Bronchoalveolar Lavage Fluid Culture Research Articles

Case report: Rare pulmonary fungal infection caused by Penicillium digitatum: the first clinical report in China

Penicillium digitatum is a common plant pathogen that causes citrus rot, which is extremely rare in humans. We report a case of a 66-year-old man with a history of consuming large amounts of citrus fruits, smoking for 30 years, and a history of emphysema. He had experienced intermittent coughing with sputum for more than 10 years and was admitted to the hospital due to worsening of symptoms over the past month. Despite antibiotic treatment, his condition did not improve. Subsequently, bronchoalveolar lavage fluid (BALF) was detected by metagenomic next-generation sequencing (mNGS), which showed the presence of P. digitatum. The fungal culture of BALF also indicated the presence of the Penicillium genus. The diagnosis was lung infection caused by P. digitatum, and the patient was treated with itraconazole. The lung infection was controlled. This is the third reported case of invasive pulmonary fungal infection caused by P. digitatum worldwide at the genus level, and the first reported case in China. Although human infections caused by P. digitatum are rare, as an emerging opportunistic pathogen, the detection of this fungus in immunocompromised patients should still be clinically important.

A rare family outbreak of Mycobacterium abscessus infection in immunocompetent fraternal triplets

BackgroundMycobacterium abscessus (M. abscessus) infection is rare in children who were previously healthy, particularly in infants. We present the first report of a family outbreak of M. abscessus infection among immunocompetent infant triplets. MethodsWe reviewed triplets’ demographic data, laboratory tests and imaging examinations to describe their clinical features. We performed whole-exome sequencing to rule out primary immunodeficiency disorders. We used DNA sequencing for M. abscessus subspecies identification. ResultsThe fraternal triplets (triples A, B and C) presented with a 10-day history of cough. Triple A also experienced a brief episode of fever, and triple B had tachypnea. Chest CT scans showed pulmonary masses and nodules in triples A and C, and cavities in triple B. Cultures of sputum and bronchoalveolar lavage fluid from all triplets yielded M. abscessus. Further subspecies identification showed that isolates from triples A and C were M. abscessus subsp. massiliense, and isolates from triple B were M. abscessus subsp. abscessus (MAA). After eight months of combination therapy, the pulmonary lesions of the triplets improved significantly. ConclusionOur study confirms that M. abscessus pulmonary disease can occur in immunocompetent infants. We hypothesize that the simultaneous infection of the triplets may be associated with their prematurity and extensive environmental exposure. This study highlights the importance to include M. abscessus infection in the differential diagnosis of pulmonary masses and/or cavities, regardless of the age of onset or the presence of underlying pathology or susceptible genes.

Performance of rapid on-site evaluation of touch imprints of bronchoscopic biopsies or lung tissue biopsies for the diagnosis of invasive pulmonary filamentous fungi infections in non-neutropenic patients.

The diagnosis of invasive pulmonary fungal disease depends on histopathology and mycological culture; there are few studies on touch imprints of bronchoscopic biopsies or lung tissue biopsies for the diagnosis of pulmonary filamentous fungi infections. The purpose of the present study was to explore the detection accuracy of rapid on-site evaluation of touch imprints of bronchoscopic biopsies or lung tissue biopsies for the filamentous fungi, and it aims to provide a basis for initiating antifungal therapy before obtaining microbiological evidence. We retrospectively analyzed the diagnosis and treatment of 44 non-neutropenic patients with invasive pulmonary filamentous fungi confirmed by glactomannan assay, histopathology, and culture from February 2017 to December 2023. The diagnostic positive rate and sensitivity of rapid on-site evaluation for these filamentous fungi identification, including diagnostic turnaround time, were calculated. Compared with the final diagnosis, the sensitivity of rapid on-site evaluation was 81.8%, and the sensitivity of histopathology, culture of bronchoalveolar lavage fluid, and glactomannan assay of bronchoalveolar lavage fluid was 86.4%, 52.3%, and 68.2%, respectively. The average turnaround time of detecting filamentous fungi by rapid on-site evaluation was 0.17 ± 0.03 hours, which was significantly faster than histopathology, glactomannan assay, and mycological culture. A total of 29 (76.3%) patients received earlier antifungal therapy based on ROSE diagnosis and demonstrated clinical improvement. Rapid on-site evaluation showed good sensitivity and accuracy that can be comparable to histopathology in identification of pulmonary filamentous fungi. Importantly, it contributed to the triage of biopsies for further microbial culture or molecular detection based on the preliminary diagnosis, and the decision on early antifungal therapy before microbiological evidence is available.

A complex case study: coexistence of multi-drug-resistant pulmonary tuberculosis, HBV-related liver failure, and disseminated cryptococcal infection in an AIDS patient

BackgroundHepatitis B virus (HBV) infection can cause liver failure, while individuals with Acquired Immunodeficiency Virus Disease (AIDS) are highly susceptible to various opportunistic infections, which can occur concurrently. The treatment process is further complicated by the potential occurrence of immune reconstitution inflammatory syndrome (IRIS), which presents significant challenges and contributes to elevated mortality rates.Case presentationThe 50-year-old male with a history of chronic hepatitis B and untreated human immunodeficiency virus (HIV) infection presented to the hospital with a mild cough and expectoration, revealing multi-drug resistant pulmonary tuberculosis (MDR-PTB), which was confirmed by XpertMTB/RIF PCR testing and tuberculosis culture of bronchoalveolar lavage fluid (BALF). The patient was treated with a regimen consisting of linezolid, moxifloxacin, cycloserine, pyrazinamide, and ethambutol for tuberculosis, as well as a combination of bictegravir/tenofovir alafenamide/emtricitabine (BIC/TAF/FTC) for HBV and HIV viral suppression. After three months of treatment, the patient discontinued all medications, leading to hepatitis B virus reactivation and subsequent liver failure. During the subsequent treatment for AIDS, HBV, and drug-resistant tuberculosis, the patient developed disseminated cryptococcal disease. The patient’s condition worsened during treatment with liposomal amphotericin B and fluconazole, which was ultimately attributed to IRIS. Fortunately, the patient achieved successful recovery after appropriate management.ConclusionEnhancing medical compliance is crucial for AIDS patients, particularly those co-infected with HBV, to prevent HBV reactivation and subsequent liver failure. Furthermore, conducting a comprehensive assessment of potential infections in patients before resuming antiviral therapy is essential to prevent the occurrence of IRIS. Early intervention plays a pivotal role in improving survival rates.

Microbiological Rapid On-Site Evaluation for Pulmonary Infectious Diseases.

The prompt initiation of empirical anti-infective therapy is crucial in patients presenting with unexplained pulmonary infection. Although imaging acquisition is relatively straightforward in clinical practice, its lack of specificity often necessitates additional time-consuming tests such as sputum culture, bronchoalveolar-lavage fluid culture, or genetic sequencing to identify the underlying etiology of the disease accurately. Moreover, the limited efficacy of empirical anti-infective treatment may contribute to antibiotic misuse. Recent advancements in interpreting microbial background on rapid on-site evaluation (ROSE) slides have enabled clinicians to promptly obtain samples through bronchoscopy (e.g., alveolar lavage, mucosal brushing, tissue clamp), facilitating bedside staining and interpretation that provides essential microbial background information. Consequently, this establishes a foundation for developing targeted anti-infection treatment and individualized drug therapy plans. With a better understanding of which pathogens are causing infections in real-time, physicians can avoid unnecessary broad-spectrum antibiotics contributing to antibiotic resistance. Establishing a rapid and standardized M-ROSE workflow within respiratory medicine departments or intensive care units will greatly assist physicians in formulating accurate treatment strategies for patients, which holds significant clinical implications.

Association between Clinical Characteristics and Microbiota in Bronchiectasis Patients Based on Metagenomic Next-Generation Sequencing Technology.

This study aimed to investigate the disparities between metagenomic next-generation sequencing (mNGS) and conventional culture results in patients with bronchiectasis. Additionally, we sought to investigate the correlation between the clinical characteristics of patients and their microbiome profiles. The overarching goal was to enhance the effective management and treatment of bronchiectasis patients, providing a theoretical foundation for healthcare professionals. A retrospective survey was conducted on 67 bronchiectasis patients admitted to The First Hospital of Jiaxing from October 2019 to March 2023. Clinical baseline information, inflammatory indicators, and pathogen detection reports, including mNGS, conventional blood culture, bronchoalveolar lavage fluid (BALF) culture, and sputum culture results, were collected. By comparing the results of mNGS and conventional culture, the differences in pathogen detection rate and pathogen types were explored, and the diagnostic performance of mNGS compared to conventional culture was evaluated. Based on the various pathogens detected by mNGS, the association between clinical characteristics of bronchiectasis patients and mNGS microbiota results was analyzed. The number and types of pathogens detected by mNGS were significantly larger than those detected by conventional culture. The diagnostic efficacy of mNGS was significantly superior to conventional culture for all types of pathogens, particularly in viral detection (p < 0.01). Regarding pathogen detection rate, the bacteria with the highest detection rate were Pseudomonas aeruginosa (17/58) and Haemophilus influenzae (11/58); the fungus with the highest detection rate was Aspergillus fumigatus (10/21), and the virus with the highest detection rate was human herpes virus 4 (4/11). Differences were observed between the positive and negative groups for P. aeruginosa in terms of common scoring systems for bronchiectasis and whether the main symptom of bronchiectasis manifested as thick sputum (p < 0.05). Significant distinctions were also noted between the positive and negative groups for A. fumigatus regarding Reiff score, neutrophil percentage, bronchiectasis etiology, and alterations in treatment plans following mNGS results reporting (p < 0.05). Notably, 70% of patients with positive A. fumigatus infection opted to change their treatment plans. The correlation study between clinical characteristics of bronchiectasis patients and mNGS microbiological results revealed that bacteria, such as P. aeruginosa, and fungi, such as A. fumigatus, were associated with specific clinical features of patients. This underscored the significance of mNGS in guiding personalized treatment approaches. mNGS could identify multiple pathogens in different types of bronchiectasis samples and was a rapid and effective diagnostic tool for pathogen identification. Its use was recommended for diagnosing the causes of infections in bronchiectasis patients.

Clinical application of bronchoalveolar lavage fluid metagenomics next-generation sequencing in cancer patients with severe pneumonia

ObjectiveMetagenomic next-generation sequencing (mNGS), as an emerging technique for pathogen detection, has been widely used in clinic. However, reports on the application of mNGS in cancer patients with severe pneumonia remain limited. This study aims to evaluate the diagnostic performance of bronchoalveolar lavage fluid (BALF) mNGS in cancer patients complicated with severe pneumonia.MethodsA total of 62 cancer patients with severe pneumonia simultaneously received culture and mNGS of BALF were enrolled in this study. We systematically analyzed the diagnostic significance of BALF mNGS. Subsequently, optimization of anti-infective therapy based on the distribution of pathogens obtained from BALF mNGS was also assessed.ResultsFor bacteria and fungi, the positive detection rate of mNGS was significantly higher than culture method (91.94% versus 51.61%, P < 0.001), especially for poly-microbial infections (70.97% versus 12.90%, P < 0.001). Compared with the culture method, mNGS exhibited a diagnostic sensitivity of 100% and a specificity of 16.67%, with the positive predictive value (PPV) and negative predictive value (NPV) being 56.14% and 100%, respectively. The agreement rate between these two methods was 59.68%, whereas kappa consensus analysis indicated a poor concordance (kappa = 0.171). After receipt of BALF mNGS results, anti-infective treatment strategies in 39 out of 62 cases (62.90%) were optimized. Moreover, anti-tumor therapy was a high-risk factor for mixed infections (87.18% versus 65.22%, P = 0.04).ConclusionsThe present study showed that cancer patients with severe pneumonia, especially those received anti-tumor therapy, were more likely to have poly-microbial infections. BALF mNGS can provide a rapid and comprehensive pathogen distribution of pulmonary infection, making it a promising technique in clinical practice, especially for optimizing therapeutic strategies for cancer patients.

Metagenomics for the microbiological diagnosis of hospital-acquired pneumonia and ventilator-associated pneumonia (HAP/VAP) in intensive care unit (ICU): a proof-of-concept study

BackgroundHospital-acquired and ventilator-associated-pneumonia (HAP/VAP) are one of the most prevalent health-care associated infections in the intensive care unit (ICU). Culture-independent methods were therefore developed to provide faster route to diagnosis and treatment. Among these, metagenomic next-generation sequencing (mNGS) has shown considerable promise.MethodsThis proof-of-concept study describes the technical feasibility and evaluates the clinical validity of the mNGS for the detection and characterization of the etiologic agents causing hospital-acquired and ventilator-associated pneumonia. We performed a prospective study of all patients with HAP/VAP hospitalized in our intensive care unit for whom a bronchoalveolar lavage (BAL) was performed between July 2017 and November 2018. We compared BAL fluid culture and mNGS results of these patients.ResultsA total of 32 BAL fluids were fully analyzed. Of these, 22 (69%) were positive by culture and all pathogens identified were also reported by mNGS. Among the culture-positive BAL samples, additional bacterial species were revealed by mNGS for 12 patients, raising the issue of their pathogenic role (colonization versus coinfection). Among BALF with culture-negative test, 5 were positive in mNGS test.ConclusionsThis study revealed concordant results for pneumonia panel pathogens between mNGS and culture-positive tests and identified additional pathogens potentially implicated in pneumonia without etiologic diagnosis by culture. mNGS has emerged as a promising methodology for infectious disease diagnoses to support conventional methods. Prospective studies with real-time mNGS are warranted to examine the impact on antimicrobial decision-making and clinical outcome.

The Prognosis of Patients Tested Positive for Stenotrophomonas maltophilia from Different Sources.

The purpose of this study was to analyze the prognosis of patients tested positive for Stenotrophomonas maltophilia (SMA) from different sources. A retrospective study was conducted among 651 patients tested positive for SMA from January 2020 to October 2022 in the First Affiliated Hospital of Anhui Medical University. The patients were divided into seven groups by the source of SMA. Univariate and multivariate analyses were used to identify risk factors for mortality in patients tested positive for SMA from different sources. A total of 651 SMA isolates were collected from various sources, including sputum (348 isolates, 53%), bronchoalveolar lavage fluid (52, 8%), abdominal drainage fluid (76, 12%), wound secretion (66, 10%), blood (62, 10%), urine (41, 6%) and cerebrospinal fluid (6, 1%). Compared with other groups, the mortality of the patients in the bronchoalveolar lavage fluid culture group, blood culture group, and abdominal drainage fluid culture group was higher, at 40.38%, 32.26%, and 26.32%, respectively. Multivariate analysis showed that continuous renal replacement therapy was an independent risk factor for mortality in patients with SMA bloodstream infection (P=0.020, OR=6.86), and effective antimicrobial therapy after being positive for S. maltophilia isolates (P=0.002, OR=0.10) was negatively correlated with the death of patients with SMA bloodstream infection. Age ≥65 years (P= 0.043, OR=4.96), kidney disease (P=0.045, OR=4.62) and antifungal agent exposure (P=0.036, OR=5.13) were independent risk factors for mortality in patients with SMA intra-abdominal infection. Antifungal agent exposure (P=0.024, OR=0.51) and glycopeptide exposure (P=0.045, OR=0.53) were independent risk factors for mortality in patients in the sputum culture group. SMA has a high rate of antimicrobial resistance and can cause multisite infection. Pulmonary infections, bloodstream infections and abdominal infections caused by SMA have high mortality, and timely standardized treatment can reduce mortality.

Clinical features and pathogen distributions of microbiological-based protracted bacterial bronchitis in children of different ages in Northeast China.

Protracted bacterial bronchitis (PBB) is often diagnosed clinically according to chronic wet cough, which can be resolved by appropriate antibiotics. Though rarely performed in PBB diagnosis, bacterial cultures by sputum or bronchoalveolar lavage (BAL) fluid can provide etiological features, which may be different in western countries and different areas of China. This study aimed to investigate the clinical and etiological features and outcomes in children of different ages with PBB in northeast China. We retrospectively analyzed children diagnosed with PBB by positive BAL fluid or sputum bacterial cultures between 2017 and 2021. Children were divided into three age groups: <1 year (infants), 1-5 years (younger children), and ≥6 years (older children). Clinical characteristics, chest radiographic findings, bronchoscopy findings, microbiological findings, treatment strategies, and outcomes were reviewed and compared among the age groups. Factors associated with remission during follow-up were examined using logistic regression. A total of 45 children with PBB were included, consisting of 24 (53.3%) infants. The infants were often boys and had a shorter cough duration, a lower proportion of expectoration, a greater proportion of wheezing, and less bronchial wall thickening on high-resolution computed tomography compared to older children (P < 0.05). No significant differences were found among the age groups regarding macroscopic findings, except for a higher proportion of tracheobronchial malacia in infants than in older children (P = 0.013). The most commonly cultured bacteria were Haemophilus influenzae (42.2%), followed by Streptococcus pneumoniae (22.2%) and Klebsiella pneumoniae (20.0%). Compared to older children, infants had a higher remission (P = 0.009) and relatively lower relapse rates (P = 0.059). Short duration of cough (OR = 0.58, 95% CI: 0.34-0.99, P = 0.046) and absence of recurrent cephalosporins before diagnosis (OR = 0.05, 95% CI: 0.00-0.73, P = 0.028) were associated with remission. Infants are more prone to PBB, with increased wheezing. Gram-negative bacilli infections are common in infants in northeast China. Older children with PBB should be carefully assessed, treated and followed up, particularly those with long duration of cough and poor response to antibiotic treatments.

Advancing Microbe Detection for Lower Respiratory Tract Infection Diagnosis and Management with Metagenomic Next-Generation Sequencing.

Due to limitations of traditional microbiological methods and the presence of the oropharyngeal normal flora, there are still many pathogens that cause lower respiratory tract infections (LRTIs) cannot be detected. Metagenomic next-generation sequencing (mNGS) has the potential capacity to solve this problem. This retrospective study successively reviewed 77 patients with LRTI and 29 patients without LRTI admitted to Tianjin Medical University General Hospital, China from August 2020 to June 2021. Pathogens in bronchoalveolar lavage fluid (BALF) specimens were detected adopting mNGS and traditional microbiological assays. The diagnostic performance of pathogens was compared between mNGS and BALF culture. The value of mNGS for aetiological and clinical impact investigation in LRTI was also evaluated. Among 77 patients with LRTI, 22.1%, 40.3%, and 65.0% of cases were detected as definite or probable pathogens by culture, all conventional microbiological tests, and mNGS, respectively. Using the final diagnosis as a gold standard, mNGS exhibited a sensitivity of 76.6% (95% confidence interval [CI], 65.6-85.5%), which was considerably superior to that of BALF culture (76.6% vs 18.2%; P < 0.01); specificity of 79.3% (95% CI, 60.3-92.0%), which was similar (79.3% vs 89.7%; P = 0.38); positive-predictive value of 90.8% (95% CI, 81.0-96.5%), and negative-predictive value of 56.1% (95% CI, 39.7-71.5%). According to our data, mNGS identified potential microorganisms in 66.7% (42/63) of culture-negative samples. Among 59 patients with pathogens identified by mNGS, conventional microbiological methods confirmed pathogenic infections in less than half (28/59) cases. Within the 77 patients, 34 (44.2%) patients received pathogen-directed therapy, 7 (9.1%) patients underwent antibiotic adjustment, and 3 (3.9%) patients stopped using antibiotics due to mNGS results. mNGS exhibits high accuracy in diagnosing LRTI, and combine with traditional microbiological tests, causative pathogens can be detected in approximately 70.0% of cases, thus yields a positive effect on antibiotic application.

Comparison of Third-Generation Sequencing Technology and Traditional Microbiological Detection in Pathogen Diagnosis of Lower Respiratory Tract Infection.

It is common to obtain a low detection rate and unsatisfactory detection results in complex infection or rare pathogen detection. This retrospective study aimed to illustrate the application value and prospect of the third-generation sequencing technology in lower respiratory tract infection disease. This study recruited 70 patients with lower respiratory tract infection (LRTI). Pathogen detection of bronchoalveolar lavage fluid (BALF) from all patients was performed using nanopore metagenomic sequencing technology and traditional culture. BALF culture combined with quantitiative PCR (qPCR) was used as a reference standard to analyze the sensitivity and specificity of nanopore sequencing technology. The current study also collected the examination results of enrolled samples using technical methods sputum culture, tuberculosis DNA (TB-DNA), and Xpert MTB/RIF and analyzed the detection efficiency of nanopore sequencing for Mycobacterium tuberculosis. The positive rates of pathogens in 70 BALF samples detected by conventional culture and nanopore sequencing were 25.71% and 84.29%, respectively. Among the 59 positive BALF cases using nanopore sequencing, a total of 31 pathogens were identified, of which the proportions of bacteria, fungi, viruses, and other pathogens were 50%, 17%, 32%, and 1%, respectively. Using the results combined with culture and qPCR detection methods as the standard, the pathogen detection of BALF using nanopore sequencing had a sensitivity of 70% and a specificity of 91.7%. Additionally, the positive rate of the detection of M. tuberculosis using nanopore sequencing was 33.3% (6/18). The clinical medication plans of 74.3% (52/70) of the patients were referred to the nanopore sequencing results, of which 31 cases changed their treatment strategy, 21 supported the previous treatment plans, and 90% (47/52) of the patients finally had clinical improvement. BALF detection using nanopore sequencing technology improves the process of detecting pathogens in patients with LRTI, especially for M. tuberculosis, fungi, and viruses, by reducing the report time from three days to six hours. The clinical application prospect of nanopore sequencing technology is promising in the pathogen diagnosis of LRTI.

Respiratory Diseases and Their Diagnosis in Dogs: A Review

Respiratory affections are common in dogs. Very young and older dogs are at greater risk as compared to the adult ones. Constant exposure of the respiratory tract to the infectious agents, reaching the upper and lower respiratory tract aerogenously or hematogenously, makes the respiratory system vulnerable to injury. Clinical signs mainly seen in upper respiratory tract diseases involves nasal discharge (serous, mucopurulent or hemorrhagic), sneezing, facial deformity, stertor, lethargy, inappetence and rarely, central nervous system signs whereas additional signs present in lower tract diseases range from dyspnoea, costal or abdominal respiration, cough, nasal discharge and congestion, edema, consolidation of lungs and weight loss. Advanced tests involved in diagnosis of respiratory problems in veterinary practicpe are rhinoscopy, bronchoscopy, transthoracic fine needle aspiration cytology (FNAC) transtracheal wash and bronchoalveolar lavage fluid cytology and culture. This review discusses the causes of upper and lower respiratory tract disorders of canines and various diagnostic tests involved in their diagnosis.

270. Secondary bacterial infection in hospitalized COVID-19 patients. Experience in Colombia, South America

Abstract Background Secondary infections are common among severe COVID-19 patients, increasing complications and mortality risk. These infections are not well characterized in Latin America and the Caribbean. Methods A cross-sectional observational study of adult patients with COVID-19 admitted to the hospital Fundación Valle del Lili in Cali-Colombia from March 2020 to March 2021. Demographic data, clinical characteristics, laboratory parameters, and clinical outcomes were collected. We describe secondary infection, antibiotic therapy, and antibiotic resistance profiles. Secondary infection was defined if the diagnosis occurred ≥48 hours after hospital admission for COVID-19. Results A total of 2138 patients with COVID-19 were analyzed; 350 (16.3%) presented secondary infection. 60% were male; the median age was 65 years [IQR: 55-72]. Glucocorticoid therapy was indicated in 335 patients (96.3%). 281 received high doses and 54 low doses. Bacterial infections were the most common, affecting 81.3 % of patients, followed by fungal (14.4%) and viral (4.3%) infections. Most bacterial isolates were orotracheal secretion, blood, urine, and bronchoalveolar lavage fluid culture. The three most frequently identified bacteria were Klebsiella pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa. Most of the initial isolates were not antibiotic-resistant (75-89.7%). Empiric antibiotic therapy was indicated in 346 patients (98.9%), 268 received carbapenems (76.6%), 267 Vancomycin (76.3%), and 233 cefepime (66.6%). Of the 350 patients, 327 (93.4%) required management in the intensive care unit, and overall mortality was 35.4% (124/350). Conclusion Our results showed a lower frequency of secondary infection than previous reports; However, a high frequency of broad-spectrum antibiotics usage was found despite a high prevalence of non-resistant bacteria. Further studies are needed to establish the best approach for antibiotics therapy. Disclosures All Authors: No reported disclosures.

Concordance between Microorganisms Isolated from Subglottic Secretions and Bronchoalveolar Lavage Fluid in Patients with Ventilator Associated Pneumonia in Intensive Care Unit

Introduction: Nosocomial pneumonia is a common complication in critically ill patients. Mechanical ventilation has been consistently identified as the greatest risk factor for the development of nosocomial pneumonia. Aims: The purpose of the current study is to find the concordance between microorganisms cultured from subglottic secretions and bronchoalveolar lavage fluid (BALF) and to assess the effectiveness of subglottic suctioning for prevention of ventilator-associated pneumonia (VAP). Patients and Methods: This was a prospective, interventional, cohort study. Patients between 15 and 85 years who developed VAP after receiving mechanical ventilation were enrolled in the study. They were randomly divided into two groups: Group No Subglottic Secretion Drainage (NSSD) and Group Continuous Subglottic Secretion Drainage (CSSD). Results: The mean age of the patients was 56 ± 13 years. Fifty-eight percent were males and 42% were females. The incidence of VAP among NSSD group was 41.7% and in CSSD group was 25%. The coefficient of correlation was calculated for the microorganisms cultured in subglottic and BALF using Pearson's correlation. The correlation coefficient was 0.78 for the entire population (P = 0.01). In the NSSD group, it was 0.82 (P = 0.01), and in the CSSD group, it was 0.76 (P = 0.01). The mean Intensive Care Unit (ICU) stay is maximum in NSSD group (12.9 ± 1.84 days) compared to CSSD group (9.2 ± 1.17 days). Conclusion: The concordance of organism grown in subglottic secretions and BALF culture is higher in NSSD group as compared to CSSD group. CSSD is an effective means of reducing the number of ventilator days and ICU stay.

Disseminated nontuberculous mycobacterial infection with cryptic immunodeficiency mimicking malignancy: a case report

BackgroundNontuberculous mycobacteria (NTM) usually invades vulnerable hosts. Disseminated NTM (dNTM) infection can affect nearly all organs and be easily misdiagnosed as metastatic carcinoma or other systemic diseases, especially in seemingly immunocompetent hosts. Identification of underlying immunodeficiency is critical for the diagnosis and treatment of dNTM. Adult-onset immunodeficiency (AOID) with anti-IFN-γ autoantibodies has recently been recognized as a crucial but frequently neglected risk factor for dNTM infection. Frequent relapses of infection are common in AOID patients despite appropriate anti-infective treatment and B-cell-depleting therapy has shown some promising results. Herein, we report a case of dNTM infection mimicking malignancy in an AOID patient who was successfully treated with rituximab.Case presentationA middle-aged male presented with fever, productive cough, multifocal skin abscesses and multiple osteolytic lesions with pathological fractures. Chest CT revealed consolidation of the lingula while bronchoscopy showed a mass completely blocking the airway opening of the inferior lingual segment. Metagenomic next-generation sequencing and mycobacterial culture of skin pus and bronchoalveolar lavage fluid reported Mycobacterium Colombiense, confirming the diagnosis of dNTM infection. However, anti-NTM antibiotics alone failed to prevent disease relapse and progression. Further evaluation indicated undetectable serum IFN-γ concentration and high-titer autoantibodies against IFN-γ, suggesting that AOID was the underlying reason for dNTM. Rituximab was added to treatment and successfully controlled the infection without relapse at one-year follow-up.ConclusionWe reported a rare case of disseminated Mycobacterium Colombiense infection manifested with pulmonary mass, pathological fracture and dermapostasis in a host with AOID. Our case demonstrated that AOID should be screened when patients get the episode of disseminated NTM infection particularly when other risk factors are excluded. Besides prolonged anti-NTM therapy, AOID-associated NTM infection should be treated with B-cell-depleting therapy to prevent recurrence.

Value of sputum Gram stain, sputum culture, and bronchoalveolar lavage fluid Gram stain in predicting single bacterial pathogen among children with community-acquired pneumonia

BackgroundCurrently, the microbial etiology of community-acquired pneumonia in children remains challenging. While Gram stain and sputum culture are commonly used to detect bacterial pathogens, it is unclear whether these approaches can predict single pathogen from bronchoalveolar lavage fluid (BALF) culture.MethodsA retrospective study involving 287 children hospitalized for pneumonia was conducted. Sputum specimens were collected on admission; and BALF specimens were collected within 24 h after admission. Taking BALF culture as the reference standard, the sensitivity and specificity of Sputum Gram stain (SGS), sputum culture, and BALF Gram stain (BGS) were calculated. The agreement between these approaches and BALF culture was compared using kappa statistics.ResultsFor SGS, the specificity was 23%. The overall sensitivity was 70%, including 87% for Gram-positive (G+) cocci, 56% for Gram-negative (G-) cocci, and 50% for G-bacilli. For sputum culture, the specificity was 70%. The overall sensitivity was 64%, including 71% for Streptococcus pneumoniae, 71% for Moraxella catarrhalis, and 64% for Haemophilus influenzae. For BGS, the specificity was 71%. The overall sensitivity was 60%, including 77% for G+cocci, 38% for G-cocci, and 44% for G-bacilli. While SGS had poor agreement with BALF culture, both sputum culture and BGS had moderate agreement with BALF culture.ConclusionsBoth sputum culture and BGS are helpful in predicting single bacterial pathogen from BALF culture among children with community-acquired pneumonia. Sputum cultures and BGS can provide early clues for BALF pathogen when BALF culture results are pending or bronchoscopy is not performed.

Protective effect of heat shock transcription factor 1 on acute lung injury in septic rats by regulating NOD-like receptor protein 3 inflammasome activation

To determine whether heat shock transcription factor 1 (HSF1) ameliorates sepsis induced acute lung injury (ALI) by regulating NOD-like receptor protein 3 (NLRP3) inflammasome activity in rat alveolar macrophages (AM). Twenty-four SPF Sprague-Dawley (SD) rats were divided into control group, lipopolysaccharide (LPS) group, overexpression empty vector+LPS group, and overexpression HSF1+LPS group by random number table method, with 6 rats in each group. The rat model of sepsis-induced ALI was reproduced by intraperitoneal injection of LPS (5 mg/kg); the rats in the control group were given the same volume of normal saline. The rats in the overexpression empty vector+LPS group and the overexpression HSF1+LPS group were instilled with 100 μL of overexpressed empty vector adenovirus or overexpressed HSF1 adenovirus through the trachea, respectively; the rats in the control group and LPS group were instilled with an equal volume of normal saline at the same time. At 6 hours after the model was reproduced, carotid blood was collected to determine the oxygenation index (PaO2/FiO2); lung tissue was obtained, and hematoxylin-eosin (HE) staining was performed to observe the pathological changes of lung tissue under a light microscope. Lung tissue wet/dry ratio (W/D) was determined. Immunohistochemistry was used to detect the positive expression of macrophage-specific marker antibody CD68. Western blotting was used to detect the protein expressions of HSF1 and NLRP3. Bronchoalveolar lavage fluid (BALF) was collected, and the levels of interleukins (IL-1β, IL-18) were detected by enzyme-linked immunosorbent assay (ELISA). In addition, BALF of normal rats was collected, and primary AM was isolated, cultured and divided into four groups. The AM in the blank control group was cultured normally without any treatment; the LPS group was treated with 1 mg/L LPS for 24 hours to reproduced the LPS stimulation model; AM in the overexpression empty vector+LPS group and the overexpression HSF1+LPS group were transfected with empty vector plasmid or overexpressed HSF1 plasmid, respectively, for 48 hours, and then the AM was treated with 1 mg/L LPS. The cell viability was detected by cell counting kit-8 (CCK-8). The protein expressions of HSF1 and NLRP3 in AM were detected by Western blotting. The levels of IL-1β and IL-18 in AM culture medium were determined by ELISA. Compared with the control group, the rat lung structure in the LPS group was severely injured, the alveolar cavity and pulmonary interstitium were congested and edema, the alveolar walls were significantly thickened and ruptured, accompanied by a large number of inflammatory cells infiltration. The lung W/D ratio increased, the infiltration degree of macrophages increased, PaO2/FiO2 decreased, HSF1 protein expression decreased and NLRP3 protein expression increased in lung tissue and AM, and IL-1β and IL-18 levels in BALF and AM culture medium increased. Compared with the LPS group, the degree of lung injury in the overexpression HSF1+LPS group was significantly improved, the lung W/D ratio was significantly reduced (4.76±0.16 vs. 6.93±0.33, P < 0.05), the macrophage infiltration was reduced, PaO2/FiO2 increased significantly [mmHg (1 mmHg ≈ 0.133 kPa): 397.62±19.46 vs. 280.12±37.42, P < 0.05], HSF1 protein expression was significantly up-regulated in lung tissue and AM (HSF1/GAPDH: 0.90±0.04 vs. 0.61±0.04 in lung tissue, 1.10±0.10 vs. 0.57±0.08 in AM, both P < 0.05), NLRP3 protein expression was significantly down-regulated (NLRP3/GAPDH: 0.75±0.14 vs. 1.05±0.11 in lung tissue, 0.81±0.09 vs. 1.14±0.17 in AM, both P < 0.05), and the contents of IL-1β and IL-18 in BALF and AM medium were significantly decreased [IL-1β (ng/L): 7.82±0.45 vs. 14.09±0.58 in BALF, 11.11±0.46 vs. 16.66±0.96 in AM; IL-18 (ng/L): 50.44±3.30 vs. 66.31±5.67 in BALF, 43.95±0.88 vs. 73.52±1.23 in AM, all P < 0.05]. There was no significant difference in the detection indicators between the overexpression empty vector+LPS group and the LPS group. HSF1 attenuates LPS-induced ALI in rats, and the mechanism may be related to the inhibition of NLRP3 inflammasome activation in AM.

Diagnostic accuracy of different bronchoscopic specimens in sputum Xpert MBT/RIF- negative pulmonary TB patients.

Tuberculosis (TB) control remains a critical public health problem worldwide. Rapid diagnosis and proper treatment are beneficial for the effective control of tuberculosis transmission. Diagnostic challenges arise when a patient has a clinical and radiological suspicion of tuberculosis but cannot produce sputum, sputum acid-fast bacilli, or Xpert Mycobacterium tuberculosis/rifampicin (Xpert MTB/RIF) is negative, resulting in suboptimal management. As a result, more invasive techniques must be used on these patients to establish the diagnosis. A retrospective study recruited 330 suspected pulmonary TB patients with negative sputum of Xpert MBT/RIF who underwent bronchoscopy between March 2018 and December 2021. The diagnostic yields of bronchoalveolar lavage fluid (acid-fast bacilli, Xpert MTB/RIF, and culture) and histopathologic examination (HPE) were calculated and compared to the final diagnosis and culture as a gold standard. Out of 330 suspected pulmonary TB patients, 181 had a final diagnosis of TB, and 149 had non-TB. The sensitivity of BALF (culture, Xpert, acid-fast bacilli) and trans-bronchial lung biopsy (HPE) was 80.7%, 72.9%, 21.1%, and 87.1%, respectively. Multiple nodules were associated with significantly higher BALF Xpert MTB, bronchoalveolar lavage fluid culture, and trans-bronchial lung biopsy (HPE) positivity. Bronchoscopic specimens are essential for accurate and rapid diagnosis of sputum Xpert MBT/RIF negative patients with high clinical and radiological suspicion of tuberculosis.

The lymphocyte-specific protein tyrosine kinase-specific inhibitor A-770041 attenuates lung fibrosis via the suppression of TGF-β production in regulatory T-cells.

Lymphocyte-specific protein tyrosine kinase (Lck) is a member of the Src family of tyrosine kinases. The significance of Lck inhibition in lung fibrosis has not yet been fully elucidated, even though lung fibrosis is commonly preceded by inflammation caused by infiltration of T-cells expressing Lck. In this study, we examined the effect of Lck inhibition in an experimental mouse model of lung fibrosis. We also evaluated the effect of Lck inhibition on the expression of TGF-β1, an inhibitory cytokine regulating the immune function, in regulatory T-cells (Tregs). Lung fibrosis was induced in mice by intratracheal administration of bleomycin. A-770041, a Lck-specific inhibitor, was administrated daily by gavage. Tregs were isolated from the lung using a CD4+CD25+ Regulatory T-cell Isolation Kit. The expression of Tgfb on Tregs was examined by flow cytometry and quantitative polymerase chain reaction. The concentration of TGF-β in bronchoalveolar lavage fluid (BALF) and cell culture supernatant from Tregs was quantified by an enzyme-linked immunosorbent assay. A-770041 inhibited the phosphorylation of Lck in murine lymphocytes to the same degree as nintedanib. A-770041 attenuated lung fibrosis in bleomycin-treated mice and reduced the concentration of TGF-β in BALF. A flow-cytometry analysis showed that A-770041 reduced the number of Tregs producing TGF-β1 in the lung. In isolated Tregs, Lck inhibition by A-770041 decreased the Tgfb mRNA level as well as the concentration of TGF-β in the supernatant. These results suggest that Lck inhibition attenuated lung fibrosis by suppressing TGF-β production in Tregs and support the role of Tregs in the pathogenesis of lung fibrosis.