Search Results (26)

Background: Chronic rhinosinusitis (CRS) affects 5.5–28% of the population and is primarily an inflammatory disease, with microbiota potentially playing a key role. Understanding microbial pathogens and resistance patterns is crucial for effective management. This study aimed to evaluate the incidence of Gram-negative rods in CRS in adults as a part of a prospective microbiological study. Methods: Over one year, paranasal sinus mucosa samples from CRS patients and nasal concha samples from controls were analyzed. Cultivable bacterial flora was assessed using culture-based methods. Biofilm formation was evaluated via a microtiter-plate assay, and antibiotic susceptibility was tested using the disk diffusion method. Results: Tissue samples from 74 CRS patients and 47 controls yielded 198 bacterial strains. Gram-positive cocci dominated, while Gram-negative rods accounted for 17.6%, with Escherichia coli, Klebsiella oxytoca, and Citrobacter spp. being most common. All Gram-negative rods formed biofilms in vitro. They were susceptible to cefotaxime, aztreonam, ciprofloxacin, and meropenem but showed varying sensitivity to ampicillin (20–67%), tigecycline (40–57%), and amoxicillin/clavulanic acid (73–83%). Conclusions: The result of this study underlines that treatment of CRS should be based on the result of drug susceptibility testing of the isolated microorganism. Full article

This study examines temporal patterns in pathogens isolated from prosthetic joint infection (PJI) cases and antimicrobial resistance patterns at a Romanian orthopedic center. We have conducted a retrospective cohort study that included 674 patients undergoing hip or knee replacement revision surgery between January 2016 and December 2023. From these, 102 confirmed PJI cases requiring surgical intervention were selected for analysis. We isolated 27 microorganisms from acute PJI cultures and 82 from chronic PJIs. Staphylococcus epidermidis (33 cases, 30.3%; 95% CI 22.0–40.3) was the predominant pathogen, with coagulase-negative Staphylococci (22 cases, 20.18%; 95% CI 0.9–41.3) and Enterobacteriaceae (13 cases, 11.9%; 95% CI 6.4–18.3) also prevalent. Methicillin resistance was identified in 43.6% of coagulase-negative staphylococci and 45.5% of Staphylococcus aureus isolates. All Gram-positive isolates remained susceptible to vancomycin, linezolid, and tigecycline. Among Gram-negative bacilli, Klebsiella oxytoca and Proteus mirabilis showed resistance to third-generation cephalosporins, with phenotypic profiles suggestive of extended-spectrum β-lactamase (ESBL) production. All Escherichia coli, Enterobacter spp., and Citrobacter freundii strains were fully susceptible to tested agents, while Pseudomonas aeruginosa exhibited reduced susceptibility to ciprofloxacin, aztreonam, and imipenem. Among the isolated strains, 47 were multidrug-resistant (MDR), with Staphylococcus aureus accounting for the highest MDR count, including methicillin resistance. The distribution of microorganism types and MDR strains remained consistent throughout the study period, with no significant association between infection type and MDR strain presence or between infection site and microorganism presence except for a strong association between MDR strains and the type of microorganism (p < 0.05). The microbial profile and resistance patterns in PJIs have remained stable over eight years. Our observations do not suggest that MDR PJIs are more commonly acute cases, contrary to what has been highlighted in previous reports. The ongoing prevalence of MDR strains underscores the importance of targeted antimicrobial treatments based on local susceptibility profiles. Full article

Breastfeeding is fundamental for the development and protection of the newborn, and microorganisms present in breast milk are associated with the development of the infant’s intestinal microbiota. However, there are factors that interfere with breastfeeding, resulting in the need to supply donated milk to milk banks for these children. Even though there is a restriction on medications prescribed for pregnant and breastfeeding women, some antimicrobials are accepted, as long as they are used correctly and as they can increase the selection pressure for resistant bacteria. The microorganisms present in breast milk from a human milk bank were evaluated and the resistance of the isolates to antimicrobials was phenotypically characterized. In total, 184 microbial isolates were identified by mass spectrometry, of 12 bacterial genera and 1 yeast genus. There was a high prevalence of bacteria of the genus Staphylococcus, mainly S. epidermidis (33%). Resistance to antimicrobials varied among species, with a higher percentage of isolates resistant to penicillins and macrolides. Multidrug resistance was identified in 12.6% of 143 isolates. Breast milk contains a wide variety of microorganisms, mainly those of the Staphylococcus and Enterobacter genera. There was a high percentage of resistant isolates, and multidrug resistance in Klebsiella oxytoca (66.7%; 4/6) and S. epidermidis (15.0%; 9/60) isolates, which increases the public health concern. Full article

Fungal endophytes, symbiotic microorganisms residing within plants, are renowned for producing bioactive secondary metabolites with diverse beneficial properties. We investigated the antimicrobial potential of fungal endophytes isolated from Solanum mauritianum, an invasive weed, against clinically significant bacterial pathogens. Selected fungal endophytes (Penicillium chrysogenum, Fusarium sp., and Paracamarosporium leucadendri) were isolated from the plant’s leaves and fruits. Their crude extracts were tested against various referenced strains, such as Mycobacterium species (M. smegmatis ATCC 607 and M. bovis ATCC 27290), Staphylococcus aureus ATCC 6571, Bacillus subtilis ATCC 11774, Klebsiella species (K. pneumoniae ATCC 10031 and K. oxytoca ATCC 8724), Escherichia coli ATCC 10536, and Pseudomonas aeruginosa ATCC 10145, using the Kirby-Bauer disk diffusion method. Resazurin Microtiter Assay was used for the determination of the minimum inhibitory concentration. The chemical nature of the secondary metabolites in the crude extracts produced by fungal endophytes was evaluated using high-resolution liquid chromatography–mass spectrometry (LC-MS) using water and acetonitrile gradient. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS/MS) was employed for untargeted metabolomics. LC-QTOF-MS/MS identified 63 bioactive compounds across the three endophytes. P. chrysogenum had the highest activity against S. aureus and M. smegmatis (1.15 mg/mL and 0.02 mg/mL, respectively), while P. leucadendri demonstrated moderate activity against M. smegmatis (2.91 mg/mL) and E. coli (1.16 mg/mL). Fusarium sp. exhibited the broadest spectrum of antibacterial activity, with MIC values ranging from 0.03 mg/mL (B. subtilis) to 10 mg/mL (M. smegmatis). P. leucadendri produced 29 metabolites, Fusarium sp. had 23 identified metabolites, and a total of 11 metabolites were identified from P. chrysogenum. The fruits of the plant, accounting for 60%, appeared to be the most abundant in the endophyte diversity when compared to the stems and leaves. This study highlights the potential of fungal endophytes from S. mauritianum as a source of novel bioactive compounds, particularly against multidrug-resistant pathogens, contributing to the ongoing efforts to combat antimicrobial resistance. Full article

Corn straw is considered a renewable biomass energy source, and its unreasonable disposal leads to resource waste and environmental pollution. Black soldier fly (Hermetia illucens L.) larvae (BSFL) facilitate the bioconversion of various types of organic wastes. In this study, we found that 88% of BSFL survived, and 37.4% of corn straw was digested after 14 days of feeding with corn straw. Contrary to expectations, the pretreatment of corn straw with alkaline hydrogen peroxide did not promote its digestion but rather reduced the growth and survival rates of BSFL. Acinetobacter, Dysgonomonas, and unclassified Enterobacteriaceae were the abundant genera in the BSFL gut fed with corn straw. Compared with the standard diet, the relative abundances of carbohydrate metabolism genes, such as the gene abundances of β-glucosidase and α-glucosidase, were higher with corn straw as the substrate. These results suggested that the gut microbial community could regulate suitable and functional microorganisms in response to the substrates. Furthermore, four cellulase-producing strains, namely Klebsiella pneumoniae, Proteus mirabilis, Klebsiella oxytoca, and Providencia rettgeri, were isolated from the guts of corn straw BSFL. These four strains helped increase the conversion rates of corn straw, the weights of BSFL, and survival rates. In summary, we reared BSFL with corn straw and discovered the functions of gut microorganisms in adapting to the substrates. We also isolated four cellulase-producing strains from the BSFL guts and declared the benefits of BSFL digesting corn straw. Full article

The fruit and vegetable industry in post-harvest processing plants is characterized by a substantial consumption of water resources. Wash waters may serve as an environment for the periodic or permanent habitation of microorganisms, particularly if biofilm forms on the inner walls of tanks and flushing channels. Despite the implementation of integrated food safety monitoring systems in numerous countries, foodborne pathogens remain a global public health and food safety concern, particularly for minimally processed food products such as vegetables and fruits. This necessitates the importance of studies that will explore wash water quality to safeguard minimally processed food against foodborne pathogen contamination. Therefore, the current study aimed to isolate and identify bacteria contaminating the wash waters of four fresh-cut processing plants (Poland) and to evaluate the phenotypic antibiotic resistance profiles in selected species. Bacteria were isolated using membrane filtration and identified through mass spectrometry, followed by antibiotic susceptibility testing according to EUCAST guidelines. The results revealed that the level of contamination with total aerobic bacteria in the water ranged from 1.30 × 10⁶ cfu/mL to 2.54 × 10⁸ cfu/mL. Among the isolates, opportunistic pathogens including Enterococcus faecalis, Pseudomonas aeruginosa, Klebsiella oxytoca, Klebsiella pneumoniae, Serratia marcescens, and Proteus vulgaris strains were identified. An especially noteworthy result was the identification of cefepime-resistant K. oxytoca isolates. These findings highlight the importance of monitoring the microbial microflora in minimally processed foods and the need for appropriate sanitary control procedures to minimize the risk of pathogen contamination, ensuring that products remain safe and of high quality throughout the supply chain. Full article

Public health faces daily challenges due to increasing reports of pathogenic microorganisms with new antimicrobial resistance. Klebsiella michiganensis, an emerging pathogen, poses difficulty in its identification using conventional techniques. This study presents the first documented case of NDM-1-producing K. michiganensis in Brazil, identified as the new ST418. Initially, the isolate from a tracheal secretion was misidentified as K. oxytoca. However, accurate identification was achieved through ANI analyses. Whole-genome sequencing was conducted to characterize the genetic context of the resistance genes, to identify virulence factors, and to construct a phylogenetic tree. The bla_NDM-1 gene was found to be harbored on an IncFIB plasmid approximately 112 kb in length, which was transferable in conjugation assays. The detection of carbapenem resistance genes in this species highlights the importance of public health vigilance, as it may serve as a reservoir and disseminator of significant resistance genes. Full article

Spontaneous Bacterial Peritonitis (SBP) is a serious complication and a common cause of death in patients with liver cirrhosis. Between January 2017 and March 2024, a retrospective study was conducted involving 302 patients (>18 years old) with ascites treated at a tertiary referral center in south-eastern Poland. Microbiological analysis of the ascitic fluids was performed in all patients. The presence of microorganisms was found in samples from 17 patients, and 21 pathogens were isolated, including 15 Gram-positive bacteria and 6 Gram-negative bacteria. Staphylococcus epidermidis, MRCNS (methicillin-resistant coagulase-negative staphylococci, resistant to all beta-lactam antibiotics: penicillins, penicillins with beta-lactamase inhibitor, cephalosporins and carbapenems) was the main pathogen detected (19.05%, 4/21), followed by Enterococcus faecalis (9.52%, 2/21), Enterococcus faecium (9.52%, 2/21), Staphylococcus haemolyticus, MRCNS (4.76%, 1/21), Streptococcus mitis (9.52%, 2/21), Streptococcus parasanguinis (9.52%, 2/21), Micrococcus luteus (4.76%, 1/21) and Bacillus spp. (4.76%, 1/21). The following Gram-negative bacteria were also found in the specimens examined: Escherichia coli, ESBL (extended-spectrum β-lactamase producing E. coli) (4.76%, 1/21), Escherichia coli (4.76%, 1/21), Pseudomonas aeruginosa (4.76%, 1/21), Klebsiella oxytoca (9.52%, 2/21) and Sphingomonas paucimobilis (4.76%, 1/21). Gram-positive bacteria caused nosocomial infections in nine patients with SBP, Gram-negative bacteria caused nosocomial infections in two patients. In six patients with SBP, community-acquired infections caused by Gram-negative bacteria were found in three cases, Gram-positive bacteria in two cases, and in one case, community-acquired infection was caused by mixed Gram-positive and Gram-negative. Bacteria isolated from patients with hospital-acquired SBP showed higher drug resistance than those found in patients with non-hospital SBP. Bacterial infections in cirrhotic patients with complications may be responsible for their deteriorating health. Prompt intervention is critical to reducing mortality. Full article

Urinary tract infections occupy a special niche among diseases of infectious etiology. Many microorganisms associated with urinary tract infections, such as Klebsiella oxytoca, Enterococcus spp., Morganella morganii, Moraxella catarrhalis, Pseudomonas aeruginosa, Proteus mirabilis, Staphylococcus aureus, Staphylococcus spp., and Candida spp., can form biofilms. The aim of this research was to study the effect of the enzyme L-lysine-Alpha-oxidase (LO) produced by the fungus Trichoderma harzianum Rifai on the biofilm formation process of microorganisms associated with urinary tract infections. Homogeneous LO showed a more pronounced effect than the culture liquid concentrate (cCL). When adding samples at the beginning of incubation, the maximum inhibition was observed in relation to Enterococcus faecalis 5960—cCL 86%, LO 95%; Enterococcus avium 1669—cCL 85%, LO 94%; Enterococcus cloacae 6392—cCL 83%, LO—98%; and Pseudomonas aeruginosa 3057—cCL 70%, LO—82%. The minimum inhibition was found in Candida spp. Scanning electron microscopy was carried out, and numerous morphological and structural changes were observed in the cells after culturing the bacterial cultures in a medium supplemented with homogeneous LO. For example, abnormal division was detected, manifesting as the appearance of joints in places where the bacteria diverge. Based on the results of this work, we can draw conclusions about the possibility of inhibiting microbial biofilm formation with the use of LO; especially significant inhibition was achieved when the enzyme was added at the beginning of incubation. Thus, LO can be a promising drug candidate for the treatment or prevention of infections associated with biofilm formation. Full article

In the present investigation, the potential of various newly isolated strains which belong to the Enterobacteriaceae family to produce 2,3-butanediol (BDO), an important bio-based compound, was studied. The most interesting strain, namely Klebsiella oxytoca FMCC-197, was selected for further investigation. Commercial (raw) sucrose or molasses, which are important agro-industrial surpluses, were employed as carbon sources for most of the trials performed. Different fermentation parameters (viz. incubation te4mperature, utilization of different carbon sources, substrate inhibition, aeration) were tested to optimize the process. Fermentations under non-aseptic conditions were also conducted to investigate the potential of growth of the strain K. oxytoca FMCC-197 to surpass the growth of other microorganisms in the culture medium and produce BDO. Besides BDO production, in trials in which molasses was employed as the sole carbon source, significant color removal was observed simultaneously with the production of microbial metabolites. The very high BDO concentration ≈115 g L⁻¹ was reported in approximately 64 h during a fed-batch bioreactor experiment, using sucrose and molasses as carbon sources at 30 °C, reaching a conversion yield (Y_BDO) of 0.40 g g⁻¹ and a productivity rate (P_BDO) of 1.80 g L⁻¹ h⁻¹, while similar results were also obtained at 37 °C. The strain demonstrated remarkable results in non-previously sterilized media, as it produced 58.0 g L⁻¹ in 62 h during a fed-batch bioreactor experiment, while the potential to decolorize molasses-based substrates over 40% was also recorded. From the results obtained it is shown that this wild-type strain can be used in large-scale microbial BDO production using various raw materials as fermentative substrates. The wastewater derived after BDO fermentation by K. oxytoca FMCC-197 can be disposed relatively safely into the environment. Full article

The role of bats in the global microbial ecology no doubt is significant due to their unique immune responses, ability to fly, and long lifespan, all contributing to pathogen spread. Some of these animals hibernate during winter, which results in the altering of their physiology. However, gut microbiota shifts during hibernation is little studied. In this research, we studied cultivable gut microbiota composition and diversity of Nyctalus noctula before, during, and after hibernation in a bat rehabilitation center. Gut microorganisms were isolated on a broad spectrum of culture media, counted, and identified with mass spectrometry. Linear modeling was used to investigate associations between microorganism abundance and N. noctula physiological status, and alpha- and beta-diversity indexes were used to explore diversity changes. As a result, most notable changes were observed in Serratia liquefaciens, Hafnia alvei, Staphylococcus sciuri, and Staphylococcus xylosus, which were significantly more highly abundant in hibernating bats, while Citrobacter freundii, Klebsiella oxytoca, Providencia rettgeri, Citrobacter braakii, and Pedicoccus pentosaceus were more abundant in active bats before hibernation. The alpha-diversity was the lowest in hibernating bats, while the beta-diversity differed significantly among all studied periods. Overall, this study shows that hibernation contributes to changes in bat cultivable gut microbiota composition and diversity. Full article

We aimed to determine the prevalence of Demodex spp. and bacterial infection in patients with blepharitis and also to investigate the relationship between culturable microorganisms and Demodex spp. in this study. The study included patients diagnosed with blepharitis (n = 128) and volunteers without ocular problems (n = 113). Eyelash sampling was performed by epilating eight lashes, which were then tested for Demodex spp. using a light microscope. The examination consisted of assessing the patient’s vision with and without ocular correction and tonus in both eyes and a careful examination of the anterior segment of both eyes. Bacterial identification was performed based on morphological, physiological, and biochemical methods. The prevalence of Demodex spp. was 8.0% in patients from the control group and all patients with blepharitis. Isolated forms of Demodex spp. were detected in all infested patients in the control group and in 58% of patients with blepharitis. A total of 35% of patients with blepharitis had from three to nine forms of Demodex spp., and 7% of patients with blepharitis had more than 10 mites in every field of vision. We found a statistically significant relationship between Demodex spp. infestation and the occurrence of eye dryness and sensations of burning and tearing, redness of the conjunctiva, feeling of a foreign body, loss of eyelashes, Meibomian gland dysfunction, and cylindrical dandruff. There were statistically significant relationships between Demodex sp. infestation and the presence of hyperopia, Meibomian cysts, chronic eyelid inflammation, and the use of eyeglasses. There was also a statistically significant relationship between the occurrence of Demodex spp. and seborrheic dermatitis and diabetes mellitus. Culturable microorganisms of the ocular surface were found in 8.7% of participants who were uninfested and in all patients infested with D. folliculorum. We isolated Staphylococcus aureus, Acinetobacter baumannii, Streptococcus pneumoniae, Klebsiella oxytoca, and Bacillus spp. from the conjunctival sac only in patients infested with D. folliculorum. This indicates an increased probability of colonization by pathogenic bacteria in patients with demodicosis. Therefore, patients infested with D. folliculorum should undergo a microbiological examination of conjunctival swabs. Full article

Heavy-metal contaminants are one of the most relevant problems of contemporary agriculture. High toxicity and the ability to accumulate in soils and crops pose a serious threat to food security. To solve this problem, it is necessary to accelerate the pace of restoration of disturbed agricultural lands. Bioremediation is an effective treatment for agricultural soil pollution. It relies on the ability of microorganisms to remove pollutants. The purpose of this study is to create a consortium based on microorganisms isolated from technogenic sites for further development in the field of soil restoration in agriculture. In the study, promising strains that can remove heavy metals from experimental media were selected: Pantoea sp., Achromobacter denitrificans, Klebsiella oxytoca, Rhizobium radiobacter, and Pseudomonas fluorescens. On their basis, consortiums were compiled, which were investigated for the ability to remove heavy metals from nutrient media, as well as to produce phytohormones. The most effective was Consortium D, which included Achromobacter denitrificans, Klebsiella oxytoca, and Rhizobium radiobacter in a ratio of 1:1:2, respectively. The ability of this consortium to produce indole-3-acetic acid and indole-3-butyric acid was 18.03 μg/L and 2.02 μg/L, respectively; the absorption capacity for heavy metals from the experimental media was Cd (56.39 mg/L), Hg (58.03 mg/L), As (61.17 mg/L), Pb (91.13 mg/L), and Ni (98.22 mg/L). Consortium D has also been found to be effective in conditions of mixed heavy-metal contamination. Due to the fact that the further use of the consortium will be focused on the soil of agricultural land cleanup, its ability to intensify the process of phytoremediation has been studied. The combined use of Trifolium pratense L. and the developed consortium ensured the removal of about 32% Pb, 15% As, 13% Hg, 31% Ni, and 25% Cd from the soil. Further research will be aimed at developing a biological product to improve the efficiency of remediation of lands withdrawn from agricultural use. Full article

Gingivitis is accompanied by microorganisms, including pathogens, which must be eliminated to speed up the treatment of inflammation. Laser irradiation may be one of the safe methods for reducing tissue contamination on the tooth surface. The aim of the study was the assessment of the tooth surface in patients with gingivitis after the use of a diode laser to eliminate microorganisms living there. In the first stage of the research, microorganisms were isolated (Candida albicans, C. guilliermondii, Escherichia coli, Haemophilus parainfluenzae, Klebsiella oxytoca, Neisseria subflava, Rothia dentocariosa, Rothia mucilaginosa, Streptococcus pneumoniae) from three patients with gingivitis, their identification confirmed using the MALDI-TOF MS technique (matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry). Then, the irradiation process with a diode laser was optimized to a wavelength of 810 nm ± 10 nm in five variants to reduce microorganisms on the tooth. The tooth surface was analyzed by X-ray photoelectron spectroscopy (XPS) and optical profilometry (OP) before and after irradiation. 103 to 106 CFU were detected on a 0.4 cm² tooth area. Nine types of bacteria and two types of fungi dominated among the microorganisms. The laser at the most effective biocidal dose of 25 W/15.000 Hz/10 µs, average = 3.84 W, with three uses after 15 s, increased the reduction of fungi from 57.97% to 93.80%, and bacteria from 30.67% to 100%. This dose also caused a decrease in the degree of oxidation and in the effect of smoothing on the treated surfaces. Full article

Feruloyl esterase (FAE)-producing micro-organisms to obtain ferulic acid (FA) from natural substrates have good industrial prospects, and the synergistic effect of multiple bacteria can better improve the yield of FA. In this study, on the premise of the synergistic effect of FAE, hemicellulose, and cellulase, the key strain Klebsiella oxytoca Z28 with FAE was combined with CMCase and Xylanase-producing strains to produce FA. The combination of strains with higher FA production are Klebsiella oxytoca Z28, Klebsiella pneumoniae JZE, Bacillus velezensis G1, and their FA production can reach 109.67 μg/g, which is 15% higher than that of single bacteria. To explore the effects of temperature, Ph, inoculum amount, distillers grains concentration and fermentation time on the FAE activity of the combination of strains in the fermentation process, and determined that temperature, Ph, and fermentation time were the main influencing factors and optimized through orthogonal design. The optimized fermentation conditions are 34 °C, Ph 8.0, and fermentation days for 6 days, the FAE activity can reach 270.78 U/L, and the FA yield of the combined strain is 324.50 μg/g, which is 200% higher than that of single-strain fermentation. Full article