Bacterial Culture Research Articles

Klebsiella pneumoniae Complex-Associated Peritonitis, Lymphadenitis, and Pyelonephritis in Juvenile Raccoons (Procyon lotor) under Rehabilitator Care in New York, New Jersey, and Wisconsin, USA.

Klebsiella spp. are gram-negative facultative anaerobic heavily encapsulated bacteria associated with opportunistic and primary infections in a wide range of species. We assessed a series of cases (n=8) of necrosuppurative peritonitis, lymphadenitis, and/or pyelonephritis in wild juvenile raccoons (Procyon lotor) that died under rehabilitator care in New York, New Jersey, and Wisconsin, US, between July 2020 and December 2023, plus a retrospective case of a juvenile raccoon necropsied from New York in August 2011. Gross necropsy (n=9) and histopathology (n=9) were performed to characterize the lesions, whereas bacterial culture (n=8) was used to identify and characterize the bacteria and associated phenotype. We observed gram-negative short rods and coccobacilli (7/9; 78%), fibrinosuppurative peritonitis of variable severity (7/9; 78%) correlated to gross pyoabdomen (5/9; 56%) or abscessation (2/9; 22%), lymphadenomegaly and associated necrosuppurative lymphadenitis (5/9; 56%), and urinary tract disease (3/9; 33%). Aerobic culture of affected tissues isolated Klebsiella pneumoniae (n=4), K. pneumoniae subsp. ozaenae (n=2), Klebsiella variicola (n=1), and Klebsiella sp. (n=1). Our study strongly suggests an association of bacteria within the K. pneumoniae complex with peritonitis, lymphadenitis, and pyelonephritis in raccoons. Disease might be associated with underlying nosocomial infection given that all animals were under rehabilitator care at the time of death.

Zero incidence of culture-positive septic arthritis and low infection rate following ACLR with all-soft tissue quadriceps tendon autograft: An analysis of 1053 cases.

To evaluate the infection rate following anterior cruciate ligament reconstruction (ACLR) using all-soft tissue quadriceps tendon (ASTQT) autograft. All primary ASTQT autograft ACLRs within a single surgeon's prospectively collected database from 2011 to 2021 were retrospectively reviewed. No topical antibiotics were administered and no graft-soaking with antibiotics was performed during the study period. Patients who underwent multiligament knee reconstruction or a cartilage restoration procedure were excluded. Patients who underwent a subsequent procedure, including irrigation and debridement (I&D) of the knee joint, were included. Case-specific data, including fluid culture analysis, antibiotics (type, route of administration and duration), time to debridement and method of debridement, were collected. Descriptive statistics were utilized to analyze demographics, incidence and possible association between the need for I&D and concomitant meniscus surgery. Out of 1053 cases (mean age: 20.2 ± 6.3, 44.6% female), four patients (0.38%) (mean age: 18.5 ± 4.0, 25% female) underwent subsequent I&D (arthroscopic I&D only [n = 1], graft harvest site I&D only [n = 1], combined graft harvest site and arthroscopic I&D [n = 1] and tibia wound and arthroscopic I&D [n = 1]). There was no significant difference with regard to demographics. No joint fluid cultures returned positive; one wound culture returned positive for Methicillin-susceptible Staphylococcus aureus from their graft harvest site. The time to I&D ranged from 18 to 23 days. Concomitant meniscectomy or meniscal repair was not associated with requiring surgical I&D. All patients who underwent I&D were prescribed antibiotics for a minimum of 10 days and a maximum of 31 days (mean: 16.25 days). Three patients (75%) who underwent I&D ultimately returned to sport. One patient was lost to follow-up. ACL grafts were retained in all patients. The incidence of culture-positive septic arthritis following ASTQT autograft ACLR is 0%, while the overall need for I&D of 0.38% is low and not related to concomitant meniscal procedures or patient-specific factors. Level IV.

Black raspberry modulates cecal and oral microbiome at the early stage of a dibenzo[def,p]chrysene-induced murine oral cancer model.

While tobacco smoking is a risk factor in the development of oral squamous cell carcinoma (OSCC), only a fraction of smokers develop the disease. Compelling evidence shows that microbial community composition is associated with carcinogenesis, suggesting that the microbiome may play a role in cancer development of smokers. We previously showed that black raspberry (BRB) protects against OSCC induced by the tobacco constituent dibenzo[def,p]chrysene (DBP) altering genetic and epigenetic markers in a manner consistent with its cancer preventive activity. In the present study, we conducted a mouse experiment to investigate the effects of BRB and DBP individually and in combination on the oral and gut microbiota. DBP-induced DNA damage in the mouse oral cavity which is an essential step for the development of OSCC in mice. 16S rRNA gene sequencing revealed that BRB significantly increased microbial diversity and shifted microbiome composition in the gut and oral cavity, whereas DBP had no significant effect. In both gut and oral microbiota, Akkermansia muciniphila was significantly reduced after BRB treatment; however, this was not consistent with pure culture in vitro assays suggesting that the impact of BRB on A. muciniphila may be mediated through indirect mechanisms including the host or other microbes. Indeed BRB, but not DBP, was found to modulate the growth kinetics of human gut microbes in vitro including lactic acid bacteria and Bacteroides spp. The results of the current study further emphasize the interplay of microbiome and environmental factors in the development and prevention of OSCC.

A New Approach to Evaluate the Bactericidal Activity of Different Antiseptic Ophthalmic Preparations Used as Surgical Prophylaxis

Background: A survey conducted by the European Observatory on Cataract Surgery showed high heterogeneity in the use of antiseptics both preoperatively and in the operating room, highlighting the absence of a global consensus regarding ocular infection prophylaxis in cataract surgery. Methods: The antibacterial activity of seven antiseptic ophthalmic formulations (AOFs) registered as medical devices and the two most common disinfectants were evaluated in vitro against five bacterial species. The viability of bacterial strains after exposure to the antiseptic was evaluated with different techniques: the in vitro Minimum Inhibitory Concentration and the subsequent Minimum Bactericidal Concentration, performed on liquid and solid culture medium. Furthermore, a real-time assessment of bacterial viability was conducted using double staining for live/dead bacterial cells by fluorimetric assay. This evaluation was performed on both the time-killing curve and the tear dilution effect test. Results: We observed a high variability across the different AOFs in terms of inhibitory/bactericidal concentration and timing on Gram-positive and Gram-negative bacterial classes. The results indicated that among the tested AOFs, Visuprime, Iodim, and Oftasteril were the most rapid and effective for ocular surface disinfection against the tested bacterial species. Conclusions: The obtained results can support the clinician’s choice of the most suitable AOF for the prevention and treatment of ophthalmic infections associated with surgery.

First Report of Fusarium proliferatum Causing Pinellia ternata Corm Rot in Henan, China.

Pinellia ternata (Thunb.) Makino is a perennial herb of the genus Pinellia in the Araceae family, which is a kind of precious Chinese herbal medicine (Bai et al. 2022). In May 2023, rotting was observed on P. ternata corm in the Tanghe planting base (3.33 ha, 32º46´6˝N, 113º 8´47˝E, 138 m), with an incidence of approximately 10%. To identify the causal agent, five rotten corms were randomly sampled from the Tanghe planting base. Symptomatic corm tissues were cut into small pieces (3 mm3) with asymptomatic tissue margins, disinfected in 75% ethanol for 30s and in 2% sodium hypochlorite for 3 min, rinsed three times with sterile distilled water, and then placed on synthetic nutrient-poor agar (SNA) plates incubated at 20℃ with a 12 h light/dark photoperiod. Hyphal-tips from the growing edge of colonies were transferred to fresh SNA to obtain pure cultures. In total, five pure isolates were obtained with similar colony morphology and white (top view) or grayish white (back view) coloration, with villous surfaces and abundant aerial hyphae. On SNA, the isolates mainly produced microconidia. Microconidia were ellipsoidal to cylindrical, thin-walled, smooth, hyaline and measured 5.40 to 11.98 (av. 7.14) µm long and 1.60 to 5.38 (av. 2.54) µm thick (n=50). Based on these morphological characteristics of colony and conidia, the causal agent was preliminarily identified as Fusarium sp. (Crous et al. 2021). To confirm pathogen identity, the total genomic DNA of 2 isolates (BX-2 and BX-3) was extracted using the 2×T5 Direct PCR kit (Beijing Tsingke Biotech Co., Ltd, Beijing, China). The internal transcribed spacer (ITS), RNA polymerase II second largest subunit (rpb2), translation elongation factor 1-alpha (tef1), and β-tubulin (tub2) gene were PCR-amplified using the universal primers ITS1/ITS4, 5f2/7cr, EF-1/EF-2 and BT2F/BT4R (Crous et al. 2021), respectively. Sequences have been deposited in GenBank (PP125560-PP125561 for ITS, PP782474-PP782475 for rpb2, PP782476-PP782477 for tef1, PP171436-PP171437 for tub2). BLAST search showed that all sequences were 99% to 100% homology with the corresponding sequences of Fusarium proliferatum (CBS 182.32). A phylogenetic tree was constructed based on the Maximum-Likelihood method in MEGA-X (Kumar et al. 2018) and revealed that the two isolates were closest to F. proliferatum. The pathogenicity tests were conducted with the two isolates (BX-2 and BX-3) as described by Liu et al. (2022) with slight modifications. For each treatment, five healthy P. ternata corms were inoculated with 1ml of conidial suspension (1×107 conidia/ml). Control plants (n=5) were inoculated with sterilized water. All samples were covered with plastic bags and maintained at a relative humidity greater than 90% in a greenhouse at 20℃ with a 12 h light/dark photoperiod. After 15 days, inoculated corms showed similar symptoms to those on the original diseased plants, while control plants remained symptomless. F. proliferatum was successfully reisolated from the inoculated plants and identified based on its morphological characteristics. Thus, Koch's postulates were fulfilled. The experiment was repeated thrice with similar results. Morphological characteristics, sequence alignment and Koch's postulates test confirmed that F. proliferatum was the cause of P. ternata corm rot disease. To our knowledge, this is the first report of F. proliferatum causing P. ternata corm rot in Henan province, China. Our findings are important for informed surveillance of the disease.

Differentiation complex sputum microbiome in patients suspected TB pulmonary

<b>Purpose: </b>This is the first study to attempt microbiome diversity using metagenomic full-length 16S rRNA from respiratory specimens suspected of chronic pulmonary TB patients.<br /> <b>Materials and methods:</b> A 33 patients with suspected chronic pulmonary TB were included. Sputum specimens were cultured to detect <i>mycobacterium sp.</i> and extracted using QiAmp DNA mini kit modification and 16S rRNA metagenomic sequencing by nanopore grid ion sequencer. Microbiome analysis was performed using Pavian and Krona tools.<br /> <b>Results: </b>9 patients were diagnosed with TB based on GeneXpert MTB/RIF assay, and 3 patients were detected with NTM pulmonary infection.The common genera identified from TB culture positive patients were <i>streptococcus sp.</i>, <i>prevotella sp., </i>and <i>veilonella sp. </i>However, less was detected in two NTM infection patients<i>. </i>Metagenomic analysis revealed community bacteria species, including mycobacterium tuberculosis and NTM species, with the lowest number of unique reads. The abundance of <i>streptococcus sp.</i> were less than 30% in 4 patient with comorbid diabetes mellitus.<br /> <b>Conclusions:</b> Metagenomic targeted 16SrRNA full-length sequencing in the clinical respiratory specimen can provide diagnostic insight beyond standard microbiologic cultures and detailed profiling of microbial communities at the species level.

Canine Brucellosis: A Comprehensive Review

Canine brucellosis, caused by Brucella canis, is a global infectious and zoonotic disease that poses a significant public health risk due to the close interactions between dogs and humans. In dogs, the disease can lead to abortion outbreaks, reproductive failures, lymphadenopathy, and occasionally osteoarticular issues. Asymptomatic infections are also relatively common. In humans, brucellosis typically presents with a febrile illness and non-specific symptoms such as splenomegaly, fatigue, and weakness. The disease is notably problematic in breeding programs due to reproductive failures, including infertility and abortion, leading to substantial economic losses. Transmission occurs through direct contact with infected bodily fluids, particularly during mating, or via contaminated environments. Diagnosing canine brucellosis accurately is critical and can be achieved using various methods, including bacterial cultures, serological tests like the Rose Bengal Plate Test (RBPT) and Enzyme-Linked Immunosorbent Assay (ELISA), and molecular techniques such as polymerase chain reaction (PCR). Treatment is limited, with neutering and antibiotic therapy being the main approaches, though these do not always ensure complete recovery. Preventive measures are essential and include regular screening of breeding dogs, isolation of infected animals, and strict biosecurity protocols. Given its zoonotic potential, it is crucial for veterinary professionals and pet owners to be aware of the disease. Increased awareness and future research on new diagnostic methods and effective vaccines are vital for improving control strategies and reducing the impact of canine brucellosis on both canine and human health.

Production and Antimicrobial Activities of Pyocyanin Extracts from Pseudomonas aeruginosa

Pyocyanin is a blue-green phenazine pigment synthesized by Pseudomonas aeruginosa that has significant biotechnological applications. The present study aims to investigate pyocyanin extracts of multi-drug resistant P. aeruginosa isolates sourced from hospital wastewater and evaluate their antimicrobial activity against a panel of clinically relevant pathogens such as Staphylococcus aureus, Salmonella typhi, Achromobacter xylosoxidans and Candida albicans. Nutrient broth and King’s A broth supplemented with 1% nutrient supplements such as rice water and groundnut cake powder were used as a production medium. Extracted pyocyanin was confirmed by FTIR analysis. The isolates P8 and P9 demonstrated of varying concentrations of pyocyanin in different media. Isolate P8 showed maximum pyocyanin production in King’s A broth compared to nutrient broth with pyocyanin yields 14.34 (µg/mL) and 5.63 (µg/mL), respectively, without the nutrient supplements. Preliminary antimicrobial activity of the pyocyanin extracts exhibited substantial inhibition of tested bacterial culture at a concentration of 25 mg/µl; however, did not show any antifungal activity against tested fungi.

Rapid bacterial identification through volatile organic compound analysis and deep learning.

The increasing antimicrobial resistance caused by the improper use of antibiotics poses a significant challenge to humanity. Rapid and accurate identification of microbial species in clinical settings is crucial for precise medication and reducing the development of antimicrobial resistance. This study aimed to explore a method for automatic identification of bacteria using Volatile Organic Compounds (VOCs) analysis and deep learning algorithms. AlexNet, where augmentation is applied, produces the best results. The average accuracy rate for single bacterial culture classification reached 99.24% using cross-validation, and the accuracy rates for identifying the three bacteria in randomly mixed cultures were SA:98.6%, EC:98.58% and PA:98.99%, respectively. This work provides a new approach to quickly identify bacterial microorganisms. Using this method can automatically identify bacteria in GC-IMS detection results, helping clinical doctors quickly detect bacterial species, accurately prescribe medication, thereby controlling epidemics, and minimizing the negative impact of bacterial resistance on society.

Primary osteosarcoma of the breast during lactation: a case report and literature review

Primary osteosarcoma of the breast (POB) is an aggressive and exceedingly rare tumor, and cases with onset during lactation are extremely rare. The exact mechanism of POB development remains unclear. They may originate from totipotent mesenchymal cells in the breast stroma or may be the result of neoplastic transformation of prior breast lesions. Here, we present the case of a 40-year-old Chinese woman who was found with a palpable tumor measuring approximately 3 cm in diameter in her right breast while breastfeeding 4 months post-partum. The lactating woman was misdiagnosed with lactational mastitis during her initial visit to a community hospital. However, due to negative bacterial cultures and ineffective anti-infective treatment, later on the patient was taken to a more advanced hospital where a tissue biopsy was taken. The superior hospital considered that it might be a malignant tumor and performed local excision of the breast mass, leading to a final pathological diagnosis of POB. This is the first reported case of primary osteosarcoma during breastfeeding. We hope that this case report will improve readers’ understanding of the diagnosis and differential diagnosis of POB, especially for patients with atypical clinical symptoms and imaging findings, which should not be taken lightly.

Optimal incubation duration of liquid cultures for assessing culture negative conversion in patients with Mycobacterium avium complex and Mycobacterium abscessus pulmonary diseases.

Mycobacterial liquid culturing typically requires six weeks or longer, primarily because of the slow growth rate of Mycobacterium tuberculosis. This study aimed to evaluate the potential of shortening the duration of mycobacterial liquid culturing in healthcare settings with high prevalence rates of non-tuberculous mycobacteria. We retrospectively analyzed the relationship between mycobacterial species and time to positive testing of liquid cultures from sputum samples using the Mycobacteria Growth Indicator Tube system over a 3.5-year period beginning in July 2020 at a university hospital in Japan. We analyzed 15,147 sputum culture samples and found a 1.1% positivity rate for Mycobacterium tuberculosis complex, while the rates for Mycobacterium avium complex and Mycobacterium abscessus were 17.6% and 2.1%, respectively. The median time to positivity was 17 days for Mycobacterium tuberculosis complex, 9 days for Mycobacterium avium complex, and 4 days for Mycobacterium abscessus. Comparing a 4-week culture period with an eight-week period, the positivity rates for Mycobacterium avium complex and Mycobacterium abscessus were 97.0% and 99.4%, respectively. In settings with a high incidence of non-tuberculous mycobacteria, the basic liquid culturing period can be safely shortened to 4 weeks without significantly compromising detection sensitivity, except for the samples that are highly suspected to contain tuberculosis, extremely slow-growing mycobacteria, smear-positive, or nucleic acid amplification testing positive.

Application of 3D printing in bacterial detection

Diseases caused by bacterial infections, especially pathogen variations and drug-resistant bacteria, are a serious threat to human health globally. Accurate and rapid identification of bacterial pathogens is essential for early diagnosis of infections and timely treatment of disease. At present, the routine diagnostic methods for identifying bacterial pathogens in clinic are bacterial culture, immunological detection, genetic analysis, etc. These methods occupy foundational position for bacterial detection that favors for clinical diagnosis of pathogen infection in daily life. Notably, 3D printing technology, together with 3D bioprinting technology, provides more options for bacterial detection due to its personalized design and manufacturing. Compared with traditional methods, 3D printing provides a more convenient, rapid and accurate bacterial detection platform, thus meeting the urgent needs of clinical and scientific research. Here, we have summarized the research progress and given a comprehensive review of 3D printing technology in bacterial detection, including bacterial detection sensors, microfluidic chips, bioprinting microarray technology, AI and spectroscopy technology, providing a scientific reference and filling in gaps in 3D printing-assistance bacterial detection. At the same time, technical advantages, challenges and future development trends will also be discussed in related healthcare fields.

Bacterial contamination of autologous blood salvaged during deceased donor liver transplantation: a prospective observational study.

Decompensated cirrhotic patients experience severely increased intestinal permeability and bacterial translocation. Thus, autologous blood salvaged during deceased donor liver transplantation (DDLT) may be contaminated with enteric bacteria. We aimed to evaluate bacterial contamination of autologous blood salvaged during DDLT and its association with post-transplant bacteremia. In 30 patients undergoing DDLT, bacterial culture was performed in salvaged autologous blood samples: one before graft reperfusion (non-leukoreduced) and two after graft reperfusion (non-leukoreduced and leukoreduced). The primary outcome was bacterial contamination of salvaged autologous blood. Seven of 30 patients (23.3%) were positive for bacteria (3 enteric/4 non-enteric) before graft reperfusion while 11 patients (36.7%) were positive (5 enteric/6 non-enteric) after graft reperfusion. Six of 7 patients who were positive for bacteria before graft reperfusion were positive after graft reperfusion with the same bacteria. Only 4 of 11 contaminated blood samples were converted to negative after leukoreduction. Post-transplant bacteremia risk was insignificantly greater in patients who received autologous blood with bacteria than in patients without bacteria (30.0% vs. 5.0%, P = 0.06). We found contamination of salvaged autologous blood with enteric bacteria throughout DDLT and incomplete performance of leukoreduction, indicating high bacterial load. The potential association between contaminated autotransfusion and post-transplant bacteremia warrants further validation in a larger prospective study.Clinical trial notation: This study was registered at the Clinical Research Information Service (CRiS; https://cris.nih.go.kr ; No. KCT0007223; principal registration investigator: Sangbin Han, date of registration: April 25, 2022).

From Killer to Solution: Evaluating Bioremediation Strategies on Microbial Diversity in Crude Oil-Contaminated Soil over Three to Six Months in Port Harcourt, Nigeria

The study aimed to evaluate the efficacy of various bioremediation approaches on microbial diversity in crude oil-contaminated soil over three to six months in Port Harcourt, Nigeria. The objective was to assess the impact of different bioremediation strategies on microbial populations, particularly focusing on hydrocarbon-utilizing bacteria and fungi. Microbial populations were quantified using serial dilution and microbial count techniques. The vapor phase transfer mechanism was employed to estimate hydrocarbon-utilizing bacteria and fungi. Bacterial and fungal colonies were incubated for five days, followed by biochemical tests for isolate identification. Fungal pure cultures were observed under a microscope. The study observed a significant increase in microbial populations in soil free of crude oil pollution when bioremediators such as mushrooms and earthworms were introduced. Mushrooms exhibited a 50% increase in hydrocarbon-utilizing bacteria (HUB), while earthworms showed a 55% increase in HUB over the three to six-month period. The longer lifespan and nutrient absorption capabilities of earthworms facilitated faster growth. Furthermore, significant growth in the microbial population of hydrocarbon-utilizing bacteria and fungi was observed in crude oil-polluted soil after employing bioremediation, with the highest growth observed in soil treated with mushrooms at six months, followed by earthworms at six months. Conversely, the lowest microbial population was recorded in soil polluted with 10% crude oil and remediated with earthworms at three months. The results suggest that mushrooms and earthworms effectively increase microbial populations in crude oil-polluted soil. However, mushrooms demonstrated a higher microbial population increase compared to earthworms, especially in terms of promoting the growth of hydrocarbon-utilizing bacteria (HUB) and hydrocarbon-utilizing fungi (HUF). Based on the findings, it is recommended to prioritize using mushrooms as bioremediation agents in similar environmental restoration efforts due to their superior efficacy in increasing microbial populations, particularly HUB and HUF. This study underscores the potential of mushrooms and earthworms as effective bioremediation agents for restoring microbial diversity in crude oil-contaminated soil, offering insights for sustainable environmental restoration practices in oil-affected regions like Port Harcourt, Nigeria.

Antimicrobial Activity of Diffusible Substances Produced by Lactococcus lactis Against Bacillus cereus in a Non-Contact Co-Culture Model

The symptoms of foodborne illness caused by Bacillus cereus often go unreported, complicating the effectiveness of conventional chemical and physical methods used to inhibit its growth in food production. This challenge, combined with the increasing use of lactic acid bacteria (LAB) in the food industry and consumer preference for minimally processed products, prompted this study. The antibacterial activity of diffusible substances produced by Lactococcus lactis ATCC 11454 against Bacillus cereus NC11143 and Escherichia coli K-12 MG1655 was investigated using a non-contact co-culture model utilising deMann Rogosa and Sharpe broth, with glucose as a carbon source. This study employed plate counting and flow cytometry to assess the impact of these substances on bacterial growth and to analyse their composition and antimicrobial efficacy. The co-culture of Lactococcus lactis ATCC 11454 resulted in the production of a stable antimicrobial peptide, which was heat resistant and acid tolerant. Purification was achieved via ammonium sulphate precipitation and preparative HPLC, yielding a peptide with a molecular mass of 3.3 kDa, with daughter ion fractions similar to nisin A. Antimicrobial activity studies demonstrated that the diffusible substances effectively inhibited B. cereus growth over a period of eight days and exhibited bactericidal activity, killing 99% of the B. cereus cells. Additionally, these substances also inhibited Escherichia coli K-12 MG1655 grown under similar conditions. Comparative analysis revealed that in the co-culture assay, L. lactis produced a 50% higher yield of the antimicrobial peptides compared to pure cultures. Similarly, the specific growth rate of L. lactis was four times higher. With respect to protein purification and concentration, ammonium sulphate precipitation coupled with solid phase extraction was most effective in the purification and concentration of the diffusible substances. The findings provide a basis for utilising bacteriocin-producing strains as a preservation method, offering an alternative to traditional chemical and physical control approaches especially for the food industry.

Molecular Diagnostics for Group A Streptococcal Pharyngitis: Clinical and Economic Benefits in the Belgian Healthcare Context

(1) Background: Group A Streptococcal (GAS) pharyngitis is common, resulting in numerous ambulatory visits. Accurate diagnosis is challenging. This study evaluated the clinical utility, cost, and performance of a nucleic acid amplification test (NAAT) for GAS detection, comparing it to a rapid antigen detection test (RADT) and throat culture. Additionally, we assessed the diagnostic stewardship related to these testing methods to ensure appropriate antibiotic use in clinical practice. Methods: Between November 2022 and February 2023, 82 throat swabs were analyzed, with McIsaac clinical scores calculated for each. The Abbott ID NOW STREP A 2 NAAT and Sekisui Diagnostics’ OSOM® STREP A RADT were performed, followed by bacterial culture. Diagnostic performance was compared using culture as the gold standard. Results: Of the 82 samples, 28 (34.14%) tested positive for pathogenic germs, primarily Streptococcus pyogenes (92.85%). RADTs showed a sensitivity of 80.76% and a specificity of 100%, while NAATs demonstrated a sensitivity of 100% and specificity of 96.42%. Cost analysis indicated the need for reimbursement adjustments to optimize NAAT’s economic benefits. Clinical data indicated that symptoms alone were insufficient for reliable diagnosis. Conclusions: This study confirmed the superior sensitivity of Abbott’s Strep A2 NAAT over RADT. Given the Belgian guidelines against routine antibiotic treatment for pharyngitis and considering local treatment recommendations and cost, implementing NAAT for GAS detection in Belgian laboratories is less beneficial. However, the role of NAAT in supporting antimicrobial stewardship by ensuring appropriate antibiotic use remains significant.

Microbiological surveillance result of endoscopes after INTERCEPT Foam Spray: a quasi-experimental pilot study in Singapore.

This study aimed to assess the impact of INTERCEPT Foam Spray (IFS) application on delayed endoscope reprocessing through microbiological surveillance culture (MSC). A quasi-experimental, matched-comparison pilot study was conducted using gastrointestinal endoscopy. IFS was applied to the endoscopes after precleaning and before reprocessing the next day. An equal number of endoscopes, matched by endoscope type, were subjected to routine reprocessing. The MSC were subjected to high-level disinfection to detect any contamination. Data were analyzed using the chi-square test or Fisher exact test (categorical data) and Student t-test (continuous data). In total, 150 MSCs were collected from 42 endoscopes. Positive MSCs were observed in 4.0% (4/75) of the sprayed group and 1.3% (1/75) of the control group (95% confidence interval [CI], 30.34-0.31; p>0.05), all of which were contributed by colonoscopes. Colonoscope were more prone to positive MSC (mean difference in percentage, p<0.05). Mean spraying hours were not associated with detected growth (11.7% vs. 13.6; 95% CI, 1.43 to -5.27; p>0.05), with environmental and skin flora being the primary contaminants. IFS may be applied when delayed endoscope processing is necessary, but with caution when applied to colonoscopes. However, further research is warranted to verify the result.

Comparison of Four Different DNA Isolation Methods from MGIT Culture for Long-Read Whole Genome Sequencing of Mycobacterium tuberculosis

Background: Tuberculosis (TB) remains a global health challenge, particularly due to drug resistance and limitations in rapid diagnosis. Next-generation sequencing (NGS), especially long-read whole genome sequencing (WGS), shows promise for rapidly detecting TB and drug resistance, but it requires high-quality DNA, which is difficult to extract from Mycobacterium tuberculosis due to its complex cell wall. Objectives: This study evaluated four DNA isolation methods for extracting pure DNA from M. tuberculosis, aiming to standardize protocols for long-read WGS. Methods: Mycobacterium tuberculosis H37RV colonies were grown in BACTEC MGIT liquid medium. Two pellets were prepared as the initial material for the DNA extraction protocol: Pellets from 1 mL McFarland 2 suspensions and all growing colonies from two MGIT liquid cultures. Four DNA extraction methods were used: The cetyltrimethylammonium bromide (CTAB) method, GeneJET Genomic DNA Purification Kit, Quick-DNA Fecal/Soil Microbe Kit, and Genematrix Tissue/Bacterial DNA Purification Kit, with some modifications. DNA quality was assessed based on concentration, purity, and integrity. Results: Among the tested methods, the Quick-DNA Fecal/Soil Kit yielded approximately 85 ng/mL of DNA and a purity of 1.9 at 260/280 nm from the colonial pellet of two MGIT tubes. However, lower intact DNA [DNA integrity number (DIN) ~ 6.8] was obtained with this kit. The CTAB method provided the highest intact DNA (DIN ~ 9.5), although the purity of the DNA was not sufficient. Conclusions: Based on three repetitions of McF-2 and colonial pellet extractions, the Quick-DNA Fecal/Soil Kit yielded the highest DNA quantity and purity but showed lower integrity compared to other methods, indicating the need for adjustments. A pellet from two MGIT cultures (~ 100 µL) is suitable for long-read WGS with this kit. However, a larger sample size is required to generalize these findings. For effective long-read sequencing of M. tuberculosis, DNA extraction protocols must be optimized to balance yield, fragment size, and purity for accurate sequencing and drug resistance analysis.

The clinical management and efficacy of metagenomic next-generation sequencing in patients with pyogenic spinal infection: a single-center cohort study

ObjectiveThis study aims to evaluate the clinical management and effectiveness of metagenomic next-generation sequencing (mNGS) in patients with pyogenic spinal infections.MethodsWe conducted a retrospective review of 17 patients diagnosed with pyogenic spinal infections and treated at our institution between October 2022 and February 2024. The cohort included 8 males and 9 females, with a mean age of 63.59 ± 10.18 years (range: 41–71 years). The infections comprised 9 epidural abscesses, 6 intervertebral space infections, and 2 deep abscesses. All patients underwent open surgical procedures and mNGS-based bacterial identification using intraoperative pus or tissue specimens, in addition to conventional blood bacterial cultures. Clinical outcomes were assessed using CRP, PCT, WBC inflammatory markers, and VAS scores postoperatively.ResultsAll 17 patients with pyogenic spinal infections underwent open surgery and mNGS bacterial detection at our institution. Among the 17 patients, mNGS yielded positive results in 14 cases (82.4%), significantly higher than the 5.9% positivity rate of conventional bacterial cultures (p < 0.001). The mNGS test time was notably shorter than conventional cultures (1.0 vs. 5.88 days, p < 0.001). Postoperative antibiotic therapy was adjusted based on mNGS findings. There were significant reductions in postoperative VAS, WBC, PCT, and CRP values compared to preoperative levels (p < 0.01).ConclusionMetagenomic next-generation sequencing is effective in managing pyogenic spinal infections by facilitating rapid and sensitive detection of pathogens. This technique improves the timeliness and accuracy of diagnosis, highlighting its potential for broader clinical use.

Lumbosacral rotation flap: a simple method for covering sacral pressure injuries.

Pressure injuries (PIs) are among the most common skin and soft tissue wounds occurring in patients who are bedbound and/or immobile. PI management hinges on their prevention; however, reoccurrence poses a challenge to their management and requires a multidisciplinary approach. Here, the authors describe a lumbosacral rotation flap (LSRF) for the coverage of sacral PIs. A single-centre, retrospective analysis of prospectively collected data was carried out. All patients undergoing LSRF for sacral PIs were included. Patients with active systemic sepsis, immune compromise, hepatic or renal dysfunction were excluded. All patients underwent preoperative optimisation and wound cultures to direct antibiotic therapy after surgery. A total of nine patients underwent the procedure (seven male and two female). Mean age was 47.6 years with a mean ulcer size of 92.9 cm2. Bone biopsy indicated the presence of osteomyelitis in three patients. Of the LSRFs, two flaps showed minimal local complications in the form of marginal flap necrosis which was managed conservatively. All flaps healed well with no cases of flap loss or the need for secondary procedures. The results of this analysis showed that LSRF can be considered a first line of treatment of sacral PIs. They can be used to cover large defects. Due to their large base and flap size, readvancement in cases of recurrence is also possible.