Frequency Of Isolation Research Articles

Biological Control of Rapeseed Clubroot (Plasmodiophora brassicae) Using the Endophytic Fungus Didymella macrostoma P2.

Didymella macrostoma P2 was isolated from rapeseed (Brassica napus), and it is an endophyte of rapeseed and an antagonist of three rapeseed pathogens, Botrytis cinerea, Leptosphaeria biglobosa, and Sclerotinia sclerotiorum. However, whether P2 has a suppressive effect on infection of rapeseed by the clubroot pathogen Plasmodiophora brassicae remains unknown. This study was conducted to detect production of antimicrobials by P2 and to determine the efficacy of the antimicrobials and P2 pycnidiospores in suppression of rapeseed clubroot. The results showed that cultural filtrates (CFs) of P2 in potato dextrose broth and the substances in pycnidiospore mucilages exuded from P2 pycnidia were inhibitory to P. brassicae. In the indoor experiment, seeds of the susceptible rapeseed cultivar Zhongshuang No. 9 treated with P2 CF and the P2 pycnidiospore suspension (P2 SS, 1 × 107 spores/ml) reduced clubroot severity by 31 to 70% on the 30-day-old seedlings compared with the control (seeds treated with water). P2 was reisolated from the roots of the seedlings in the treatment of P2 SS; the average isolation frequency in the healthy roots (26%) was much higher than that (5%) in the diseased roots. In the field experiment, seeds of another susceptible rapeseed cultivar, Huayouza 50 (HYZ50), treated with P2 CF, P2 CE (chloroform extract of P2 CF, 30 μg/ml), and P2 SS reduced clubroot severity by 29 to 48% on 60-day-old seedlings and by 28 to 59% on adult plants (220 days old) compared with the control treatment. The three P2 treatments on HYZ50 produced significantly (P < 0.05) higher seed yield than the control treatment on this rapeseed cultivar, and they even generated seed yield similar to that produced by the resistant rapeseed cultivar Shengguang 165R in one of the two seasons. These results suggest that D. macrostoma P2 is an effective biocontrol agent against rapeseed clubroot.

First report of leaf spot disease caused by Cladosporium pseudocladosporioides on Lonicera caerulea L. in Heilongjiang Province, China.

Blue honeysuckle (Lonicera caerulea L.) has contributed to maintaining the forest's ecological balance and remarkable frost-resistant abilities, helping it withstand extremely cold conditions (-46 °C) and a wide pH range (5 to 8) (Sharma and Lee 2021). Between September 2022 and September 2023, leaf spots were observed on approximately 30% of blue honeysuckle plants of the 'Lanjingling' cultivar grown in a 1.13 ha field in Da Hinggan Ling Prefecture (50.32° N, 124.13° E) in Heilongjiang Province, China. The leaves of the affected plants displayed black-colored spots. To identify the causal agents, 10 healthy and symptomatic leaves were randomly collected from ten healthy and infected individual plants, respectively. Small (3 to 4 mm) segments of the symptomatic tissues were immersed in 5% sodium hypochlorite (NaOCl) for 3 min, rinsed three times with sterile distilled water, dried in a paper towel, and plated on 9-cm Petri dishes containing potato dextrose agar (PDA). Ten fungal colonies developed on the PDA plates with an isolation frequency of 100% from 10 symptomatic leaves, and all colonies displayed a morphology consistent with Cladosporium spp. (Bensch et al. 2018). Cladosporium-like fungi were not isolated from healthy leaves. Dark olive-colored mycelia were observed, with straight unbranched conidiophores bearing terminal light brown-colored limoniform conidia (1.80 to 4.50 × 2.10 to 12.60 μm) and surrounded by a thin line of white mycelium (Delisle-Houde et al. 2024). To confirm this identification, PCR amplification of two representative strains LD-299 and LD-300 genomic DNA was performed with ITS1/ITS4 (White et al. 1990) and ACT512F/ACT783R (Carbone and Kohn 1999) primers. Basic local alignment search tool (BLAST) analyses of the National Center for Biotechnology Information database showed that sequences of the ITS (PP600316, PP600317) and ACT (PP624334, PP624335) all revealed 100% (493/493 nt, 493/493 nt; 181/181 nt, 181/181 nt) shared identity with Cladosporium pseudocladosporioides strain ex-type MF473195 and HM148674 (Bensch et al. 2010), respectively. Using a neighbor-joining phylogenetic analysis based on the ITS and ACT sequences, isolates LD-299 and LD-300 clustered in the same clade of C. pseudocladosporioides. Therefore, based on its morphological characteristics and molecular phylogeny, the two isolates were identified as C. pseudocladosporioides (Cosseboom and Hu 2023). A pathogenicity test was performed using nine healthy two-year-old blue honeysuckle Lanjingling plants. Three plants were inoculated with either the LD-299 or the LD-300 conidial suspension (1 × 106 spores/ml) or with clean water as an experimental control (Aydoğdu et al. 2023). All plants were cultured in a greenhouse (28℃, 75% relative humidity, 12 h light and dark cycle), and each experiment was replicated three times. Typical leaf spot symptoms were first observed on the inoculated leaves after 10 days. Morphological and molecular characterization of re-isolated pathogens from the artificially infected leaves indicated that the two isolates were identical, thereby confirming Koch's postulates. Cladosporium pseudocladosporioides previously caused leaf spot disease on artichoke (Cynara scolymus) in Türkiye (Aydoğdu et al. 2023). To the best of our knowledge, this is the first report of C. pseudocladosporioides causing leaf spots on blue honeysuckle in China. Blue honeysuckle production losses due to the leaf spots are critical for growers. Therefore, further focus should be given to investigate the host range and geographic distribution of C. pseudocladosporioides.

Hospitalized COVID-19 Patients with Urinary Tract Infection in Iran: Candida Species Distribution and Antifungal Susceptibility Patterns.

Candida species, typically part of the human skin and mucous membrane flora, can cause opportunistic fungal infections, notably urinary tract infections (UTIs), which are on the rise among hospitalized COVID-19 patients. The lack of understanding of UTIs in this population, coupled with the emergence of multidrug-resistant strains, poses significant challenges for effective treatment and further investigations. In this study, urine samples were collected from 70 COVID-19 patients with UTIs in sterile containers for microbiology examination. After microscopic observation, the isolates were identified both by phenotypic and molecular techniques such as multiplex PCR. Antifungal susceptibility testing (AFST) against fluconazole (Flu), itraconazole (Itr), and amphotericin B (AMB) was performed according to CLSI M27/S4 standard methods, with the frequency of isolates including Candida albicans (n = 20, 51.3%), Candida tropicalis (n = 15, 38.4%), Nakaseomyces glabrata (previously Candida glabrata) (n = 2, 5.1%), Pichia kudriavzevii (previously Candida krusei), and Candida parapsilosis (n = 1, 2.5%). All isolates of C. albicans, C. tropicalis, C. glabrata, and C. parapsilosis were sensitive to amphotericin B, while C. kruzei was resistant to AMB. Around 70% of C. albicans isolates were sensitive to Flu; 20% of C. tropicalis were resistant to itraconazole, while 33% were resistant to fluconazole. C. albicans and C. tropicalis were the main causes of candiduria in infected cases and both Flu and AMB showed good results in AFST in these species. Performing drug susceptibility testing for clinical isolates of Candida spp. provided guidance for appropriate management and control, and timely antifungal treatment.

Frequency and MIC-based Antimicrobial Resistance Profile of MDR and XDR Gram Negative Isolates of Blood Culture Specimens in a Tertiary Care Setting

The escalation of antimicrobial resistance (AMR) among Gram-negative bacteria, particularly multidrug-resistant (MDR) and extensively drug-resistant (XDR) pathogens, posed a significant challenge in tertiary care settings. This paper comprehensively analyzed the antimicrobial resistance profile of MDR and XDR Gram-negative isolates from blood culture specimens at Jinnah Hospital, Lahore, a leading tertiary care facility. Over a period of one year, 9600 blood culture specimens were processed, revealing a high incidence of resistance to a wide array of antibiotics, with notable findings including 100% susceptibility to Colistin and over 90% resistance to several commonly used antibiotics such as Ciprofloxacin and Piperacillin/Tazobactam. Among the positive isolates, 33.8% were classified as MDR and 66.2% as XDR. The study emphasized the critical role of Minimum Inhibitory Concentrations (MICs) in guiding therapeutic decisions, highlighting the wide variability in resistance patterns and the necessity for personalized antimicrobial therapy. The prevalence of MDR and XDR isolates underscored the urgent need for advanced treatment modalities, comprehensive surveillance, and robust antimicrobial stewardship initiatives. Additionally, the study pointed to the emergence of colistin resistance as a critical challenge in managing infections caused by these pathogens. Through detailed analysis and discussion, this study illuminated the grave challenge posed by AMR in tertiary care settings. It emphasized the importance of innovative approaches to antimicrobial stewardship, developing new antimicrobial agents, and ongoing surveillance to effectively combat this public health crisis.

Antimicrobial resistance trends among Klebsiella pneumoniae associated with urinary tract infections in Crete, Greece, 2017-2022.

Klebsiella pneumoniae is one of the most prevalent bacteria causing urinary tract infections (UTIs). Its increasing resistance to a wide array of antibiotics limits available treatment options. This study investigated the characteristics and trends of antimicrobial resistance in K. pneumoniae isolated from UTIs in Crete, Greece, during 2017 and 2022. Among the 11,946 Enterobacteriaceae isolated from urine specimens, a total of 1,771 K. pneumoniae isolates were identified (14.8%), with an isolation frequency secondary to Escherichia coli (66.3%). K. pneumoniae isolates increased over the years, with a peak in the year 2022. Higher resistance rates were detected in ciprofloxacin (41%), trimethoprim/sulfamethoxazole (TMP/SMX) (38.1%) and nitrofurantoin (33.9%). Resistance to ciprofloxacin, amoxicillin/clavulanic acid, tigecycline, and TMP/SMX significantly increased from 33.7%, 24%, 6%, and 33.1%, respectively, over the years 2017-2019, to 47.8%, 34.2%, 14.3% and 42.8%, respectively, over the period 2020-2022. ESBL production and carbapenem resistance were decreased by 2.2% and 3.7%, respectively, over the two three-year periods (2017-2019 and 2020-2022). Among the 278 carbapenem-resistant K.pneumoniae (CRKP) isolates, 164 (59%), 66 (23.7%), 18 (6.5%) and 16 (5.8%) were positive for KPC, NDM, VIM and OXA-48 enzymes, respectively. Only 14 (5%) isolates harboured two carbapenemase genes, namely 10 (3.6%) both blaNDM and blaVIM, and 4 (1.4%) both blaKPC and blaNDM. Females, inpatients and the elderly were more frequently affected by CRKP. The frequency of multidrug-resistant (MDR) and extensively drug-resistant (XDR) isolates were 32.6% and 7.7%, respectively. Continuous surveillance of local microbial prevalence and monitoring of antimicrobial resistance patterns provide critical information to guide the empiric therapy for UTIs and control the spread of MDR bacteria.

Multi-year Epidemiology of Nontuberculous Mycobacteria Across a Diverse Healthcare System

Background: Nontuberculous mycobacteria (NTM) are ubiquitous potential pathogens implicated in healthcare-associated outbreaks. There is a paucity of studies describing transmission risk in the healthcare setting. To estimate the potential healthcare-associated transmission of NTM, we characterized the frequency of NTM clinical isolates across our multi-facility healthcare system. Method: We performed a retrospective review of all clinical NTM isolates at 21 healthcare facilities in a large health system between January 2019 through June 2023 (inclusion criteria: all first unique species). We analyzed the quarterly frequency of isolates for each species, by facility. We identified higher-than-expected species frequencies, which was defined as a quarterly frequency ≥50% higher than the average quarterly frequency for that facility, for the entire study period (analysis omitted for any hospital with an average quarterly frequency 10 unique patient isolates in any 12-month period or &gt;2 in a single month except for M. abscessus at Hospital A. The quarterly frequency of the three most common species among hospitals with ≥2 unique isolates per 12-month period are displayed in figure 2. An increase of 50% from the average quarterly infection count occurred 34 times for M. avium complex across 10 hospitals, 14 times for M. gordonae across 2 hospitals and 14 times for M. lentiflavum across 2 hospitals (Figure 3). Conclusion: A diverse group of NTM were isolated across our healthcare system over the study period, most commonly M. avium complex, M. gordonae, and M. lentiflavum, each with hospital-specific temporal frequencies that suggest the potential for undetected outbreaks, while frequencies of less commonly isolated species were rarely suggestive of potential undetected outbreaks. Further epidemiologic investigation of in-hospital transmission routes, with whole genome sequencing to determine genetic relatedness, is necessary to identify undetected outbreaks.

Comparison of Real-time Polymerase Chain Reaction and Culture for Targeting Pathogens in Pediatric Severe Community-Acquired Pneumonia.

This study aims to determine the frequency of pathogen detection by real-time polymerase chain reaction (PCR), the frequency of pathogen isolation by culture; and compare the value of real-time PCR and culture of nasopharyngeal aspiration samples in patients with severe community-acquired pneumonia (sCAP). It was a prospective and descriptive study. All pediatric patients diagnosed with sCAP were performed real-time PCR and culture of nasopharyngeal aspiration samples. A total of 336 patient samples were obtained from children with sCAP. Real-time PCR detected pathogens in 312 patients (92.9%), while culture isolated bacteria in 228 patients (67.9%). Coinfections were reported in 279 cases (83.0%) through real-time PCR. The frequency of agreement between culture and real-time PCR was quite high (P < .001). Real-time PCR demonstrated more ability for detecting microorganisms than culture. This finding highlighted the value of real-time PCR for targeting pathogens in children with sCAP, particularly in cases involving complex pathogens or those requiring timely identification.

Impact of MIC Breakpoint Changes for Enterobacterales on Trends of Antibiotic Susceptibilities in An Academic Medical Center

Background: The Clinical &amp; Laboratory Standards Institute (CLSI) recommends use of annual antibiograms to help guide empiric antibiotic therapy. Because CLSI periodically updates minimum inhibitory concentration (MIC) breakpoints, we assessed the impact of these updates on longitudinal trends in antibiotic susceptibility rates for Escherichia coli and Klebsiella pneumoniae at a single academic medical center in Atlanta, GA. Methods: Susceptibilities for cefepime, ceftazidime, and levofloxacin in E. coli and K. pneumoniae were extracted from hospital antibiograms from 1988 to 2022. Starting in 1995, intensive care units (ICUs) and wards had separate annual antibiograms, which we combined using weighted averages to create annual overall hospital antibiograms. After summarizing the frequency of isolates tested and susceptibilities using medians and interquartile ranges (IQR), we conducted an interrupted time series analysis using linear segmented regression models, to evaluate the level changes and trends in susceptibility, before and after CLSI MIC breakpoints were updated for ceftazidime (2010 and 2017), cefepime (2014 and 2017), and levofloxacin (2013). Results: Among 21,214 E. coli, there was a median of 291 [IQR: 104, 555] isolates tested annually. Similarly, among 8,686 K. pneumoniae isolates, the median was 125 per year (IQR: 76, 178). Prior to the MIC breakpoint changes, baseline susceptibility trends of both organisms to all 3 antibiotics significantly declined at a rate between 0.2% to 2.4% per year (Table 1). For cefepime (Figure 1), susceptibility decreased annually during 1988 – 2013 for both E. coli (-0.5%) and K. pneumoniae (-1.2%). There were no significant level changes but there were trend changes after 2018, for E. coli (+2.1%) and K. pneumoniae (– 5.5%). For ceftazidime (Figure 2), significant level changes occurred after 2010 for both organisms (E. coli: -5.7%; K. pneumoniae: -5.2%). For levofloxacin (Figure 3), the breakpoint update in 2013 lead to significant level change in susceptibility (E. coli: +8.4%; K. pneumoniae: +11.4%). Conclusion: Overall, we observed a consistent decrease in antibiotic susceptibility in E. coli and K. pneumoniae over three decades, with immediate increases in the level change of susceptibility when MIC breakpoints were changed, followed by a decreasing trend. These findings highlight the importance of longitudinal surveillance and MIC breakpoint changes to inform antimicrobial stewardship strategies.

Modeling and analysis of a two-stage tri-linear pendulum ultra-low frequency horizontal vibration isolator with experimental investigations.

In engineering practice, the external horizontal oscillations always influence the working performance of precise instruments, advanced manufacture equipment, and gravitational wave detection. In order to ensure the normal operation of these instruments, it is necessary to attenuate these vibrations adequately. The pendulum mechanism horizontal vibration isolator is an efficient method. Hence, this paper presents a type of two-stage tri-linear pendulum horizontal vibration isolator (TPHVI). The first-stage TPHVI is connected in series with the second-stage one. The dynamic equations of the two-stage TPHVI are subsequently established so that the vibration isolation performance of the two-stage TPHVI is acquired. The analysis result of the natural frequency of the two-stage TPHVI reveals that it can obtain a low frequency vibration isolation performance when the first-stage TPHVI swings in a small value. As a case study, an experimental rig is constructed. The measured transmissibility keeps in good agreement with the calculated one. The natural frequency of the second-stage TPHVI is 0.25Hz. The initial vibration isolation frequency is 0.3Hz. When the external frequency is 0.8Hz, the transmissibility of the second-stage TPHVI reaches -20 dB. Meanwhile, when the external frequency is 3Hz, the transmissibility of the second-stage TPHVI is -40 dB. These measured data demonstrate that the proposed two-stage TPHVI can realize low frequency vibration isolation horizontally, which will have broad application prospects in the field of ultra-precision in engineering practice.

Study of microbiological activity of topical antibacterial agents for treatment of combat injuries

The problems of combat injuries are determined by the issue of treatment of purulent complications, because wounds received during combat operations are usually infected, and the most serious complication of such wounds is the development of purulent infection. Objective — to study the features of the aerobic component of combat wounds, the sensitivity of causative agents of wound infection to antibacterial drugs, and the antimicrobial activity of individual drugs in relation to extensively resistant and polyresistant microbial agents in order to develop external therapy algorithms as part of the complex treatment of these injuries. Materials and methods. The data of microbiological studies of patients with combat wounds were used in the work. Results and discussion. Based on the results of monitoring causative agents of purulent­inflammatory complications of wound injuries, a range of leading types of microorganisms was revealed. The isolation of flora resistant to most antibiotics from wounds dictates the need to change the strategy of infection control with strengthening measures to prevent nosocomial transmission, which should be considered when prescribing antibiotic therapy. Conclusions. Based on the results of monitoring the causative agents of purulent-inflammatory complications of wound injuries, a range of leading types of microorganisms was revealed: K. pneumoniae, E. faecalis, S. haemolyticus, P. aeruginosa, E. coli. The dominant microorganisms (by isolation frequency) in the wound contents were: staphylococci (3.0—5.69 lg CFU/ml), enterobacteria (4.69—7.0 lg CFU/ml), enterococci (3.0—4.69 lg CFU/ml). In addition, aerobic bacilli (3.0—3.69 lg CFU/g) and pseudomonads (5.69—7.0 lg CFU/g) were determined in the wound contents, which did not have a dominant character.The use of a topical agent containing dioxidin can contribute to the improvement of the effectiveness of the local treatment of purulent wounds infected with strains of gram-negative bacteria, in particular E. coli, K. pneumoniae and P. aeruginosa, with poly and extensive resistance to antibiotics both in monocultures and in microbial associations, in particular with strains of S. aureus, including MRSA.

Surveying the Distribution and Diversity of Predominant Mycobiota in Cultivated and Uncultivated Soils of Zaria, Northwestern Nigeria

Study’s Novelty/Excerpt This study presents a comprehensive survey of the distribution and diversity of soil fungi in cultivated and uncultivated soils of Zaria, Northwestern Nigeria, highlighting their morphological and microscopic characteristics, frequency, and relationship with physicochemical properties. It identifies and quantifies the presence of eight different fungal species, with Aspergillus niger being the most dominant, providing critical insights into the fungal biodiversity of this region. Additionally, the study correlates the richness of soil mycobiota with specific soil properties such as pH, organic matter content, and moisture levels, suggesting that these factors significantly influence fungal diversity and distribution, and recommending further research to assess their impact on soil productivity. Full Abstract Soil fungi are a crucial component of terrestrial ecosystems, playing a vital role as decomposers in nutrient cycles. The diversity of soil fungi serves as an indicator of soil health (favorable/deficient). Understanding soil biodiversity and its biological functioning will help not only in crop yields but also in a healthier society as it directly or indirectly enters the food chain. This study aimed at surveying the distribution and diversity of predominant mycobiota in cultivated and uncultivated soils of Zaria, Northwestern Nigeria. Focusing on their morphological and microscopic characteristics, percentage frequency, and relationship with physicochemical properties. After removing the surface soil, soil samples were randomly collected from two locations at 0-15 cm depths. Fungi were isolated via the dilution method on PDA. Previously established criteria were used to identify eight different fungal species: Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Fusarium spp., Mucor spp., Penicillium spp., Rhizopus spp., and Trichoderma spp. Aspergillus niger was the most dominant species, with a relative frequency of 26%, followed by Fusarium spp. and Rhizopus spp. With 18% and 17%, respectively. Aspergillus fumigatus was the least abundant (2%). The highest fungal isolation frequency (32%) was observed in L1 cultivated soil, closely followed by L2 uncultivated soil (31%), with the lowest in L2 uncultivated soil (16%). Physicochemical analysis indicated that these soils are rich in mycobiota, attributed to a basic pH range of 7.12-9.15, high organic matter content, and optimal moisture levels. Soils with significant organic matter due to litter accumulation, moist deciduous vegetation, and neutral to alkaline pH levels retain considerable moisture content, fostering a rich qualitative and quantitative mycobiota for recycling dead organic matter making nutrients available for suitable utilization. Further research is recommended to evaluate the impact of these isolated fungi on soil productivity, to determine whether their presence has a positive or negative effect.

Multi-degree-of-freedom ultra-low frequency vibration isolation system for precision devices with decoupled control

High precision devices are sensitive to low frequency background vibration. In order to meet the requirement of vibration isolation and adaptation to different working modes, a multi-DOF (Degree-of-Freedom) ultra-low frequency vibration isolation system is proposed. The system uses diaphragm bellow isolators to bear static load and electromagnetic actuators to exert control forces. Auxiliary isolators of variable stiffness are used to make the system adaptive to operation and transportation. The dynamic model of the vibration isolation system is established to analyze vibration characteristics and investigate control laws. A broadband control law based on the actuation-measurement state decoupling is proposed and the mixed feedback and feedforward control is adopted to realize ultra-low frequency vibration isolation. Numerical and experimental results have demonstrated that the proposed system is able to isolate ground vibration effectively. The initial vibration isolation frequency is reduced to 0.5 Hz and the transmissibility reaches −8.3 dB at 1 Hz and −21.9 dB at 2 Hz.

Phenotypic and genotypic characteristics of mecA - positive oxacillin-sensitive Staphylococcus aureus isolated from patients with bloodstream infection in a tertiary hospital in Southern Brazil.

Staphylococcus aureus are extremely important microorganisms, either from an epidemiological point of view or as a pathogen, responsible for causing a series of infectious processes, whether simple, restricted to the skin, or invasive infections such as bacteremia. The emergence of Oxacillin Sensitive-Methicillin Resistant S.aureus (OS-MRSA) isolates has imposed difficulties in the treatment of patients with staphylococcal infection, as such isolates can be mistakenly classified as sensitive and lead to failure of the therapy used. Thus, the objective of this study is to evaluate the prevalence, and genotypic and phenotypic characteristics, of OS-MRSA isolates, from bloodstream infections, collected from patients admitted to a hospital in southern Brazil, as well as to evaluate the treatment used. For this, 801 unique isolates of S. aureus, collected from blood cultures, between January 2011 and December 2020 were evaluated. Of these, 96 isolates were classified as sensitive to oxacillin. The isolates were identified and had their sensitivity profile performed by manual and automated methods. The minimum inhibitory concentration for vancomycin, daptomycin, oxacillin, linezolid and teicoplanine was performed by e-test. The mecA, vanA genes, typing of the SCCmec elements, as well as the search for the icaA, tst-1 and pvl virulence genes were performed by PCR. Biofilm formation was performed using the crystal violet technique. The Sequence Type (ST), as well as the Clonal Complex (CC) of the isolates was evaluated by the RTq -PCR. The clinical characteristics of the patients were evaluated through an active search in medical records. After investigating the mecA gene, 27.1% (26/96) of the isolates were considered OS-MRSA, with SCCmec type I being the most prevalent, 46.1% (12/26). Among the evaluated isolates, 41% (9/22) were included in CC5 and ST9. As for virulence, all isolates were positive for the icaA gene and characterized as strong biofilm formers. The pvl gene was found in 92.3% (24/26) of the isolates and the toxic shock syndrome toxin was present in 61.5% of the isolates (16/26). All isolates were negative for the presence of the van A gene. As for the clinical outcome, 73% (19/26) of the patients were discharged from the hospital and 27% (7/26) died. It was possible to observe a high frequency of OS-MRSA isolates causing bloodstream infections. Furthermore, such isolates contain several virulence genes, which may contribute to a worse clinical outcome.

MICROFLORA OF COMBAT WOUNDS OF THE MALE PELVIC ORGANS AND DYSBACTERIOSIS OF THE URINARY SYSTEM

Introduction. Identification of microorganisms that colonize combat wounds and cause wound infection is of primary importance for the subsequent successful treatment of the patient. The resistance of microorganisms to antimicrobial drugs makes the efforts of modern medicine in the fight against infectious agents ineffective. The problem of infertility is closely related to combat injuries, their infection, stress, and neurotic disorders. Aim. Obtaining and summarizing data on microbial colonization of mine-blast wounds of pelvic organs and the microbiome of the genitourinary system of combatants. Materials and methods. 84 smears were taken from 56 wounds of 36 patients with injuries of the pelvic organs who were being treated. 73 patients with injuries were examined for the presence of mycoflora in the urogenital tract. Isolation of pure bacterial cultures was carried out by inoculating the studied material using meat-peptone agar, blood agar, chromogenic agars. For the diagnosis of urogenital or other infections by the PCR method, a scraping from the back wall of the urethra was taken from the patients. Results. Predominant microorganisms in positive cultures of smears were non-fermenting gram-negative rods, which in 28% of cultures belonged to the genus Acinetobacter, in 26% to the genus Pseudomonas. As for associated infections, 20% of them consisted of the genus Acinetobacter, 32% – Enterobacter, 4% – Klebsiella and 29% – Pseudomonas. Gram-positive cocci were isolated in 37% of positive smear cultures. The frequency of isolation of the genus Streptococcus in monoinfection was 2.5%, followed by the genus Clostridium – 2%, Bacillus – 3%, Enterococcus – 4% and Actynomycceas – 4%. In associated infections, the frequency of isolation of the genus Streptococcus was 4%, followed by the genus Clostridium – 2%, Bacillus – 4%, Enterococcus – 3% and Actynomycceas – 5%. When analyzing the microflora of the genitourinary system, it was found that the priority role belongs to the combined infection, when there are associations of specific pathogens such as Ureaplasma spp., Mycoplasma spp., Chlamidia spp., Neisseria gonorrhoeae, Trichomonas vaginalis, Streptococcus spp., Enterococcus faecalis, which is 80% of the entire microbiome. Conclusions. Acinetobacter baumanii and Klebsiella pneumoniae are the dominant microflora complicating the course of combat wounds during almost two years of Russia's full-scale war against Ukraine. Probably, the duration of hostilities, the large number of wounded, and the forced mass unsystematic use of various antibiotics lead to rapid changes in the spectrum of pathogens of combat wounds. Combat wounds and their infection, stress, and nervous disorders lead to an imbalance of microflora, in particular microflora of the genitourinary system, which can be one of the causes of infertility. Chlamydia and Ureaplasma are the most common microorganisms that colonize the urogenital tract of men injured as a result of hostilities.

Isolation and morphological identification of fungi from endometrial samples of postpartum dairy cows

Numerous studies on endometritis of postpartum dairy cows describe the bacterial aetiology of the condition. However, studies on fungal agents causing inflammatory conditions of the reproductive tract are limited. The objective of the present study was to identify the mycotic isolates from the reproductive tract of postpartum dairy cows and analyse their susceptibility to antifungal agents. In this study, 70 endometrial cytobrush samples were collected from dairy cows 40 - 120 days postpartum (dpp) and cultured in Sabouraud dextrose agar (SDA). The isolated fungal cultures were identified by Lactophenol Cotton Blue staining and were subjected to antifungal susceptibility testing. The frequency of isolation of fungi from endometrial samples was 15.71 per cent (11/70). Aspergillus niger was isolated from four (33.33%) samples, followed by Penicillium spp. (25%), Aspergillus fumigatus (16.67%), Mucor spp. (16.67%) and Absidia spp. (8.33%). Antifungal susceptibility testing revealed that 50 per cent of the isolates were susceptible to fluconazole, whereas only 25 per cent of the isolates were completely resistant to amphotericin-B. The prevalence of fungal infection in reproductive tract was higher in cows with an age of ≥ 5 years and a parity of ≥ 2, though this was not significant. A significantly higher number of cattle with history of intrauterine antibiotic therapy were positive for fungal culture. However, the association between prevalence of fungi and reproductive complications was statistically non-significant. Keywords: Postpartum dairy cows, fungal infections, Aspergillus niger, mycotic infections

First Report of Anthracnose on Banana Peppers Caused By Colletotrichum scovillei in New York.

In early August 2023, a disease outbreak on hot banana peppers (Capsicum annuum cv. Golden Dagger) was reported in Cattaraugus County, New York (NY). Disease incidence was at least 60%. Affected developing and mature fruit had at least one tan, soft, sunken lesion with salmon-colored spore masses surrounded by brown, necrotic margins. Microscopic observation of the lesions identified acervuli and setae typical of Colletotrichum spp. Isolations were made from these lesions by spreading conidia from the acervuli on 2% water agar (WA) + 0.02% (w/v) ampicillin. Colonies were hyphal tipped and transferred onto clarified V8 juice agar (CV8) and incubated at 20°C. The isolation frequency was 100% and a total of six isolates were obtained: Coll23Pep001, Coll23Pep003, Coll23Pep005, Coll23Pep007, Coll23Pep008, and Coll23Pep010. After 10 days, colonies were subcultured to potato dextrose agar (PDA) and CV8. On PDA, colonies appeared off-white to dark gray with sparse aerial mycelia. On CV8, the colony was pale gray with acervuli and orange-colored spore masses in the center. Conidia were hyaline, smooth and fusiform to round, and tapered at both ends. Mean conidial dimensions (n = 20) were 20.2 (13.75 to 25) µm long × 4.7 (3 to 6.25) µm wide. To confirm the identity of the isolates, DNA was extracted, and PCR performed to amplify the internal transcriber spacer (ITS) region (primers ITS1/ITS4; White et al. 1990), and actin (ACT) (primers ACT-512F/ACT-783R; Carbone and Kohn 1999) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes (primers GDF1/GDR1; Guerber et al. 2003). Pairwise alignment of the sequences showed all isolates had 100% similarity to C. scovillei ex. holotype CBS 126529 (Damm et al. 2012). Sequences from all isolates were deposited in GenBank with accessions PP556967 to PP556972 (ITS), PP565766 to PP565771 (ACT), and PP565772 to PP565777 (GAPDH). For pathogenicity testing, all isolates were grown on CV8 at 20°C in the dark for 10 days. Conidia were harvested by flooding the plates of each isolate with sterile distilled water and filtering the suspension through a double layer of cheesecloth. The concentration of the conidial suspension was adjusted to 5 × 105 spores per ml. Pathogenicity of the six isolates was tested on banana pepper fruit by using a sterile toothpick to pierce the skin at the two opposite ends. A droplet (10 µl) of the conidial suspension was placed on each wound. The same number of fruit were inoculated without wounding, and non-inoculated control fruit received a droplet of sterile distilled water (either wounded or unwounded). The experiment was repeated twice. All fruit were placed in a humid box at room temperature for 7 days. All wounded and inoculated fruit developed sunken lesions filled with salmon-colored conidial masses. Disease did not occur on the unwounded, inoculated fruit nor the non-inoculated controls. C. scovillei was recovered from all inoculated fruit by reisolating onto CV8 media and isolates had similar morphology and conidial dimensions to the original isolates. To the best of our knowledge, this is the first report of C. scovillei causing anthracnose on pepper in NY. C. scovillei has been reported in South Carolina (Toporek and Keinath 2021), Brazil (Caires et al. 2014), eastern Asia (de Silva et al. 2019), and Kosovo (Xhemali et al. 2023). The pathogen is particularly aggressive on pepper and poses substantial threats to pepper production around the world.

Risk of fungal exposure in the homes of patients with hematologic malignancies

BackgroundPatients with hematological malignancies are at a high risk of developing invasive fungal infections (IFI) because they undergo several cycles of treatment leading to episodes of neutropenia. In addition, they alternate between hospital stays and periods spent at home. Thus, when an IFI is diagnosed during their hospital stays, it is highly challenging to identify the origin of the fungal contamination. The objective of this study was to analyze at home fungal exposure of 20 patients with leukemia by taking air and water samples in their living residence. MethodsAir was sampled in 3 rooms of each home with a portable air system impactor. Tap water was collected at 3 water distribution points of each home. For positive samples, fungi were identified by mass spectrometry or on the basis of their morphological features. Results85 % of homes revealed the presence in air of Aspergillus spp. and those belonging to the section Fumigati presented the highest concentrations and the greatest frequency of isolation. Concerning mucorales, Rhizopus spp. and Mucor spp. were isolated in air of 20 % and 5 % of dwellings, respectively. In 4 homes, more than 70 % of the fungal species identified in air were potential opportunists; these were mainly Aspergillus spp. with concentrations greater than 20 cfu/m3. The water samples revealed the presence of Fusarium in 3 dwellings, with concentrations up to 80 cfu/L. Finally, for one patient, fungal species isolated during a period of hospitalization were phenotypically similar to those isolated in samples taken at home. For a second patient, a PCR Mucorale was positive on a sample of bronchoalveolar fluid while air samples taken at his home also revealed also the presence of mucorales. ConclusionThe presence of opportunistic fungal species in the air of all the explored homes suggests the need for strengthened preventive measures in the home of immunocompromised patients. It would be interesting to compare the fungi isolated (from patients and from their environment) by genotyping studies aimed at specifying the correspondence existing between fungal species present in the patients’ homes and those responsible for IFI in the same patients.

The sweating process promotes toxigenic fungi expansion and increases the risk of combined contamination of mycotoxins in Radix Dipsaci.

Sweating is one of the most important processing methods of Chinese medicinal herbs. However, the high temperature and humidity environment required for sweating Chinese medicinal herbs makes it very easy for fungi to breed, especially toxigenic fungi. The mycotoxins produced by these fungi will then contaminate the Chinese medicinal herbs. In this study, we explored the changes in mycobiota, toxigenic fungi, and mycotoxins with and without sweating in Radix Dipsaci (RD), a typical representative of traditional Chinese medicine that requires processing through sweating. We also isolated and identified the toxigenic fungi from RD, whether they were subjected to sweating treatment or not, and examined their toxigenic genes and ability. The results showed that the detection rate of mycotoxins (aflatoxins, ochratoxins, zearalenone, and T-2 toxin) in RD with sweating was 36%, which was 2.25-fold higher than that in RD without sweating. We also detected T-2 toxin in the RD with sweating, whereas it was not found in the RD without sweating. The sweating process altered the fungal composition and increased the abundance of Fusarium and Aspergillus in RD. Aspergillus and Fusarium were the most frequently contaminating fungi in the RD. Morphological and molecular identification confirmed the presence of key toxigenic fungal strains in RD samples, including A. flavus, A. westerdijkiae, F. oxysporum and F. graminearum. These four fungi, respectively, carried AflR, PKS, Tri7, and PKS14, which were key genes for the biosynthesis of aflatoxins, ochratoxins, zearalenone, and T-2 toxin. The toxigenic ability of these four fungal strains was verified in different matrices. We also found that A. flavus, A. westerdijkiae, and F. oxysporum were isolated in RD both with sweating and without sweating, but their isolation frequency was significantly higher in the RD with sweating than in the RD without sweating. F. graminearum was not isolated from RD without sweating, but it was isolated from RD with sweating. These findings suggest that the sweating process promotes the expansion of toxigenic fungi and increases the risk of combined mycotoxin contamination in RD.

Infection route associated with invasive group A streptococcal toxic shock syndrome in maternal deaths: Nationwide analysis of maternal mortalities in Japan

ObjectivesTo clarify the infection route in maternal death due to invasive group A streptococcal (GAS) infection and toxic shock syndrome (TSS). MethodsA retrospective study was conducted on maternal deaths due to GAS-TSS in Japan between January 2010 and March 2024. The final causal diagnosis of maternal death and the infection routes of GAS were analysed using medical records, laboratory data and autopsy findings. ResultsAmong the 616 maternal deaths during the study period, 48 (8%) involved infectious diseases. The most common infection was invasive GAS (56%, n = 27), 21 (78%) and six cases occurred during the antepartum and puerperium periods, respectively. In the GAS-TSS group, 71% (15/21) infections were originated the upper respiratory tract. However, in the puerperium cases, 67% (4/6) were infected from the genital tract. In addition, no maternal deaths due to GAS-TSS were reported during the COVID-19 pandemic period in Japan from 2020 to 2023. ConclusionMost antepartum GAS infections were from the upper respiratory tract. They may be reduced by preventive measures, including frequent disinfection, wearing masks and isolation from persons at high risk of carrying GAS, such as symptomatic children. On the other hand, GAS-TSS during puerperium infection via the genital tract.

Bacterial Colonization on Endoscopic Materials and Surgical Field Without Infections After Transoral Endoscopic Thyroidectomy.

Our aim was to determine whether bacteria contamination occurred within the surgical field or on endoscopic equipment during surgery using the transoral endoscopic thyroidectomy vestibular approach (TOETVA). Participants were recruited from patients planned for TOETVA between May 2017 and December 2019. Bacterial samples were taken before and at the conclusion of the TOETVA procedure. The preoperative and postoperative samples were taken from the endoscopic materials and inferior oral vestibulum using a sterile flocked swab. The study resulted in 480 samples (80 TOETVAs). No vestibular, port site, or neck infections occurred in any of the patients. Three (3.7%) out of 80 patients developed postoperative fever. Our results show different microbial communities during TOETVA. The most prevalent species detected were S treptococcus species. Multivariate logistic regression analyses revealed that the degree of contamination depended on the sampling site (inferior vestibulum > equipment) ( P =0.03). In addition, the abundance of bacteria was affected by operative time ( P =0.013). There were no significant differences observed in isolation frequencies of bacteria in malignancy ( P =0.34). TOETVA surgery is categorized as a "clean-contaminated" operation. A swab identified the common colonizers of oral microbiota on the endoscopic equipment and within the surgical field.