Multiple Pathogen Infections Research Articles

Improvements in Blood Profiles of Canines Naturally Infected with Triple Blood Pathogens (Babesia vogeli, Ehrlichia canis, and Anaplasma platys) Subsequent to Doxycycline Monotherapy.

Multiple blood pathogen infections are increasingly found in many areas, particularly in tropical regions. This study aimed to investigate the effectiveness and safety of using doxycycline monotherapy to treat triple blood pathogen infection in sheltered dogs. A total of 375 sheltered dogs were screened for blood pathogen using polymerase chain reaction technique (PCR). There were 34 dogs with triple infection (Babesia vogeli, Ehrlichia canis, and Anaplasma platys), and most dogs displayed anemia and thrombocytopenia. These dogs were treated with doxycycline (10 mg/kg/day) orally for four weeks, and their blood profiles were monitored. Almost all pathogens were undetectable by PCR by day 14 and pathogens were all cleared by day 28. Most blood profiles significantly improved after 14 days. The improvement continued after 28 days. The red blood cell and platelet count (PLT) were increased in similar trends between mild-to-moderate and severe thrombocytopenia groups. The average PLT in the mild-to-moderate thrombocytopenia group raised to normal level by day 14, whereas in the severe group, it was gradually increased to normal level by day 70. We propose using doxycycline monotherapy for 28 days to eliminate all pathogens, and facilitate recovery and welfare in dogs suffering with triple blood pathogen infections.

Multiple pathogen infections accompanied with ocular pathologies in a patient with high-titer Anti-IFN-γ autoantibodies: a case report

The typical clinical characteristic of patients with anti-IFN-γ autoantibodies (AIGAs) is primarily associated with infection caused by intracellular pathogens. With continued research, additional clinical characteristics have been gradually uncovered. Here, we present a case of multiple pathogen infections accompanied by ocular pathologies in a patient with high titers of AIGAs. The patient, a 53-year-old female patient, was admitted to our hospital after finding a mass in the right supraclavicular fossa. She was successively diagnosed with Talaromyces marneffei, Aspergillus flavus and Nontuberculous mycobacteria (NTM) infections. Then, she received a complete course of antifungal agents for nearly 3.5 years and anti-NTM treatment for nearly 3 years, with discontinuation upon symptom improvement. However, there was a rapid recurrence of the infection upon cessation of the drug despite improvement in the patient’s symptoms. Moreover, when the recurrent infection stabilized, the patient exhibited immune conjunctivitis and dry eye, which was successfully treated by tacrolimus eye drops and lubricant. Patients with high-titer AIGAs are more prone to experiencing recurrence and/or persistent infection, as well as immune disorders.

Phase Transition of Wax Enabling CRISPR Diagnostics for Automatic At-Home Testing of Multiple Sexually Transmitted Infection Pathogens.

Sexually transmitted infections (STIs) significantly impact women's reproductive health. Rapid, sensitive, and affordable detection of these pathogens is essential, especially for home-based self-testing, which is crucial for individuals who prioritize privacy or live in areas with limited access to healthcare services. Herein, an automated diagnostic system called Wax-CRISPR has been designed specifically for at-home testing of multiple STIs. This system employs a unique strategy by using the solid-to-liquid phase transition of wax to sequentially isolate and mix recombinase polymerase amplification (RPA) and CRISPR assays in a microfluidic chip. By incorporating a home-built controlling system, Wax-CRISPR achieves true one-pot multiplexed detection. The system can simultaneously detect six common critical gynecological pathogens (CT, MG, UU, NG, HPV 16, and HPV 18) within 30min, with a detection limit reaching 10-18 M. Clinical evaluation demonstrates that the system achieves a sensitivity of 96.8% and a specificity of 97.3% across 100 clinical samples. Importantly, eight randomly recruited untrained operators performe a double-blinded test and successfully identified the STI targets in 33 clinical samples. This wax-transition-based one-pot CRISPR assay offers advantages such as low-cost, high-stability, and user-friendliness, making it a useful platform for at-home or field-based testing of multiple pathogen infections.

Low-cost high-throughput targeted sequencing for the accurate detection of respiratory tract pathogens.

The current gold standard for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis by real-time reverse transcriptase polymerase chain reaction (RT-PCR) is limited by the number of genes that can be detected. In this study, we developed a low-cost and high-throughput next-generation sequencing technology that can overcome the limitations of real time RT-PCR. A targeted sequencing panel (TSP) consisting of approximately 500 amplicons was designed. This panel could simultaneously detect a broad range of gene loci of SARS-CoV-2, and genes for the most common infectious viruses that affect the respiratory tract, in a single run and could include up to 96 samples. Four hundred and forty-eight samples and 31 control samples were analyzed independently with both TSP and RT-PCR, and the results were compared for accuracy and other indicators. TSP identified 50 SARS-CoV-2 positive samples with a 99.33% match to RT-PCR results. It is not surprising that TSP also identified multiple infections from the 96 samples, whereas RT-PCR could not. Thus, TSP was able to accurately diagnose the samples which could not be identified based on single RT-PCR test. Our data demonstrated that TSP is a fast and accurate testing method for identifying multiple pathogen infections of the respiratory tract.

CFTR Inhibitors Display Antiviral Activity against Herpes Simplex Virus.

The cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-dependent Cl- channel, is closely associated with multiple pathogen infections, such as SARS-CoV-2. However, whether the function of the CFTR is involved in herpes simplex virus (HSV) infection has not been reported. To evaluate the association of CFTR activity with HSV infection, the antiviral effect of CFTR inhibitors in epithelial cells and HSV-infected mice was tested in this study. The data showed that treatment with CFTR inhibitors in different concentrations, Glyh-101 (5-20 μM), CFTRi-172 (5-20 μM) and IOWH-032 (5-20 μM), or the gene silence of the CFTR could suppress herpes simplex virus 1 (HSV-1) and herpes simplex virus 2 (HSV-2) replication in human HaCaT keratinocytes cells, and that a CFTR inhibitor, Glyh-101 (10-20 μM), protected mice from HSV-1 and HSV-2 infection. Intracellular Cl- concentration ([Cl-]i) was decreased after HSV infection via the activation of adenylyl cyclase (AC)-cAMP signaling pathways. CFTR inhibitors (20 μM) increased the reduced [Cl-]i caused by HSV infection in host epithelial cells. Additionally, CFTR inhibitors reduced the activity and phosphorylation of SGK1 in infected cells and tissues (from the eye and vagina). Our study found that CFTR inhibitors can effectively suppress HSV-1 and HSV-2 infection, revealing a previously unknown role of CFTR inhibitors in HSV infection and suggesting new perspectives on the mechanisms governing HSV infection in host epithelial cells, as well as leading to potential novel treatments.

Multiplex PCR for bacterial, viral and protozoal pathogens in persistent diarrhoea or persistent abdominal pain in Côte d’Ivoire, Mali and Nepal

In contrast to acute diarrhoea, the aetiology of persistent digestive disorders (≥ 14 days) is poorly understood in low-resource settings and conventional diagnostic approaches lack accuracy. In this multi-country study, we compared multiplex real-time PCR for enteric bacterial, parasitic and viral pathogens in stool samples from symptomatic patients and matched asymptomatic controls in Côte d’Ivoire, Mali and Nepal. Among 1826 stool samples, the prevalence of most pathogens was highest in Mali, being up to threefold higher than in Côte d’Ivoire and up to tenfold higher than in Nepal. In all settings, the most prevalent bacteria were EAEC (13.0–39.9%) and Campylobacter spp. (3.9–35.3%). Giardia intestinalis was the predominant intestinal protozoon (2.9–20.5%), and adenovirus 40/41 was the most frequently observed viral pathogen (6.3–25.1%). Significantly different prevalences between symptomatic and asymptomatic individuals were observed for Campylobacter, EIEC and ETEC in the two African sites, and for norovirus in Nepal. Multiple species pathogen infection was common in Côte d’Ivoire and Mali, but rarely found in Nepal. We observed that molecular testing detected multiple enteric pathogens and showed low discriminatory accuracy to distinguish between symptomatic and asymptomatic individuals. Yet, multiplex PCR allowed for direct comparison between different countries and revealed considerable setting-specificity.

Detection and Characterization of Bacterial and Viral Acute Gastroenteritis among Outpatient Children under 5 Years Old in Guangzhou, China.

Acute gastroenteritis (AGE) in children can be attributed to a multitude of bacterial and viral pathogens. The objective of this study was to investigate the epidemiology of bacterial and viral AGE in children and to compare clinical characteristics between single and multiple enteric pathogen infections. A total of 456 stool samples were collected from outpatient children under 5 years old with AGE, which were subsequently analyzed for nine bacteria and three viruses using the Luminex xTAG® Gastrointestinal Pathogen Panel. The presence of at least one pathogen was detected in 260 cases (57.0%), with Salmonella being the predominant agent, followed by norovirus, Campylobacter, and rotavirus. A total of 69 cases (15.1%) exhibited positive results for two or more enteric pathogens. Although certain co-infections demonstrated significant differences in primary clinical features compared with mono-infections, no statistical variance was observed in terms of disease severity. In outpatient children from southern China, Salmonella emerged as the most prevalent causative agent of AGE, succeeded by norovirus and Campylobacter. This study underscores the burden posed by coinfections and highlights the clinical characteristics associated with AGE when accompanied by coinfections among children under 5 years old.

A Low-cost High-throughput Targeted Sequencing for the Accurate Detection of Respiratory Tract Pathogen

Introduction: The current gold standard for SARS-CoV-2 diagnosis by real-time RT-PCR has limitations of gene numbers that can be detected. In this study, we developed a low-cost and high-throughput next-generation sequencing technology that can overcome the limitations of RT-PCR. Methodology: A targeted sequencing panel (TSP) consisting of approximately 500 amplicons was designed that can simultaneously detect a broad range of gene loci of SARS-CoV-2 and genes for the most common viruses of respiratory infectious viruses in a single run of up to 96 samples. 448 samples and 31 control samples were examined independently with both TSP and RT-PCR, results were compared for accuracy and other indicators. Results: TSP identified 50 SARS-CoV-2 positive samples with a 99.33% match to RT-PCR results. It is not surprising that TSP also identified multiple viral infections from 96 samples, whereas RT-PCR could not. TSP demonstrated its ability to conclude diagnosis for those undecided from RT-PCR tests. Conclusion: Our data demonstrated that TSP is a fast and accurate test for detecting multiple pathogen infections of the respiratory tract.

Multiple intracellular pathogen infections with ocular pathologies associated with adult-onset immunodeficiency due to anti-interferon-γ autoantibodies: a case report

BackgroundAutoantibodies against interferon-γ (IFN-γ) can inhibit IFN-γ-dependent signal transducer and activator of transcription 1 phosphorylation and thus increase the risk of infection with intracellular pathogens, such as Talaromyces marneffei (TM), nontuberculous mycobacteria (NTMs), and Mycobacterium tuberculosis (TB). Here, we report a rare case of triple infection caused by TM, NTM, and TB in a human immunodeficiency virus–negative patient.Case presentationA middle-aged female was admitted to our hospital after experiencing recurrent rash, cough, and expectoration for 4 months. She was successively diagnosed with NTM, TM, and TB infections without conventional immunosuppression-associated factors. However, after effective anti-infective treatment, the patient was confirmed to have allergic conjunctivitis and was successfully treated with corticosteroids and immunosuppressants. The most conspicuous characteristics were recurrent infection and immune disorders.ConclusionsHigh-titer anti-IFN-γ autoantibodies are strongly associated with severe and disseminated infections, such as NTM, TM, and TB. It is characterized by persistently high degree of inflammation and high immunoglobin levels.

M5C RNA methylation: a potential mechanism for infectious Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disorder caused by a variety of factors, including age, genetic susceptibility, cardiovascular disease, traumatic brain injury, and environmental factors. The pathogenesis of AD is largely associated with the overproduction and accumulation of amyloid-β peptides and the hyperphosphorylation of tau protein in the brain. Recent studies have identified the presence of diverse pathogens, including viruses, bacteria, and parasites, in the tissues of AD patients, underscoring the critical role of central nervous system infections in inducing pathological changes associated with AD. Nevertheless, it remains unestablished about the specific mechanism by which infections lead to the occurrence of AD. As an important post-transcriptional RNA modification, RNA 5-methylcytosine (m5C) methylation regulates a wide range of biological processes, including RNA splicing, nuclear export, stability, and translation, therefore affecting cellular function. Moreover, it has been recently demonstrated that multiple pathogenic microbial infections are associated with the m5C methylation of the host. However, the role of m5C methylation in infectious AD is still uncertain. Therefore, this review discusses the mechanisms of pathogen-induced AD and summarizes research on the molecular mechanisms of m5C methylation in infectious AD, thereby providing new insight into exploring the mechanism underlying infectious AD.

Multiplex detection of bacterial pathogens by PCR/SERS assay.

Bacterial infections are a leading cause of death globally. The detection of DNA sequences correlated to the causative pathogen has become a vital tool in medical diagnostics. In practice, PCR-based assays for the simultaneous detection of multiple pathogens currently rely on probe-based quantitative strategies that require expensive equipment but have limited sensitivity or multiplexing capabilities. Hence, novel approaches to address the limitations of the current gold standard methods are still in high demand. In this study, we propose a simple multiplex PCR/SERS assay for the simultaneous detection of four bacterial pathogens, namely P. aeruginosa, S. aureus, S. epidermidis, and M. smegmatis. Wherein, specific primers for amplifying each target gDNA were applied, followed by applying SERS nanotags functionalized with complementary DNA probes and Raman reporters for specific identification of the target bacterial pathogens. The PCR/SERS assay showed high specificity and sensitivity for genotyping bacterial pathogen gDNA, whereby as few as 100 copies of the target gDNA could be detected. With high sensitivity and the convenience of standard PCR amplification, the proposed assay shows great potential for the sensitive detection of multiple pathogen infections to aid clinical decision-making.

Interspecific Host Variation and Biotic Interactions Drive Pathogen Community Assembly in Chinese Bumblebees.

Bumblebees have been considered one of the most important pollinators on the planet. However, recent reports of bumblebee decline have raised concern about a significant threat to ecosystem stability. Infectious diseases caused by multiple pathogen infections have been increasingly recognized as an important mechanism behind this decline worldwide. Understanding the determining factors that influence the assembly and composition of pathogen communities among bumblebees can provide important implications for predicting infectious disease dynamics and making effective conservation policies. Here, we study the relative importance of biotic interactions versus interspecific host resistance in shaping the pathogen community composition of bumblebees in China. We first conducted a comprehensive survey of 13 pathogens from 22 bumblebee species across China. We then applied joint species distribution modeling to assess the determinants of pathogen community composition and examine the presence and strength of pathogen-pathogen associations. We found that host species explained most of the variations in pathogen occurrences and composition, suggesting that host specificity was the most important variable in predicting pathogen occurrences and community composition in bumblebees. Moreover, we detected both positive and negative associations among pathogens, indicating the role of competition and facilitation among pathogens in determining pathogen community assembly. Our research demonstrates the power of a pluralistic framework integrating field survey of bumblebee pathogens with community ecology frameworks to understand the underlying mechanisms of pathogen community assembly.

SIMULTANEOUS PATHOGEN DETECTION OF SHRIMP VIRUSES ON CULTURED TIGER SHRIMPS (Penaeus monodon) IN INDONESIA

The multiple-pathogen infection causes severe economic impact to shrimp industry in Indonesia and worldwide due to mass mortality and multiple abnormalities of the survived infected shrimps. However, multiple-pathogen detection tools in shrimp diseases have not yet widely used. The purpose in this study was to develop and applied simultaneous detection system using multiplex polymerase chain reaction (PCR) assay from natural infections caused by white spot syndrome virus (WSSV), infectious hypodermal and haematopoietic necrosis virus (IHHNV) and monodon baculovirus (MBV) in Black tiger shrimp culture. To analyze multiple-pathogen infections in the shrimp, the study designed and used three pairs of specific primers targeting DNA virus from the shrimp diseases. All amplifications used a specific master mix for multiplex PCR assay and standardized extracted nucleic acid from the samples. This mPCR assay successfully amplified the DNA of three viruses in a single tube-run by multiplex PCR for each virus. Based on the results, the study confirms that multiple-pathogen infection contributes the highest mass mortality rather than from single infection by either WSSV, IHHNV or MBV. This study also confirms that the mPCR assay is a faster, cheaper, and efficient method to detect and subsequently prevent the spreading of multi-pathogen shrimp diseases.

Epstein–Barr virus downregulates the α7 nicotinic acetylcholine receptor of CD8+ T lymphocytes might associate with coronary artery lesions in Kawasaki disease patients

ObjectivesKawasaki disease (KD) is a systemic vasculitis that is caused by immunological dysregulation in children exposed to pathogens like Epstein–Barr virus (EBV). Myocardial ischemia or infarction due to coronary artery lesions (CALs) might be lethal. However, it is unclear how pathogens, immunomodulation, and CALs interact, particularly in KD patients co-infected with the most widespread virus, EBV. MethodsWe investigated pathogen carriage and fundamental clinical data in 281 KD patients. Immunological differences between CALs and non-CALs in KD patients under different conditions were analyzed. Then, the effect of infection by different pathogens on the immune response was excluded, and most EBV co-infected KD patients were included to assess the incidence of CALs, the level of immune modulation, and regulatory mechanisms in different EBV infection states. ResultsOur results showed multiple pathogenic infections occur in KD patients, with EBV being the most prevalent. The incidence of CALs in the EBV-DNA (+) acute infection group, EBV-DNA (−) acute infection group, and EBV latent infection group was 0 (0/6), 27.27% (3/11) and 41.67% (10/24), respectively. The two groups were younger and had increased IL-6 levels and B cells, decreasing CD8+ T cells than the EBV-DNA (+) acute infection group. Interestingly, the increased B cells were not associated with immunoglobulin release. Additionally, these patients down-regulated α7 nicotinic acetylcholine receptor (α7nAChR) and downstream molecule PI3K/AKT/mTOR while activating the NF-κB. ConclusionPatients with different EBV infection statuses exhibit different incidences of CALs. In acute EBV-DNA (−) infected and latent EBV-infected patients, the number of CD8+ T cells decreased and downregulated CD8+ T cells' α7nAChR and PI3K/AKT/mTOR, which may associate with CALs, while the expression of NF-κB and the pro-inflammatory factor IL-6 was upregulated by inhibiting the anti-inflammatory molecule α7nAChR.

Metagenomic next-generation sequencing versus traditional laboratory methods for the diagnosis of central nervous system opportunistic infections in HIV-infected Chinese adults

To evaluate clinical value of metagenomic next-generation sequencing (mNGS) in people living with HIV/AIDS (PLWHA) who had CNS disorders. Cerebrospinal fluid (CSF) samples from 48 PLWHA presenting with CNS disorders were sequenced using mNGS and compared with clinical conventional diagnostic methods. In total, 36/48 ss(75%) patients were diagnosed with pathogen(s) infection by mNGS, and the positive detection proportion by mNGS was higher than that by clinical conventional diagnostic methods (75% vs 52.1%, X2 = 5.441, P = 0.020). Thirteen out of 48 patients (27.1%) were detected with 3–7 pathogens by mNGS. Moreover, 77 pathogen strains were detected, of which 94.8% (73/77) by mNGS and 37.0% (30/77) by clinical conventional methods (X2 = 54.206, P < 0.001). The sensitivity and specificity of pathogens detection by mNGS were 63.9% (23/36) and 66.7% (8/12), respectively, which were superior to that by clinical conventional methods (23/36 vs 9/25, X2 = 4.601, P = 0.032; 8/12 vs 5/23, X2 = 5.029, P = 0.009). The application of mNGS was superior for its ability to detect a variety of unknown pathogens and multiple pathogens infection, and relatively higher sensitivity and specificity in diagnosis of CNS disorders in PLWHA.

Analysis of vaginal microecology in 23 181 cases of the gynecological female outpatients

Objective: To analyze the vaginal microecological status of vaginitis population and non-vaginitis population of gynecological female outpatients. Methods: A total of 30 265 women who visited the gynecological outpatient clinic of Beijing Obstetrics and Gynecology Hospital from December 2018 to December 2020 completed vaginal microecological examination. After removing the follow-up patients, 23 181 women were divided into group with symptoms and signs of vaginitis (6 697 cases) and group without symptoms and signs of vaginitis (16 484 cases), according to whether the women with symptoms and signs of vaginitis or not. And the vaginal microecological status of the two groups was compared and analyzed. Results: (1) The total detection rate of vaginitis in the initial women was 34.87% (8 083/23 181), of which 46.10% (3 087/6 697) in group with symptoms and signs of vaginitis and 30.31% (4 996/16 484) in group without symptoms and signs of vaginitis, nearly 1/3 of the gynecological outpatients without signs and symptoms of vaginitis had vaginitis. (2) Among the types of simple vaginitis, vulvovaginal candidiasis (VVC) was the most frequent in group with symptoms and signs of vaginitis (16.01%, 1 072/6 697), followed by aerobic vaginitis (AV; 12.83%, 859/6 697), with significant differences compared with group without symptoms and signs of vaginitis (all P<0.001). There were no statistical differences between the two groups of bacterial vaginosis (BV) and trichomonal vaginitis (TV), indicating that BV and TV were more likely to be neglected (all P>0.05). (3) The proportion of various combinations of vaginitis among 2 632 cases of mixed vaginitis were, in descending order: BV+AV, VVC+AV, BV+AV+VVC, AV+TV, AV+TV+BV, BV+VVC. (4) Microecological analysis of 15 098 cases diagnosed with non-vaginitis had normal flora (including those with normal flora and those with normal flora but decreased function) in 14 013 cases (92.81%, 14 013/15 098), abnormal flora in 429 cases (2.84%, 429/15 098) and the BV intermediate in 656 cases (4.34%, 656/15 098); this indicated that the vast majority of the microecological tests were normal in the vaginal microbiota of those without vaginitis. Conclusions: Microecological examination could diagnose multiple pathogenic infections at once, and is especially important as a guide for the definitive diagnosis of mixed vaginitis and vaginitis with atypical clinical symptoms. Vaginal infections such as BV and TV that are easily overlooked should be concerned.

Clinicopathologic Features and Pathogens of Granulomatous Lobular Mastitis

Background: Granulomatous lobular mastitis (GLM) is a rare, benign, and complex breast disease that can be easily misdiagnosed as breast cancer. The etiology of GLM is unclear, and optimal treatment has not been established. Methods: Medical records for 333 patients with GLM in recent 5 years at Longhua Hospital, Shanghai, China, were analyzed. Potential pathogens in 33 fresh tissue specimens were also analyzed using 16S rDNA sequencing technology, matrix-assisted laser desorption ionization time of flight mass spectrometry, and bacterial cultures. Results: The median age of patients was 32 years (range 22–47 years). Among 333 patients, 38.7% displayed elevated prolactin, while 23.7% displayed high interleukin-2. In the granulomatous lesion, CD3-positive T lymphocytes were significantly more than CD20-positive B lymphocytes around the vacuoles or microabscesses. Gram-positive organisms were observed in 82 cases, including in 22 cases from fresh tissue specimens. Thirty-three cases yielded associated pathogens and all displayed multiple pathogenic infections, as identified using 16S rDNA sequencing technology. Pathogenic infections were further identified as belonging to 16 main genera and 8 main pathogenic species. Conclusions: GLM displays distinct histological and clinical features similar to those that have been previously reported in the literature. Using 16S rDNA sequencing technology, all of our cases demonstrated multiple pathogenic infections, which provided more useful information for clinical treatment.

A Large Acute Gastroenteritis Outbreak Associated with Both Campylobacter coli and Human Sapovirus - Beijing Municipality, China, 2021.

Campylobacter is a significant foodborne pathogen that leads to global outbreaks of acute gastroenteritis (AGE) usually affecting less than 30 individuals. Human sapovirus (HuSaV) is an enteric virus responsible for sporadic cases and outbreaks of AGE worldwide. In a study conducted in Beijing, HuSaV detection ranked second after norovirus. We present a discussion of the first large-scale outbreak of AGE caused by both Campylobacter coli (C. coli) and HuSaV. The outbreak involved a total of 996 patients and exhibited two distinct peaks over a period of 17 days. Through case-control studies, we identified exposure to raw water from a secondary water supply system as a significant risk factor. Among 83 patients, 49 samples tested positive for C. coli, 39 samples tested positive for HuSaV, and 27 samples tested positive for both pathogens using real-time polymerase chain reaction detection. Furthermore, whole-genome sequencing of 17 C. coli isolates obtained from 17 patients revealed that all isolates belonged to a highly clonal strain of C. coli. Outbreaks of AGE resulting from multiple pathogen infections warrant increased attention. This report emphasizes the significance of ensuring the safety of drinking water, particularly in secondary supply systems.

Application of Alveolar Lavage Fluid Second-Generation Sequencing in the Treatment of Severe Pneumonia

To observe the application of alveolar lavage fluid second-generation sequencing in diagnosing and guiding the treatment of patients with severe pneumonia with unidentified pathogenic bacteria and to provide novel ideas and methods for the effective clinical treatment of this disease. The clinical data of 80 patients with severe pneumonia included in our intensive care unit from June 2020 to June 2022 were analyzed and all patients had undergone metagenomic next-generation sequencing and traditional cultures (alveolar lavage fluid culture, sputum culture) to analyze the advantages of metagenomic next-generation sequencing in detecting pathogens in patients with severe pneumonia. The positive rate of metagenomic next-generation sequencing pathogen detection was higher than that of conventional culture. In patients who were negative for conventional culture and positive for metagenomic next-generation sequencing, metagenomic nextgeneration sequencing was able to further identify multiple pathogenic infections. In terms of pathogen distribution, metagenomic next-generation sequencing detected 71 bacterial, 39 fungal and 3 viral strains. In mixed infections, metagenomic next-generation sequencing yielded a higher rate of positive diagnosis. In addition, metagenomic next-generation sequencing had a higher pathogen detection rate in patients with combined underlying diseases and metagenomic next-generation sequencing could identify specific pathogenic infections especially in patients with combined immunocompromised conditions. Metagenomic next-generation sequencing could improve the detection rate of pathogenic microorganisms in patients with severe pneumonia and could be used as a complementary test for patients negative for conventional cultures. Metagenomic next-generation sequencing has advantages in the diagnosis of mixed infections and can identify multiple pathogenic infections simultaneously. Clinically, early application of metagenomic next-generation sequencing in patients with severe pneumonia is recommended for increased clinical benefit.

Engineered Phage with Aggregation-Induced Emission Photosensitizer in Cocktail Therapy against Sepsis.

Sepsis, a widely recognized disease, is characterized by multiple pathogen infections. Therefore, it is imperative to develop methods that can efficiently identify and neutralize pathogen species. Phage cocktail therapy utilizes the host specificity of phages to adapt to infect resistant bacteria. However, its low sterilization stability efficiency and lack of imaging units seriously restrict its application. Here, a novel strategy combining the aggregation-induced emission photosensitizer (AIE-PS) TBTCP-PMB with phages through a nucleophilic substitution reaction between benzyl bromide and sulfhydryl groups to remove pathogenic bacteria for sepsis treatment is proposed. This strategy retains the phage's host specificity while possessing AIE-PS characteristics with a fluorescence imaging function and reactiveoxygenspecies (ROS) for detecting and sterilizing bacteria. This synergetic strategy combining phage cocktail therapy and photodynamic therapy (PDT) shows a strong "1+1>2" bactericidal efficacy and superior performance in sepsis mouse models with good biocompatibility. Furthermore, the strategy can quickly diagnose blood infections of clinical blood samples. This simple and accurate strategy provides a promising therapeutic platform for rapid pathogen detection and point-of-care diagnosis. Moreover, it presents a new method for expanding the library of antibacterial drugs to develop new strain identification and improve infectious disease treatment, thereby demonstrating strong translational potential.