculum concentration of 9.89 Log (7.76 × 10⁹) CFU/mL and a fermentation time of 20.47 h. Under these conditions, the beverage reached the highest values for CC, TPC, TF, and L-lactic acid while minimizing the production of D-lactic acid. Following optimization, the fermented beverage underwent further characterization, including physicochemical analysis, microbiological evaluation, proximate composition analysis, and sensory evaluation. The final product exhibited a viable cell count of 13.01 Log (10.23 × 10¹²) CFU/mL, demonstrated functional potential, complied with microbiological safety standards, and showed adequate nutritional content. Sensory analysis revealed high consumer acceptability, attributed to its distinctive mango aroma and flavor. These findings highlight the potential of this fermented mango-based beverage as a novel functional food with promising market appeal. Full article

Background: Monolithic zirconia has attracted considerable interest in dentistry due to its favorable physical and mechanical properties, making it a promising alternative for crown fabrication. Nonetheless, a standardized finishing protocol for this material has yet to be established. Objective: This study aimed to evaluate the surface characteristics and in vitro biofilm formation of zirconia finished by either polishing or glazing. Methods: A total of 72 zirconia specimens were fabricated and divided into control, glazing, and polishing groups. Surface analysis included roughness, wettability, and surface free energy. Microbiological analysis included CFU (colony-forming units per mL) counts, microbial adhesion at 2, 4, 6, and 8 h, biofilm biovolume, and qualitative biofilm assessment via scanning electron microscopy (sEm). Results: The glazing group showed significantly greater roughness than the polishing (p = 0.006) and control (p = 0.016) groups, along with a lower contact angle (polishing—p = 0.002; control—p < 0.001) and higher surface energy (polishing—p = 0.005; control—p < 0.001). No significant differences were observed in CFU counts for the tested microorganisms (C. albicans, p = 0.158; L. casei, p = 0.610; S. mutans, p = 0.904). Regarding microbial adhesion, the polishing group showed a smaller biofilm-covered area compared to the control group for both total biofilm (p = 0.008) and viable biofilm (p = 0.005). no statistically significant difference was observed in biofilm biovolume (p = 0.082). Conclusions: These findings suggest that, despite the surface differences among the groups, biofilm formation was not significantly affected. Full article

Objectives: Cefideroccol (FDC) is a siderophore cephalosporin with potent antibacterial activity against a wide range of Gram-negative multidrug-resistant (MDR) microorganisms. We investigated the anti-biofilm capacity of FDC against clinical strains. Methods: This multicenter study was conducted on 28 selected strains of MDR Gram-negative bacilli isolated from clinical samples of Pseudomonas aeruginosa (n = 5), Acinetobacter baumannii (n = 11), and Klebsiella pneumoniae (n = 12). We first determined the minimum inhibitory concentration (MIC) of each strain using the microdilution method. We also defined the minimum biofilm inhibitory concentration (MBIC) as a ≥50% reduction in tetrazolium salt (XTT) (as recommended in the 2017 Spanish Microbiology Protocols [SEIMC] for the microbiological diagnosis of infections related to the formation of biofilms). We also analyzed the reduction in the following biofilm variables after an 8 mg/mL FDC treatment: the CFU count, the cell viability, the biomass, the metabolic activity, and extracellular α or β polysaccharides. Results: The MIC₅₀ and MBIC₅₀ of FDC were 0.5 mg/L and 64 mg/L, respectively. We observed a mean (SD) fold increase in the susceptibility to FDC between planktonic and sessile cells for P. aeruginosa, A. baumannii, and K. pneumoniae of 9.60 (0.55), 6.27 (2.28), and 6.25 (2.80), respectively. When 8 mg/mL of FDC was tested, we observed that the best median (IQR) percentage reductions were obtained for cell viability and the extracellular matrix (73.1 [12.4–86.5] and 79.5 [37.3–95.5], respectively), particularly for P. aeruginosa. The lowest percentage reduction rates were those obtained for biomass. Conclusions: We demonstrated that the susceptibility to FDC was significantly reduced when strains were in a biofilm state. The best percentage reduction rates for all biofilm-defining variables were observed for P. aeruginosa. Our results need to be validated using a larger collection of clinical samples. Full article

Micro-fragmented adipose tissue (MFAT) is a promising autologous therapy for knee osteoarthritis. To avoid repeated liposuction procedures for its clinical application, MFAT obtained from patients with knee osteoarthritis was stored at −80 °C in a tissue bank. This study describes the preparation, cryopreservation, thawing, and washing, as well as comprehensive analysis of cell populations in fresh and MFAT thawed after two years. Immunophenotyping of both fresh and thawed MFAT showed a significant presence of endothelial progenitors and pericytes in the stromal vascular fraction. Viability before (59.75%) and after freezing (55.73%) showed no significant difference. However, the average cell count per gram of MFAT was significantly reduced in thawed samples (3.00 × 10⁵) compared to fresh ones (5.64 × 10⁵), likely due to processing steps. Thawed MFAT samples showed increased CD73 expression on the CD31^highCD34^high subset of EP and SA-ASC, as well as increased expression of CD105 on EP, the CD31^lowCD34^low subset of EP, pericytes, and SA-ASC. Microbiological testing confirmed 100% sterility, and double washing efficiently removed DMSO, confirming sample safety. Histological analysis revealed healthy, uniformly shaped adipocytes with intact membranes. This approach allows accurate estimation of cell yield for intra-articular injection, ensuring delivery of the target cell number into the knee. Quality control analysis confirms that cryopreserved MFAT retains high cellular and structural integrity, supporting its safety and suitability for clinical application. Full article

This study analyzes the aromatic profiles of kiwi-based fermented beverages, inoculated with varying proportions of milk kefir grains and incubated under different shaking rates. The experiments were designed using response surface methodology and three consecutive batch cultures were performed under each experimental condition. At the end of each fermentation, the grains were separated from the beverage and reused as the inoculum for fermenting fresh kiwi juice in the subsequent batch. Based on the results, together with the previously determined microbiological and chemical characteristics, two beverages were identified as having broader aromatic profiles, lower contents of sugars, ethanol, and acids, and high counts of lactic acid bacteria (LAB) and yeasts (>10⁶ CFU/mL). These beverages were produced under relatively low agitation rates (38 and 86 rpm) and high inoculum proportions (4.33% and 4.68% w/v) during the second and third batch cultures, respectively. Over 28 days of refrigerated storage, the pH values of both beverages remained relatively stable, and the LAB counts consistently exceeded 10⁶ CFU/mL. Yeast counts, along with the production of ethanol, glycerol, lactic acid, and acetic acid, increased slightly over time. In contrast, the concentrations of citric acid, quinic acid, total sugars, and acetic acid bacteria declined by day 28. Full article

Objectives: Carbapenem-resistant Gram-negative bacteria (CR-GNB) infections in intensive care units (ICUs) are increasingly prevalent and associated with high mortality. This study aimed to investigate the distribution of isolated bacteria and determine the factors associated with mortality among ICU patients diagnosed with CR-GNB infections. Methods: This retrospective study included 95 patients admitted to the ICU between February 2022 and July 2024 who were diagnosed with CR-GNB infections via culture and initiated on treatment. Thirty-day mortality was defined as the clinical outcome, and patients were divided into two groups: survivors (Group 1, n = 42) and deceased (Group 2, n = 53). Demographic, clinical, laboratory, and microbiological data were analyzed. Results: Advanced age, the presence of malignancy, an elevated Charlson Comorbidity Index (CCI), lower platelet counts, and higher C-reactive protein (CRP) levels were significantly associated with mortality (p < 0.05). Trauma-related admissions were more common among survivors, while sepsis-related admissions predominated among non-survivors. No statistically significant associations were observed between antibiotic regimen type and mortality. Culture-based pathogen distribution revealed A. baumannii as the predominant organism in respiratory samples, while K. pneumoniae was more frequently isolated from bloodstream and urinary specimens. Conclusions: Mortality in ICU patients with CR-GNB infections is influenced by both baseline comorbidities and infection-related inflammatory markers. This study provides region-specific insights from a high-resistance ICU setting and may inform risk stratification, prognostication, and management strategies in critically ill patients with CR-GNB infections. Full article

Background: Zoonotic infection with Mycobacterium bovis continues to occur, particularly in regions lacking bovine tuberculosis surveillance and where the consumption of unpasteurized dairy products, including artisanal cheeses, is common. We describe the clinical and microbiological characteristics, diagnostic procedures, and treatment outcomes of individuals with HIV with M. bovis infection. Methods: We conducted a retrospective study analyzing sociodemographic, clinical, microbiological, and computed tomography (CT) data, as well as treatment outcomes, in 12 patients with HIV with confirmed M. bovis infection. These findings were compared with those of 14 individuals with HIV diagnosed with Mycobacterium tuberculosis infection during the same period. Results: Consumption of unpasteurized dairy products was significantly associated with M. bovis. Patients with M. bovis infection had higher CD4+ T-cell counts compared to those with M. tuberculosis infection (p = 0.01, r = 0.45). All M. bovis cases presented with extrapulmonary disease. CT imaging in M. bovis infection more frequently demonstrated retroperitoneal lymphadenopathy, hepatosplenomegaly, and splenic abscesses compared to M. tuberculosis infection. Microbiological identification was exclusively from extrapulmonary sites in all M. bovis cases. Surgical interventions, including abscess drainage or splenectomy, were significantly more common among M. bovis patients. Conclusions: M. bovis infection in individuals with HIV is characterized by consistent extrapulmonary, often abdominal, involvement. Surgical procedures are frequently required for both diagnosis and management. Targeted efforts to identify M. bovis are warranted, particularly in high-burden regions where unpasteurized dairy consumption remains prevalent. Full article

Insulin-like growth factor-binding protein-3 (IGFBP-3) plays a vital role in cellular growth, development, and survival. Incorporating IGFBP-3 into baseline prognostic evaluations may enhance the prediction of mortality in patients with sepsis. In this study, serum levels of IGFBP-3, C-reactive protein, procalcitonin, lactate, interleukin-6, and mid-regional pro-adrenomedullin were measured upon admission to the internal medicine unit (IMU) in 139 patients with microbiologically confirmed sepsis. The objectives were as follows: (1) to classify septic patient phenotypes based on optimal thresholds of independent prognostic biomarkers and (2) to evaluate whether these biomarkers improve the predictive accuracy of a clinical model (Model 1), which includes the clinical predictors of 1-year mortality. Age, sequential organ failure assessment (SOFA) score, multiple sources of infection, and IGFBP-3 levels independently predicted 1-year mortality. Patients with IGFBP-3 levels below 10.64 had significantly lower median body temperature (p = 0.008), reduced lymphocyte count (p = 0.001), and higher 1-year mortality (p < 0.001). Model 1 included age, SOFA score, and the presence of multiple sources of sepsis as predictor variables. Model 2 incorporated the same variables as Model 1, with the addition of IGFBP-3 levels. When comparing their prognostic performance, Model 2 demonstrated superior predictive accuracy for mortality at 60, 90, and 365 days following admission to the IMU. Low IGFBP-3 levels at IMU admission are strongly associated with worse outcomes in septic patients, supporting its potential use as a prognostic biomarker. Full article

Sixteen long-ripened, high-quality Cabrales cheeses from independent producers underwent a comprehensive biochemical and microbiological characterisation. Significant variations in total microbial counts and specific microbial groups were observed among the cheeses. A metataxonomic analysis identified 249 prokaryotic amplicon sequence variants (ASVs) and 99 eukaryotic ASVs, respectively, which were classified into 52 prokaryotic and 43 eukaryotic species. The predominant species included bacteria of the genera Tetragenococcus, Lactococcus (of which Lactococcus lactis was used as a starter), and Staphylococcus, followed by Brevibacterium and Corynebacterium species. The starter mould Penicillium roqueforti was highly abundant in all cheeses; Debaryomyces hansenii, Geotrichum candidum, and Kluyveromyces spp. constituted the subdominant fungal populations. Glutamic acid (≈20 mg g⁻¹) was the most abundant free amino acid in all samples, followed by lysine, leucine, and valine (≈10–13 mg g⁻¹). Moderate-to-high amounts of the biogenic amines tyramine and ornithine were detected. A large variation between cheeses of the main organic acids (lactic, acetic, or butyric) was detected. Differences between samples were also observed for the majority volatile compounds, which included organic acids, alcohols, esters, and ketones. Positive and negative correlations between bacterial and fungal species were detected, as well as between microbial populations and key biochemical markers. Among the latter, Tetragenococcus halophilus correlated positively with ethyl caprylate and hexanoic acid, and Loigolactobacillus rennini correlated positively with γ-aminobutyric acid. Conversely, Staphylococcus equorum showed a strong negative correlation with ethyl caprylate and capric acid. These microbial and biochemical insights enabled us to propose a microbiota-based starter culture comprising prokaryotic and eukaryotic components to enhance Cabrales cheese quality. Full article

Edible insects are gaining traction worldwide; however, the existing data regarding their microbiological quality remain inadequate. This study investigated the bacterial communities and microbiological quality of five types of frozen edible insects commercially available in Thailand. Amplicon sequencing revealed Firmicutes (Bacillota) and Proteobacteria (Pseudomonadota) as the main phyla across all samples; Bacteroidota was predominant in house crickets, Actinobacteriota in silkworms, and Desulfobacterota was exclusively found in house and mole crickets. Culture-based assays showed total viable counts, lactic acid bacteria, yeasts–molds, and spore-formers ranging from 3.41–6.58, 2.52–7.41, 1.83–5.62, to 2.00–4.70 log CFU·g⁻¹, respectively. In some samples, Enterobacteriaceae and Escherichia coli, key hygiene indicators, reached 5.05 and 2.70 log CFU·g⁻¹, respectively. Among foodborne pathogens, presumptive Bacillus cereus was found to vary from <1.70 to 3.93 log CFU·g⁻¹, while Clostridium perfringens and Staphylococcus aureus were under the quantitation limit, and Salmonella was absent. Overall, the results indicate significant variation in microbial diversity and quality among different insect types. The high levels of microbial hygiene indicators and foodborne pathogens in some samples raised food safety concerns and point to the need to develop or implement production guidelines and microbiological criteria for frozen edible insects to ensure food safety. Full article