Mixed Bacterial Infections Research Articles

The role of corneal biopsy in the management of microbial keratitis.

Corneal biopsy helps in diagnosing deep-seated or recalcitrant lesions of microbial keratitis (MK). We aim to analyze its role in managing these challenging cases. This is a retrospective review of 22 cases of corneal biopsy at our institute from January 2010 to December 2021. Data were retrospectively collected using the electronic medical record (EMR) system. Those cases of indolent, progressive MK or deep-seated lesions where cornea scraping was not possible were considered for corneal biopsy to establish the microbiological diagnosis. The primary aims of our study were to analyze the indications, success rates, and outcomes for biopsy patients in our series. Additional outcomes that were analyzed included the average time from presentation to biopsy, the type of causative organism isolated from the biopsy by either histopathological or microbiological method, and the frequency and outcome of surgical interventions performed. Descriptive statistics using mean (±standard deviation) and median (±range) were used to interpret the demographic data. Overall, 15 of 22 patients (68%) had a positive corneal biopsy after microbiological or histopathological examinations. The most identified organism was microsporidia (n = 4,30.7%), followed by mycobacteria (n = 2,15.4%), gram-negative bacilli (n = 2,15.4%), acid-fast bacilli (n = 1,7.6%), fungus (n = 2,15.4%), gram-positive cocci (n = 1,7.6%), and mixed bacterial infection (n = 1,7.6%). Corneal biopsy should be considered a diagnostic modality for patients with deep-seated or unresponsive MK. It can improve the treatment for MK, ensuring targeted therapy.

Dipicolylamine-Zn Induced Targeting and Photo-Eliminating of Pseudomonas aeruginosa and Drug-Resistance Gram-Positive Bacteria.

The emergence of drug-resistant bacteria, particularly resistant strains of Gram-negative bacteria, such as Pseudomonas aeruginosa, poses a significant threat to public health. Although antibacterial photodynamic therapy (APDT) is a promising strategy for combating drug-resistant bacteria, actively targeted photosensitizers (PSs) remain unknown. In this study, a PS based on dipicolylamine (DPA), known as WZK-DPA-Zn, is designed for the selective identification of P. aeruginosa and drug-resistant Gram-positive bacteria. WZK-DPA-Zn exploits the synergistic effects of DPA-Zn2+ coordination and cellular uptake, which could effectively anchor P. aeruginosa within a brief period (10min) without interference from other Gram-negative bacteria. Simultaneously, the cationic nature of WZK-DPA-Zn enhances its interaction with Gram-positive bacteria via electrostatic forces. Compared to traditional clinical antibiotics, WZK-DPA-Zn shows exceptional antibacterial activity without inducing drug resistance. This effectiveness is achieved using the APDT strategy when irradiated with white light or sunlight. The combination of WZK-DPA-Zn with Pluronic-based thermosensitive hydrogel dressings (WZK-DPA-Zn@Gel) effectively eliminates mixed bacterial infections and accelerates wound healing, thereby achieving a synergistic effect where "1+1>2." In summary, this study proposes a precise strategy employing DPA-Zn as the targeting moiety of a PS, facilitating the rapid elimination of P. aeruginosa and drug-resistant Gram-positive bacteria using APDT.

Heterogeneous Cu2O-SnO2 doped polydopamine fenton-like nanoenzymes for synergetic photothermal-chemodynamic antibacterial application

Wound infections caused by drug-resistant bacteria pose a great threat to human health, and the development of non-drug-resistant antibacterial approaches has become a research priority. In this study, we developed Cu2O-SnO2 doped polydopamine (CSPDA) triple cubic antibacterial nanoenzymes with high photothermal conversion efficiency and good Fenton-like catalase performance. CSPDA antibacterial nanoplatform can catalyze the generation of hydroxyl radical (·OH) from H2O2 at low concentration (50 μg∙mL−1) under 808 nm near-infrared (NIR) irradiation to achieve a combined photothermal therapy (PTT) and chemodynamic therapy (CDT). And the CSPDA antibacterial nanoplatform displays broad-spectrum and long-lasting antibacterial effects against both Gram-negative Escherichia coli (100 %) and Gram-positive Staphylococcus aureus (100 %) in vitro. Moreover, in a mouse wound model with mixed bacterial infection, the nanoplatform demonstrates a significant in vivo bactericidal effect while remaining good cytocompatible. To conclude, this study successfully develops an efficient and long-lasting bacterial infection treatment system. This system provided different options for future studies on the design of synergistic antimicrobial therapy. Hence, the as-synthesized synergetic photothermal therapy and chemodynamic therapy nanoenzymes have rapid and long-term bactericidal ability, well-conglutinant performance and effectively preventing wound infection for clinical application. Statement of SignificanceWound infections caused by drug-resistant bacteria pose a great threat to human health, and the development of non-drug-resistant antibacterial approaches has become a research priority. In this study, we developed Cu2O-SnO2 doped polydopamine (CSPDA) triple cubic yolk-like antibacterial nanoenzymes with high photothermal conversion efficiency and Fenton-like catalase effect for photothermal and Chemodynamic antibacterial therapy, Meanwhile, the nanocomposites exhibit good antibioadhesion in a natural water environment for a long-time immersion. In conclusion, this study successfully develops an efficient and long-lasting bacterial infection treatment system. These findings present a pioneering strategy for future research on the design of synergistic antibacterial and antibioadhesive systems.

Risk factors for catheter-related bloodstream infections in patients with intestinal failure undergoing home parenteral nutrition: a single-center study.

The incidence and risk factors of catheter-related bloodstream infections (CRBSI) in patients with intestinal failure (IF) have not been established, partly because catheter management methods vary from different facilities. This study aimed to identify the risk factors and incidence rate of CRBSIs in patients with IF who were given prophylactic treatment. Sixteen patients with IF who required home parenteral nutrition were enrolled in this study. Prophylactic management of CRBSI included monthly ethanol lock therapy and standardized infection prevention education. The outcomes included the incidence and risk factors of CRBSI. The median incidence rate of CRBSI was 1.2 per 1000 catheter days. Univariate analysis showed that the risk of developing CRBSI was significantly associated with short bowel syndrome (< 30cm) (p = 0.016). Other relevant findings included a significant negative correlation between serum albumin and CRBSI rate (r = -0.505, p = 0.046), and past history of mixed bacterial infections was significantly associated with increased CRBSI rate (p = 0.013). CRBSIs can still develop despite undergoing prophylactic management. Risk factors for CRBSI include the residual intestinal length, nutritional status, and susceptibility to certain microorganisms.

Bacterial Stomatitis in Wild Reticulated Pythons (Malayopython reticulatus) in Malaysia

Bacterial stomatitis is a common clinical form of upper alimentary tract disease in reptiles. The current study aimed to isolate and identify the common aerobes in the oral cavities of wild reticulated pythons and to profile their antimicrobial susceptibility. The need to conduct the current research was deemed in parallel with the increasing demand for snakes as pets and the growing emergence of multiple-drug-resistant organisms. A total of 40 fresh carcasses of the wild-caught reticulated pythons were assessed for the presence or absence of stomatitis. Oral swabs were obtained and cultured on blood and MacConkey agar media. The colony and cellular morphologies of the isolates were evaluated, followed by Gram-positive and Gram-negative bacterial identification. Antimicrobial susceptibility testing was performed using Kirby-Bauer disk diffusion method against selected antibiotics, namely gentamicin (GEN), amoxicillin (AMX), cephalexin (LEX), azithromycin (AZM), tetracycline (TET), and ciprofloxacin (CIP), commonly used to treat bacterial infection in reptiles. Results indicated that the prevalence of stomatitis was 77.5%. Among 153 isolates identified, 76.47% of bacteria were identified from pythons with stomatitis lesions, while 23.53% of bacteria were identified from pythons without stomatitis. Of 153 isolates, Gram-negative bacteria were shown to be predominant (94.77%). The three most isolated bacterial species were Aeromonas spp. (14.38%), Klebsiella pneumoniae (11.76%), and Alcaligenes faecalis (8.5%). Meanwhile, coagulase-negative Staphylococcus spp. (4.58%) and Corynebacterium spp. (0.66%) were the only isolated Gram-positive aerobes. Most isolates were observed to be equally susceptible to GEN and CIP (at 95.8%) but highly resistant to AMX (83.3%) and LEX (75.0%). In conclusion, bacterial stomatitis in wild-caught reticulated pythons was highly prevalent and often seen as a mixed bacterial infection (96.8%). The isolated bacteria consistently show susceptibility towards GEN and CIP and thus could be considered the primary line of antibiotics in treating this disease.

Clinical characteristics and mortality risk factors of mixed bacterial infections in hematopoietic stem cell transplantation recipients.

Mixed bacterial infections (MBI) is one of the complications after hematopoietic stem cell transplantation (HSCT) and increases the risk of patient death. However, there are few reports specifically on this topic. The purpose of this study was to investigate the clinical characteristics and mortality risk factors of MBI in HSCT recipients. The electronic medical records of patients undergoing HSCT were collected. The epidemiological features and antibiotic resistance of patients with and without MBI were compared. Logistic regression and Cox regression were used to identify the risk factors for MBI acquisition and death. R language was used to construct a prediction model for the overall survival of HSCT recipients with MBI. The cumulative incidence of MBI was 6.3% and the mortality was 48.8%. Time interval from diagnosis to transplantation > 180 days (HR=2.059, 95% CI 1.042-4.069, P=0.038) and ICU admission after transplantation (HR=2.271, 95% CI 1.053-4.898, P=0.036) were independent risk factors for MBI acquisition. Engraftment period > 20 days (HR=2.273, 95% CI 1.028-5.027, P=0.043), continuous renal replacement therapy (HR=5.755, 95% CI 1.691-19.589, P=0.005) and septic shock (HR=4.308, 95% CI 2.085-8.901, P=0.000) were independent risk factors associated with mortality. MBI has become a serious problem that cannot be ignored after HSCT. It is urgent for clinicians to pay high attention to it and formulate reasonable monitoring and treatment plans to improve the prognosis of patients.

Secondary Infection Surveillance with Metagenomic Next-Generation Sequencing in COVID-19 Patients: A Cross-Sectional Study.

Metagenomic next-generation sequencing (mNGS) is a promising tool for improving antimicrobial therapy and infection control decision-making in complex infections. Secondary infection surveillance using mNGS in COVID-19 patients has rarely been reported. Respiratory pathogen and antibiotic resistance prediction were evaluated by BALF mNGS for 192 hospitalized COVID-19 patients between December 2022 and February 2023. Secondary infection was confirmed in 83.3% (160/192) of the COVID-19 patients, with bacterial infections (45%, 72/160) predominating, followed by mixed bacterial and fungal infections (20%, 32/160), and fungal infections (17.5%, 28/160). The incidence of bacterial or viral secondary infection was significantly higher in patients who were admitted to the ICU, received mechanical ventilation, or developed severe pneumonia (all p<0.05). Klebsiella pneumoniae (n=30, 8.4%) was the most prevalent pathogen associated with secondary infection followed by Acinetobacter baumannii (n=29, 8.1%), Candida albicans (n=29, 8.1%), Aspergillus fumigatus (n=27, 7.6%), human herpes simplex virus type 1 (n=23, 6.4%), Staphylococcus aureus (n=20, 5.6%) and Pneumocystis jiroveci (n=14, 3.9%). The overall concordance between the resistance genes detected by mNGS and the reported phenotypic resistance in 69 samples containing five clinically important pathogens (ie, K. pneumoniae, A. baumannii, S. aureus, P. aeruginosa and E. coli) that caused secondary infection was 85.5% (59/69). mNGS can detect pathogens causing secondary infection and predict antimicrobial resistance for COVID19 patients. This is crucial for initiating targeted treatment and rapidly detect unsuspected spread of multidrug-resistant pathogens.

Modern views on the correction of violations of the vaginal biotope in perimenopause

ABSTRACT. Ensuring the quality of life of women in the climacteric period remains today the most urgent problem of modern medicine. The goal is to evaluate the effectiveness of the antiseptic Ginodec in the treatment of mixed vulvovaginal infection in perimenopausal women. Materials and methods. We monitored 52 patients with mixed bacterial and candidal infection aged 44 to 52 years. The diagnosis of non-specific bacterial and candidal infection was verified according to the data of clinical and laboratory research methods. The criteria for the effectiveness of the treatment were complete clinical and bacterial sanitation. Patients with bacterial candidal infection received 5 ml of Ginodek vaginal gel once a night for 7 days. The results. Analysis of clinical effectiveness after treatment showed that in perimenopausal women, complaints of vaginal discomfort and vaginal discharge decreased by 4 times, dyspareunia was noted by only 5.8 % of patients compared to 69.2 % before treatment. Regenerative efficiency of vaginal epithelial cells in patients also improved, their maturity index increased to 75-80. Bacteriological examination carried out 2 weeks after the end of the course of treatment confirmed the effectiveness of the Ginodek drug against bacterial infection in 92.3 % of cases, candidal - in 86.5 %. Conclusions. Bacterial-fungal associations of microorganisms in the vaginal biotope in women in the perimenopausal period lead to the formation of a complex complex of unclear clinical symptoms, which makes it difficult to make a timely diagnosis. The drug Ginodek demonstrates high microbiological effectiveness in mixed bacterial-candidal infection in patients in perimenopause, and also increases the functional activity of the epithelium of the mucous membrane of the vagina, which is especially important in this period of a woman's life.

The microbiological diagnostic performance of metagenomic next-generation sequencing in patients with infectious diseases.

Metagenomic next-generation sequencing (mNGS) can be used to detect pathogens in clinical infectious diseases through the sequencing analysis of microbial and host nucleic acids in clinical samples. This study aimed to assess the diagnostic performance of mNGS in patients with infections. In this study, 641 patients with infectious diseases were enrolled. These patients simultaneously underwent pathogen detection by both mNGS and microbial culture. Through statistical analysis, we judged the diagnostic performance of mNGS and microbial culture on different pathogens. Among 641 patients, 276 cases of bacteria and 95 cases of fungi were detected by mNGS, whereas 108 cases of bacteria and 41 cases of fungi were detected by traditional cultures. Among all mixed infections, combined bacterial and viral infections were the highest (51%, 87/169), followed by combined bacterial with fungal infections (16.57%, 28/169) and mixed bacterial, fungal, and viral infections (13.61%, 23/169). Among all sample types, bronchoalveolar lavage fluid (BALF) samples had the highest positive rate (87.8%, 144/164), followed by sputum (85.4%, 76/89) and blood samples (61.2%, 158/258). For the culture method, sputum samples had the highest positive rate (47.2%, 42/89), followed by BALF (37.2%, 61/164). The positive rate of mNGS was 69.89% (448/641), which was significantly higher than that of traditional cultures (22.31% [143/641]) (P < .05). Our results show that mNGS is an effective tool for the rapid diagnosis of infectious diseases. Compared with traditional detection methods, mNGS also showed obvious advantages in mixed infections and infections with uncommon pathogens.

Clinical characteristics of intestinal mixed infections

Objective. To study the features of the clinical picture of mixed intestinal infections in patients treated at the Regional Infectious Clinical Hospital named after A.M. Nichoga in the period from 2017 to 2021.&#x0D; Materials and methods. The clinical picture was studied in 55 patients with mixed intestinal infection who were treated at the Regional Infectious Hospital named after A.M. Nichoga (Astrakhan).&#x0D; Results. Mixed intestinal infections in most cases were registered in the summer-autumn period (51 % 92.7 %) in the form of sporadic morbidity. Epidemiological anamnesis data show that the majority of patients drank unboiled water, swam in the river, where they accidentally swallowed water, consumed food products of questionable quality, chicken meat, eggs insufficiently heat-treated, unwashed vegetables and fruit. Four patients arrived in Astrakhan from amoebiasis-endemic regions (Uzbekistan, Turkmenistan, Turkey, Egypt); 51 patients (92.7 %) had a moderate form of the disease,4 patients (7.3 %) a severe form. The clinical picture of mixed intestinal inflections was characterized by the symptoms of shigellosis and salmonellosis.&#x0D; Two patients, diagnosed shigellosis + amoebiasis had manifestations of ulcerative colitis complicated by intestinal bleeding; 2 patients, diagnosed salmonellosis + amoebiasis, developed dehydration shock. The treatment of patients with a severe form was carried out in the Intensive Care Unit and Resuscitation Unit of the Regional Infectious Clinical Hospital.&#x0D; Conclusions. The clinical picture of mixed bacterial intestinal infections and intestinal amoebiasis is mainly determined by the symptoms of shigellosis and salmonellosis and is manifested by the phenomena of intoxication, dehydration and the development of colitic and gastroenterocolitic syndromes.

Mixed bacterial-viral lung infections in patients with cystic fibrosis

The main cause of death in patients with cystic fibrosis (CF) is infectious process in the lungs, in particular, chronic lung infections caused by various pathogens, most often a combination of bacteria, fungi, or viruses. Data on mixed bacterial and viral-bacterial infections from domestic and foreign sources are fragmentary and sparse. The dominant associations of bacterial and viral pathogens in patients with cystic fibrosis have not been studied properly, and data on their epidemiological significance are lacking. The aim of this study was to assess the prevalence of bacterial and viral infections in patients with cystic fibrosis and to substantiate the need for the development of virological monitoring. Methods. Biomaterials from the respiratory tract of CF patients (409 children and 160 adults with CF) examined from 2006 to 2022 were used. The study was carried out using bacteriological methods, molecular genetic methods (RT-PCR) and MALDI-TOF mass-spectrometry. Results. Microflora of the respiratory tract was shown to be mixed in 2/3 patients with CF. The microflora of the lungs of children with CF is a dynamic community of microorganisms with high diversity and variability. In adult patients, associations of microorganisms are more common than in children, but the composition of associations is less diverse. We isolated about 40 species of bacteria from adult patients and more than 85 species from children in our sample. NFMO prevailed, including Burkholderia cepacia complex, Pseudomonas aeruginosa, Achromobacter xylosoxidans, Achromobacter ruhlandii, Stenotrophomonas maltophilia, and Staphylococcus aureus, Candida albicans, Aspergillus spp. Real-time PCR showed the presence of rhinovirus RNA in 10% of samples obtained from children and 12.9% from adults with cystic fibrosis. Conclusion. Our results indicate the need for continuous monitoring of the lung microflora in patients with CF, including testing for viruses.

Epidemiologic, clinical, and serum markers may improve discrimination between bacterial and viral etiologies of childhood pneumonia.

Discrimination of bacterial and viral etiologies of childhood community-acquired pneumonia (CAP) is often challenging. Unnecessary antibiotic administration exposes patients to undue risks and may engender antimicrobial resistance. This study aimed to develop a prediction model using epidemiological, clinical and laboratory data to differentiate between bacterial and viral CAP. Data from 155 children with confirmed bacterial or mixed bacterial and viral infection (N = 124) and viral infection (N = 31) were derived from a comprehensive assessment of causative pathogens [Partnerships for Enhanced Engagement in Research-Pneumonia in Pediatrics (PEER-PePPeS)] conducted in Indonesia. Epidemiologic, clinical and biomarker profiles (hematology and inflammatory markers) were compared between groups. The area under the receiver operating characteristic curve (AUROC) for varying biomarker levels was used to characterize performance and determine cut-off values for discrimination of bacterial and mixed CAP versus viral CAP. Diagnostic predictors of bacterial and mixed CAP were assessed by multivariate logistic regression. Diarrhea was more frequently reported in bacterial and mixed CAP, while viral infections more frequently occurred during Indonesia's rainy season. White blood cell counts (WBC), absolute neutrophil counts (ANC), neutrophil-lymphocyte ratio (NLR), C-reactive protein (CRP), and procalcitonin (PCT) were significantly higher in bacterial and mixed cases. After adjusting for covariates, the following were the most important predictors of bacterial or mixed CAP: rainy season (aOR 0.26; 95% CI 0.08-0.90; p = 0.033), CRP ≥5.70 mg/L (aOR 4.71; 95% CI 1.18-18.74; p = 0.028), and presence of fever (aOR 5.26; 95% CI 1.07-25.91; p = 0.041). The model assessed had a low R-squared (Nagelkerke R2 = 0.490) but good calibration (p = 0.610 for Hosmer Lemeshow test). The combination of CRP and fever had moderate predictive value with sensitivity and specificity of 62.28 and 65.52%, respectively. Combining clinical and laboratory profiles is potentially valuable for discriminating bacterial and mixed from viral pediatric CAP and may guide antibiotic use. Further studies with a larger sample size should be performed to validate this model.

An end-point multiplex PCR/reverse transcription-PCR for detection of five agents of bovine neonatal diarrhea

Neonatal calf diarrhea (NCD) is frequently associated with single or mixed viral, bacterial and/or protozoal infections. Consequently, laboratory diagnostic of NCD usually requires specific tests for each potential agent; a time-consuming, laborious and expensive process. Herein, we describe an end-point multiplex PCR/reverse transcription-PCR (RT-PCR) for detection of five major NCD agents: bovine rotavirus (BRV), bovine coronavirus (BCoV), Escherichia coli K99 (E. coli K99), Salmonella enterica (S. enterica) and Cryptosporidium parvum (C. parvum). Initially, we selected and/or designed high-coverage primers. Subsequently, we optimized multiplex PCR/RT-PCR conditions. Next, we evaluated the analytical sensitivity of the assay and assessed the performance of the reaction by testing 95 samples of diarrheic calf feces. The analytical specificity was evaluated against bovine viral diarrhea virus (BVDV), E. coli heat-stable enterotoxin (STa) and Eimeria spp. The detection limit of our assay was about 10 infectious units of BRV, 10−2 dilution of a BCoV positive sample pool, about 5 × 10−4 CFU for S. enterica, 5 × 10−6 CFU for E. coli K99 and 50 oocysts for C. parvum. No non-specific amplification of other bovine diarrhea agents was detected. Out of 95 samples analyzed, 50 were positive for at least one target, being 35 single and 15 mixed infections. BRV was the most frequent agent detected in single infections (16/35), followed by Cryptosporidium spp. (11/35), which was the most frequent in mixed infections (11/15). Positive and negative multiplex results were confirmed in individual reactions. In conclusion, we described an end-point multiplex PCR/RT-PCR for faster and easier NCD diagnosis, which may be useful for routine diagnosis and surveillance studies.

PREVALENCE OF CAUSATIVE AGENTS OF RING ROT CLAVIBACTER SEPEDONICUS, BLACKLEG AND WET ROT PECTOBACTERIUM ATROCEPTICUM IN THE 2021 YEAR POTATO HARVEST IN THE TERRITORY OF UKRAINE

Aim. To investigate the prevalence of causative agents of ring rot and soft rot in the potato harvest by the DAS ELISA method. Materials and methods. Samples of the potato harvest from Ternopil, Zhytomyr, Kyiv, Cherkasy, Dnipropetrovsk, Donetsk, Mykolaiv, Odesa, and Kherson regions were used in the study. The detection of bacterial agents was conducted by enzyme-linked immunosorbent assay using commercial test systems LOEWE® Standard Complete Kit (Germany). Results. The highest percentage of affected potatoes was registered in the Donetsk region: 11.1% – by the Clavibacter seedonicus, 31.0% – by the Pectobacterium atrosepticum, and 8.9% – by both bacteria. The causative agent of soft rot was detected twice as often as a causative agent of ring rot in tuber samples from all regions, except the Ternopil region, where the predominance of Clavibacter sepedonicus was revealed. In addition to the Donetsk region, a high prevalence of the causative agent of soft rot was found in the potato harvest from Kyiv (10.2%) and Odesa (10.0%) regions, the highest abundance of the causative agent of ring rot was detected in the Ternopil region. The mixed bacterial infection of potato tubers was diagnosed in 6 areas, and the highest rate was detected in the Donetsk region. Conclusion. The analysis of the distribution of ring rot pathogen Clavibacter sepedonicus and soft rot pathogen Pectobacterium atrocepticum in the potato crop shows the presence of research pathogens in all studied regions. It was discovered the dominance of the representative of pectolytic bacteria in all regions except Ternopil, where the distribution of Clavibacter sepedonicus, the causative agent of ring rot, was dominated. High rates of spread of the studied pathogens were associated with increased local average annual precipitation, as well as with increased annual amplitude of this indicator.

Comparison of intestinal bacterial communities in asymptomatic and diseased Asian seabass (Lates calcarifer) with chronic enteritis and mixed bacterial infections

Asian seabass (Lates calcarifer) is a major aquaculture food fish species in Singapore. Farming of this species is increasingly threatened by frequent outbreaks of infectious diseases, resulting in mortality exceeding 50–70%. In this study, we investigated the comparative gut bacterial microbiota using 16S rRNA metasequencing between asymptomatic and diseased juvenile fish collected during a disease outbreak soon after stocking. Mild to severe chronic granulomatous enteritis was observed histopathologically in both asymptomatic and diseased fish. Kidneys of diseased fish tested PCR positive for the ‘big belly’ novel Vibrio spp., Streptococcus iniae and Vibrio harveyi. These bacteria were also readily detected by PCR in water samples corresponding to tanks fish were sampled from. Potentially beneficial microbes that promote gut health such as Firmicutes, Bacteroidota and Actinobacteriota were the dominant phyla in the intestinal microbiota of asymptomatic fish. Moreover, the bacteria with probiotic potential such as Lactobacillus only presented in asymptomatic fish, and Weissella was unique and prevalent (47.59%) in asymptomatic fish during the recovery phase of the disease outbreak, making them candidate biomarkers for monitoring health status of L. calcarifer. Conversely, diseased fish showed reduced diversity of their gut microbiome, with high abundance of members of the phylum Proteobacteria. Vibrio was the most dominant genus (87.3%) and Streptococcus iniae was only detected in diseased fish. These findings provide a baseline study for understanding changes in intestinal microbiota in newly stocked fish with mixed bacterial infection, biomarker assisted health monitoring, and future host-derived probiotics screening in L. calcarifer.

The Role of IL-6 and IL-17 in SARS CoV-2 Patients with Secondary Bacterial Pneumonia

The current study was conducted in Kirkuk city at Al-Shifaa 14 hospital from November 2021 to March 2022, indicated the bacteria causing secondary pneumonia isolated from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) patients and the role of IL-6 and IL-17 in these infection.&#x0D; Sputum samples were used to obtain the bacterial isolates, and API testing was used to confirm the species level identification. Using an enzyme-linked immunosorbent assay (sandwich ELISA), the levels of IL-6 and IL-17 in the blood were evaluated. The study documented several bacterial species in a single infection (56/87.5%) or mixed bacterial infection (8/12.5%). The most common isolated bacteria species was Klebsiella pneumoniae (35.95%) followed by Staphylococcus aureus (31.25%), E. coli (17.19%), Pseudomonas aeruginosa (10.94%), and (1.56%) for each of Klebsiella oxytoca, Acinetobacter baumannii and Cronobacter sakazakii. The study recorded a high significant difference (P &lt;0.01) between the patients (22.2±6.82) pg/mL and the control group (58.39±11.15) pg/ mL concerning IL-6 also a high significant difference (P &lt;0.01) between the patients (101.79±27.13) pg/mL and the control group (58.39±11.15) pg/mL concerning IL-17.&#x0D; In conclusion, K. pneumoniae and S. aureus were the predominant isolated bacteria from COVID-19 patient's lung and there was a highly significant increase in IL-6 and IL-17 levels in secondary bacterial pneumonia in COVID-19 patients.

Incidence of symptomatic aerobic vaginitis among some Iraqi women in Baghdad city

Aerobic vaginitis (AV) is a newly adopted type of vaginal infection caused by aerobic bacteria; it was defined by Donders in 2002 and diagnosed according to Donder's criteria. AV is associated with increased vaginal pH, decreased vaginal microbiota and overgrowth of facultative anaerobic or aerobic bacteria, including Gram-positive cocci and Gram-negative bacilli. Because knowledge of the aerobic bacterial types causing AV is very important and only limited studies are available in Iraq about this type of vaginal infection, this study aimed to report the prevalence of AV in symptomatic women in Baghdad City, investigate the aerobic bacterial types associated and to evaluate the most critical symptoms and risk factors associated with AV. One hundred fifteen high vaginal swabs (HVSs) and 115 vaginal swabs (VSs) were collected from women of age 18-50 years attending some hospitals and private clinics in Baghdad City under the supervision of a competent gynecologist. Vaginal swabs were prepared for direct wet mount preparation to test under the microscope and Gram staining for AV identification according to Donders Criteria. High vaginal swabs were cultured on different culture media; the primary diagnosis of obtained colonies was based on phenotypic characteristics, conventional biochemical tests and Gram staining. The diagnosis was confirmed using Gram-positive (GP) and Gram-negative (GN) identification cards of the VITEK 2 System. Statistical analysis was carried out using (SPSS v 20) and p-value ≤ 0.05 at 95% CI was considered statistically significant. Out of the 115 swabs, only 89 (77.3%) swabs showed bacterial growth. Ninety-five bacterial isolates were obtained, including (65.2%) Gram-positive and (34.7%) Gram-negative bacteria. G+ve bacterial isolates included: (46.3%) Staphylococcus spp., (6.3%) Kocuria spp., (6.3%) Enterococcus spp., (5.2%) Micrococcus luteus and (1.0%) of Streptococcus agalactiae. G-ve bacterial isolates included (15.7%) Escherichia coli, (11.2%) Klebsiella pneumoniae, (3.2%) Pseudomonas aeruginosa (3.2%) Pseudomonas aeruginosa, (2.1%) Acinetobacter baumannii and (2.1%) Proteus mirabilis. More than half (58.4%) of patients showed severe AV. Mixed bacterial infections were reported in 6 (6.7%) cases only. Vaginal pH ranged between (5.5- 6.5). The most frequent(100%) testified symptoms were abnormal vaginal discharge and itching, or irritation, the less frequent symptoms (51.6%) was vaginal dyspareunia, and the most significant symptoms in associated with different bacterial types and age groups were foul smelling and burning (p= 0.001, 0.008, 0.0001). Among the risk factors, age was significantly associated with most bacterial types obtained with p values (0.05 and 0.02). Staphylococcus spp. and E.coli were the predominant bacterial types in AV patients in the current study, and the lower rate of bacteria was Streptococcus agalactiae (1.0%). A high prevalence of AV was reported; therefore, regular screening and proper diagnosis of AV using microscopic examinations, culturing of swabs and determining vaginal pH using specific vaginal pH test strips should be stimulated to develop AV management. Keywords: High vaginal swabs, Vaginal swabs, Aerobic vaginitis, aerobic G+ve and G-ve bacteria, Risk factors, Iraq.

THE COMPARISON OF MACERATION AND SOXHACLETATION METHODS EFFECTIVENESS ON GARLIC EXTRACT TO INHIBIT STAPHYLOCOCUS AUREUS GROWTH

Introduction: An abscess is pathological, pus-filled cavity caused by mixedbacterial infection. Bacteria that a role in the process, namely Staphyloccocusaureus. Treatment of abscesses is usually by antibiotics. Abscesses caused byStaphylococcus aureus appears to be resistant to antibiotics, alternative needed by utilizing herbal plants such as garlic. Garlic has active ingredientallicin has antibacterial. Objective: The purpose was to determined whichmethod was the most effective among maceration and soxhletation methodsin making garlic extract with a concentration of 50% against the resistance ofStaphyloccocus aureus. Material and Methods: The research method usedKirby Bauer method which is sensitivity test with the agar diffusion methodusing the disc diffusion technique. The Kirby Bauer sensitivity test usingselective media, namely Muller Hinton Agar media, the inhibition zone iscreated area with a clear zone around the disc. Result: Data obtained fromthe results this study were statistically tested using normality with Shapiro-Wilk to see whether the data normally distributed or not, the expression was with the One Way Anova test to see whether there was a difference that could be measured later with the Least Significant Differencetest to see groups that are influential in inhibiting bacteria. Discussion: Thecontent of active compounds from garlic extract by soxhletation methodmore easily extracted because circulation that can increase the rate of transfercompounds from extract. Conclusion: The results, 50% concentration ofgarlic extract using the soxhletation more effective in inhibiting the growth ofStaphylococcus aureus bacteria than the maceration method.

Metagenomic next-generation sequencing contributes to the diagnosis of mixed pulmonary infection: a case report

BackgroundPulmonary cryptococcosis (PC) and mixed pulmonary infection are difficult to be diagnosed due to the non-specificity and their overlapping clinical manifestations. In terms of the clinical diagnosis of PC and mixed pulmonary infection, conventional tests have limitations such as a long detection period, a limited range of pathogens, and low sensitivity. Metagenomics next-generation sequencing (mNGS) is a nascent and powerful method that can detect pathogens without culture, to diagnose known and unexplained infections in reduced time.Case presentationA 43-year-old female was admitted to the hospital after suffering from a cough for one month. At the time of admission, a contrast-enhanced chest CT revealed multiple nodules and plaques in her right lung, as well as the formation of cavities. The blood routine assays showed evidently increased white blood cell count (mainly neutrophils), CRP, and ESR, which suggested she was in the infection phase. The serum CrAg-LFA test showed a positive result. Initially, she was diagnosed with an unexplained pulmonary infection. Bronchoalveolar lavage fluid (BALF) samples were collected for microbial culture, immunological tests and the mNGS. Microbial culture and immunological tests were all negative, while mNGS detected Corynebacterium striatum, Pseudomonas aeruginosa, Streptococcus pneumoniae, and Cryptococcus neoformans. The diagnosis was revised to PC and bacterial pneumonia. Lung infection lesions were healed after she received targeted anti-infection therapy with mezlocillin and fluconazole. In a follow-up after 2 months, the patient’s symptoms vanished.ConclusionsHere, we demonstrated that mNGS was capable of accurately distinguishing Cryptococcus from M. tuberculosis in pulmonary infection, and notably mNGS was capable of swiftly and precisely detecting pathogens in mixed bacterial and fungal pulmonary infection. Furthermore, the results of mNGS also have the potential to adjust anti-infective therapies.

Rapid detection of four pathogens in bloodstream infection by antimicrobial peptide capture combined with multiplex PCR and capillary electrophoresis

Bloodstream infection (BSI) is a common and serious infection that threatens the life of patients. In this study, a rapid method combining magnetic separation techniques and multiplex PCR-based capillary electrophoresis (mPCR-CE) was developed to detect four pathogens causing BSI. Taking advantage of the binding capacity with the cell membrane, the antimicrobial peptides melittin was connected to magnetic beads as a broad-spectrum bacterial capture element, which was used to capture pathogenic bacteria in whole blood. The detection limit of this proposed method was significantly lower than that of the common PCR method, reaching 101 CFU/mL with excellent specificity. The overall clinical sensitivity was 90 %, and the clinical specificity was 100 %. There were no false positives for testing another 100 clinical samples suspected of bloodstream infections. The detection platform took only 3–4 h, which is obviously shorter than the identification time of the traditional methods. Our method possesses tremendous clinical application potential for its advantages in detecting mixed bacterial infections.