Bacterial Culture Research Articles

Bacillus subtilis grown in a "breathing" vessel without sparger aeration.

Here we present a "breathing" vessel consisting of expanded polytetrafluoroethylene, which allows gas exchange but no liquid permeation. The bacterial culture inside needs only agitation to promote air supply. Using this setup, a Bacillus subtilis cell factory for scyllo-inositol production grew to produce scyllo-inositol efficiently. The results indicate that our approach represents a sustainable "greener" approach for the cell factory.

Diagnostic Accuracy of Dermatoscopy Versus Microbiological Culture and Polymerase Chain Reaction in the Diagnosis of Onychomycosis: A Cross-Sectional Study.

Several clinical signs in dermatoscopy are very characteristic of onychomycosis and can be a quick complement for the diagnosis of onychomycosis. The aim of this study was to evaluate the diagnostic accuracy of dermatoscopy compared to microbiological culture and polymerase chain reaction (PCR), as well as the clinical signs associated with onychomycosis. The clinical signs of 125 patients were assessed cross-sectionally using dermatoscopy, and a positive or negative result was assigned. A sample was then taken for PCR and microbiological culture. Of the 125 patients, 69.6% (87/125) had positive results when both laboratory tests were combined. When they were not combined, the prevalence was lower at 48% (60/125) with PCR and at 43.2% (54/125) with culture. Furthermore, 76.8% (96/125) were classified as positive with dermatoscopy with a sensitivity of 1, a specificity of 0.76, positive predictive value of 0.91 and negative predictive value of 1 (with 95% confidence intervals). Of the 96 dermatoscopy-positive samples, 36 were negative with PCR (p < 0.001), 42 were negative with culture (p < 0.001) and nine were negative when both tests were combined (p < 0.001). Clinical signs that were significantly associated with the presence of onychomycosis were subungual hyperkeratosis (dermatoscopy: p = 0.004, odds ratio (OR) = 2.438; PCR + microbiological culture: p = 0.004, OR = 3.221), subungual detritus (p = 0.033, OR = 3.01, only with dermatoscopy) and dermatophytoma (dermatoscopy: p = 0.049, OR = 3.02; PCR + microbiological culture: p = 0.022, OR = 2.40). The results suggest that dermatoscopy is a good tool for the diagnosis of onychomycosis but should be used as a complementary test or for screening patients to be sampled for laboratory testing. The combination of the three tests can lead to a reduction of false-positive and false-negative clinical and laboratory results. This allows for early diagnosis and specific treatment based on test results.

Diarrhea and cholera surveillance for early warning and preparedness to prevent epidemics among Rohingya Myanmar nationals in Cox's Bazar, Bangladesh

Diarrheal diseases, especially cholera, can be a serious threat to Rohingya refugees in Cox's Bazar due to overcrowding and inadequate hygiene infrastructure. Assessing the risk, cholera surveillance network was established with the aim to identify the outbreak of diarrhea and cholera and help to take appropriate preventive measures including a vaccination campaign. The surveillance network has been ongoing for 6 years (2017–2023) in 17 health facilities. Diarrhea patients from Rohingya Myanmar nationals matched with case definition were enrolled and stool samples were tested by Rapid diagnostic test (RDT) for early cholera detection Multiple Logistic regression models were fitted to examine the associations of risk factors among cholera cases. A total of 17,252 stool samples were collected through surveillance. Among the tested samples, 588 (3.5 %) were detected positive by RDT, and 239 (1.4 %) Vibrio cholerae were isolated by microbiological culture. Between 2021 and 2023, the number of culture-confirmed cases exceeded that in the period from 2017 to 2020. In addition to V. cholerae; high positivity was identified for ETEC (11.8 %) followed by Salmonella (3.9 %) and Shigella (2.7 %). Most of the cholera cases were presented with vomiting, dehydration and loose watery and rice watery nature of stool (p value = <0.001). Major risk factors for cholera were 2–4 years age group (OR = 5.72; 95 % CI, 3.84–8.53.14; P = .001), process of water treatment (OR = 1.54; 95 % CI, 1.01–2.37; P = .046) and hand washing with soap before taking meals (OR = 0.6; 95 % CI, 0.39–0.92; P = .020. This study highlights the epidemiology of cholera among the Rohingya population and underscores the effectiveness of integrating surveillance data with early warning, alert, and response systems (EWARS) system, along with oral cholera vaccine (OCV) campaigns, in preventing major cholera outbreak.

The value of current diagnostic techniques in the diagnosis of fracture-related infections: Serum markers, histology, and cultures

Although fracture-related infection (FRI) is a serious complication following bone fractures, a comprehensive definition and diagnostic criteria have only emerged in recent years. According to this consensus definition, the diagnosis of FRI is based on preoperative and intraoperative suggestive or confirmatory criteria. Serum markers, histology, and microbiological cultures are considered to play a crucial role in the FRI diagnostic pathway. However, at the time of publication of the FRI consensus definition in 2018 and its update in 2020, limited data was available on the accuracy of these diagnostic methods. This review aims to provide an overview of recent publications and discuss whether new evidence has been obtained regarding the value of these current diagnostic techniques.Meanwhile, several studies have confirmed the limited prognostic value of C-reactive protein, erythrocyte sedimentation ratio, and white blood cell count. Other serologic markers for preoperative diagnosis of FRI with promising diagnostic performance are d-dimer, plasma fibrinogen, platelet count to mean platelet volume ratio, and a risk prediction model that includes soft tissue injury type and fracture complexity in addition to blood markers. However, their true diagnostic value in daily clinical practice needs to be investigated in further studies. Data on histology in FRI diagnosis is still limited, but its potential as a confirmatory criterion seems to lie in its high specificity. Recent studies indicate that tissue culture exhibits moderate sensitivity and high specificity, with sensitivity improvements achieved by sampling of five specimens and long-term culture. Implant sonication also appears to enhance the sensitivity of culture and the detection rate of polymicrobial infections.In conclusion, the true value of diagnostic techniques is difficult to assess, in part because it is measured against a gold standard that is itself imperfect and still evolving, but also because of methodological differences in sample processing or the use of different thresholds. Nevertheless, this review has identified that the value of current diagnostic techniques is high when used in combination. To draw more accurate conclusions about the value of serum markers, histology, and culture including sonication, future studies should be prospective and utilize a greater standardization in sampling and methodological protocols.

Pseudomonas mosselii: a potential alternative for managing pyrethroid-resistant Aedes aegypti.

Aedes aegypti is a widespread mosquito in tropical and subtropical regions that causes significant mortality and morbidity in humans by transmitting diseases, such as dengue fever and Zika virus disease. Synthetic insecticides, such as pyrethroids, have been used to control Ae. aegypti, but these insecticides can also affect nontarget organisms and contaminate soil and water. This study aimed to investigate the mosquitocidal activity of Pseudomonas mosselii isolated from pond sludge against larvae of Ae. aegypti. Based on the initial results, similar time-course profiles were obtained for the mosquitocidal activity of the bacterial culture and its supernatant, and the pellet resuspended in Luria-Bertani (LB) medium also showed delayed toxicity. These results imply that the toxic component can be released into the medium from live bacteria. Further research indicated that the toxic component appeared in the supernatant approximately 4 h after a 3-mL stock was cultured in 200 mL of LB medium. The stabilities of the P. mosselii culture and supernatant stored at different temperatures were also evaluated, and the best culture stability was obtained at 28 °C and supernatant stability at 4 °C. The bacterial culture and supernatant were toxic to larvae and pupae of not only susceptible Ae. aegypti but also pyrethroid-resistant strains. This study highlights the value of the mosquitocidal activity of P. mosselii, which has potential as an alternative insecticide to control pyrethroid-resistant Ae. aegypti in the field. © 2024 Society of Chemical Industry.

Multi-omics approach for understanding the response of Bacteroides fragilis to carbapenems

BackgroundThe prevalence of Bacteroides fragilis isolates resistant to first-line beta-lactam drugs is increasing, resulting in reduced treatment efficacy. Investigating the bacterial transcriptome and proteome can uncover links between bacterial genes and resistance mechanisms. In this study, we experimentally assessed in vitro the transcriptional and proteomic profiles of B. fragilis exposed to SICs of meropenem, an effective antimicrobial agent, collected from patients with intra-abdominal diseases at Astana City Hospital, Kazakhstan. MethodsB. fragilis was cultured in brain heart infusion broth and sub-cultured every 48 h for 8 days in media with and without meropenem. Total RNA was extracted from the liquid cultures using a commercial RNeasy mini kit, and strand-specific RNA sequencing (RNA-seq) was performed on the DNBSEQ platform. Raw RNA-seq data were retrieved from BioProject No. PRJNA531645 and uploaded to the NCBI Sequence Read Archive (accession no. SRX22081155). Proteins of B. fragilis were extracted and separated using sodium dodecyl sulphate–polyacrylamide gel electrophoresis, followed by analysis of the eluted peptides using liquid chromatography–tandem mass spectrometry. Cluster analysis utilised the Database for Annotation, Visualization, and Integrated Discovery. ResultsThe subinhibitory concentration (SIC) of meropenem was determined to be 0.5 μg/L (minimum inhibitory concentration: 1). Mapping of reads to the reference genome identified 2,477 expressed genes in all B. fragilis BFR KZ01 samples. Ten differentially expressed genes (DEGs) were common across comparison groups during and post-antibiotic exposure (wMEM vs. MEM2 and MEM2 vs. rMEM8); however, no substantially enriched Gene Ontology terms were identified. The cluster analysis highlighted a significant enrichment cluster (W-0560 oxidoreductase) of DEGs following antibiotic withdrawal. In total, 859 B. fragilis proteins were identified, with the expressions of three proteins, 3-oxoacyl-[acyl carrier protein] reductase, acetyl-CoA carboxylase biotin carboxylase subunit, and beta-ketoacyl-ACP synthase III, being upregulated in the enriched protein folding category. Notably, chaperone proteins such as FKBP-type peptidyl-prolyl cis-trans isomerases (involved in the cis-trans isomerisation of prolyl peptide bonds) and GroES (a co-chaperone functioning with GroEL) were also identified. ConclusionsUnder the influence of low doses of antibiotics defense mechanisms are activated which contribute to the emergence of resistance. These results provide insight into the response of B. fragilis to meropenem exposure, mainly at the SIC, contributing to the understanding bacterial survival strategies under stress conditions.

Survival of the probiotic strain Lacticaseibacillus paracasei subsp. paracasei F19 in high-hopped beers

This study aims to enhance understanding of probiotic lactic acid bacteria (LAB) survival in high-hopped beer formulations and their interactions with different yeasts and highlights the fermentation processes, microbial metabolism, and production of distinctive beer flavors. For this, this research used Lacticaseibacillus paracasei F19 (F19), Saccharomycodes ludwigii, and Saccharomyces cerevisiae strains US-05 (US-05) and Kveik (Kveik) for brewing. Bacterial and yeast cultures were prepared, fermented in wort, and analyzed in different hop concentrations (International Bitterness Units − IBU 0, 20, 40). Methods included physicochemical analysis, yeast and bacterial counts, RT-qPCR for gene expression, statistical analysis, and sensory evaluation by sommeliers following BJCP guidelines. Physicochemical analysis showed efficient fermentation across all hop concentrations (IBU 0, 20, 40), with decreasing SG and pH over time due to lactic acid bacteria and yeast metabolism. Higher hop levels (IBU 20 and 40) resulted in less acidic beer, indicating hop interference with bacterial activity. Yeast populations remained stable regardless of hop content, with Saccharomyces cerevisiae and Saccharomycodes ludwigii performing well. Probiotic strain F19 exhibited robust viability in all formulations. Sensory analysis favored higher hop content beers, suggesting consumer acceptance and potential health benefits of probiotic, high-hop beers. Higher hop content hindered sour beer production as only hop-free beers reached low pH levels. Probiotic strain F19 remained viable under high IBU formulations (20 and 40), with these being preferred by sommeliers using BJCP methodology. All yeast strains supported F19 survival. Further studies are needed on gastrointestinal resistance and clinical benefits.

Evaluation of mycobacterial microscopy and culture results from clinical samples: A five-year analysis.

Ehrlich-Ziehl-Neelsen (EZN) staining and culture methods are often used to diagnose tuberculosis. This study aimed to determine the acidfast bacteria (AFS) positivity rates in various clinical samples sent to our laboratory over five years and the growth and resistance rates in two different (solid and liquid) cultures and compare them with the data from Türkiye and the world. A total of 62.456 clinic samples were accepted in the microbiology laboratory between 2019 and 2024. The mycobacterial culture was performed by searching for acid-resistant bacilli microscopically and parallel inoculation media [solid Löwenstein-Jensen (L-J) and MGIT 960 liquid]. Those growing in the MGIT 960 system were identified using BD MGIT TBC Identification test kits that detect the MPT64 antigen. AFS and MPT64 antigenpositive samples were identified as Mycobacterium tuberculosis complex (MTBC) while AFS-positive samples and MPT64 antigen-negative results were classified as non-tuberculous mycobacteria (NTM). Drug susceptibility testing was performed with the BACTEC MGIT 960 SIRE kit. Susceptibility to NTM samples was not performed. Out of a total of 120.829 samples, 95.101 were lung samples and 25.728 were extrapulmonary samples. AFS positivity was detected in 2961 (2.4%) samples. MTBC grew in 6854 (5.6%) samples, and NTM grew in 1506 (1.24%) samples. Contamination was detected in 7171 (5.9%) media. Two thousand one hundred and sixty-nine susceptibility tests were performed. Considering antibiotic resistance rates, isoniazid resistance was detected in 154 (7%), rifampicin resistance in 140 (6.4%), ethambutol resistance in 18 (0.8%), and streptomycin resistance in 120 (0.5%) samples. All four-drug resistance was observed in 91 (4.1%) samples. AFP positivity and resistance rates for rifampicin have decreased significantly, while there have been no significant changes in NTM rates over the years. When our data was determined, the sensitivity of microscopy was low. It is understood that mycobacterial culture and microscopy must be evaluated together to exclude tuberculosis infection. The high mycobacterial culture positivity rate, which is 5.6%, is due to the high number of follow-up patients and new referrals. It is seen that the change in sensitivity rates is due to the period of the COVID-19 epidemic, and it is similar to World Health Organization (WHO) data.

Effects of the Anti-Inflammatory Properties of Bitter Leaf (Vernonia amygdalina) Ethanolic Extract on Streptocuccus Pyrogenes Induced Wister Rats

Aim: To investigate the effectiveness of V. amygdalina leaf extract on the remediation of the cytological and physiological damages on albino rats caused by streptococcus pyogenes. Study Design: Randomized Controlled Trial (RCT) design with multiple treatment groups. Research was done at the department of Science Laboratory Technology, University of Calabar, Cross River state, between March 2023 and June 2023. Methodology: Bitter extract was obtained through Soxlet extraction, and screened for the presence of secondary metabolites using standard phytochemical screening procedures. A pure culture of S. pyogenes was gotten from Microbiology Laboratory University of Calabar. Thirty five albino rats (100-175g), obtained from the Pharmacology department, University of Calabar were placed in 7 groups (A-G). Rats in groups A-F each was injected with the bacterium, observed for five days. Group A-E rats were subsequently treated with 0g/200 ml, 20g/200 ml, 40g/200 ml, 60g/200 ml, and 100g/200ml crude extract of bitter leaf respectively. While group F was treated with standard antibiotics. 0.5ml Cefuroxime Axetil tablets usp was injected into the rats. Group G rats served as negative control. Results: Results showed the presence of alkaloids, acidic compounds, tannins, flavonoids and Saponins. Haematocrit increased slightly in group A-E (18.3-46.3%) treated with 0g/200ml, 20g/200ml, 40g/200 ml, 60g/200 ml, and 100g/200ml concentrations of the extract, respectively. But more in group F (50.2%) that received antibiotics treatment, when compared with the control. Similar trend was observed for haemoglobin (Hb) values (15.3g/dl) and red blood cell (RBC) counts (3.0 x1012/l) that increased with increasing concentration of 100g/200ml extract treatment and antibiotics treatment (Hb = 15.7g/dl; RBC = 3.3 x1012/l) respectively. The white blood cell counts decreased with antibiotic treatment. Conclusion: Study established that in vivo administration of bitter leaf extract on albino rats was best at 60g/200 ml, and 100g/200ml in ameliorating haematotoxicity caused by S. pyogenes.

Comprehensive model for predicting toxic equivalents (TEQ) reduction due to dechlorination of polychlorinated dibenzo-p-dioxin and dibenzofurans (PCDD/F congeners)

Remediation-focused predictive tools for polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) rely on transformation models to evaluate the reduction in total contaminant load and toxic equivalency (TEQ). In this study, a comprehensive model predicting the profiles of PCDD/F congeners and the associated TEQs was developed. The model employs first-order kinetics to describe the transformation of 256 reactions for 75 PCDD congeners and 421 reactions for 135 PCDF congeners. It integrates the growth of anaerobic microbial guilds using Monod kinetics on hydrogen release compounds and stoichiometric growth for Dehalococcoides sp. The effects of temperature, salinity, pH, and availability of vitamin B12 (a cofactor) were also integrated. The PCDD/F congeners model was used to extract the first-order dechlorination rate constants from a number of pure culture and mixed microbial microcosm studies. Simulations for the transformation of PCDD/F congeners at concentrations representative of the Tittabawassee or Saginaw Rivers and watershed in MI, USA were carried out. For a starting TEQ of 5000 ng per kg dry sediment (ppt), the model predicted a decrease in the overall TEQ to below 2000 ppt after 2.6 years and below 250 ppt after ∼21 years. The developed model may be used for extracting rates from microcosm studies and to evaluate the effect of engineering interventions on TEQ reduction.

Enhanced γ-aminobutyric acid production by Co-culture fermentation with Enterococcus faecium AB157 and Saccharomyces cerevisiae SC125

γ-aminobutyric acid (GABA), a non-protein amino acid, is a major inhibitory neurotransmitter within the human nervous system. Research on enhancing GABA production through the co-culture of yeast and lactic acid bacteria is limited, and the intricacies of their interactions and the mechanisms behind GABA enhancement are not fully elucidated. In this study, five yeast strains from Sichuan Pao Cai were co-cultured with Enterococcus faecium AB157 to increase GABA content, and it was found that combining E. faecium AB157 with Saccharomyces cerevisiae SC125 at a 4:1 ratio, along with 12 g/L MSG, at 35 °C, optimized GABA production to reach levels of 6.57 g/L. Within the co-culture system, there was a notable increase in both glutamate decarboxylase activity (GAD) and medium pH, while the activity of Autoinducer-2 (AI-2) remained stable. Furthermore, the cell-free supernatants (CFS) from S. cerevisiae SC125 were able to enhance GABA production by E. faecium AB157, up to 4.61 g/L. RNA sequencing revealed that the GABA biosynthetic pathways and quorum sensing (QS) in co-fermentation on E. faecium AB157 were upregulated, and the ATP synthesis and arginine deiminase pathway were downregulated compare to the pure culture of E. faecium AB157.

Assessing Diagnostic Performance of Molecular Culture for Neonatal Sepsis: Protocol of the CHAMPIONS Study.

Managing neonatal sepsis is challenging due to nonspecific clinical signs, hematological markers with poor accuracy, and a lengthy turnaround time for the identification of microorganisms. Delaying the initiation of antibiotics in truly infected infants can lead to severe morbidity and mortality. Therefore, decisions regarding empiric antibiotic treatment are risk stratified, which exposes many uninfected infants to antibiotics. This causes gut microbiota perturbation, unnecessary hospital admissions, and the generation of multi-resistant organisms. High-speed diagnostic assays could expedite discontinuation or avert the initiation of antibiotics in uninfected infants. This study will evaluate the diagnostic performance of molecular culture (MC), a rapid broad-range PCR-based bacterial profiling technique, for diagnosing neonatal sepsis in infants below 90 days old. A multi-center prospective observational cohort study will include infants evaluated for early and late-onset sepsis. Routine evaluation for suspected sepsis includes microbiological cultures of blood. Additionally, blood for MC will be collected. For early-onset sepsis, umbilical cord blood may be used alternatively. Primary outcome is the agreement between MC and conventional blood culture results. Secondary outcome is the agreement of both assays with clinical sepsis using four different, commonly used definitions. Faster diagnostic pathways for sepsis may reduce antibiotic exposure time. Broad-range molecular assays may identify pathogens undetectable by conventional methods. Employment of umbilical cord blood samples for early-onset sepsis diagnosis can resolve challenges in collecting adequate blood volume and could further expedite treatment decisions.

Chromoblastomycosis: A Single Centre Clinical Laboratory Experience of Seven Years (2016-2022) and Literature Review From Pakistan.

Chromoblastomycosis (CBM) is a chronic infection of skin and subcutaneous tissue. CBM cases have been reported in local literature from Pakistan with heterogenous demographic, diagnostic and therapeutic information. The objective of this study is to share the experience of CBM from a large tertiary care hospital laboratory in Pakistan. This was a retrospective observational study. Histopathology and microbiology data of suspected CBM between 2016 and 2022 was retrieved. Patients' demographics, site of involvement, histopathological findings and positive microbiology cultures were assessed. Literature search on Google Scholar, PubMed and PakMediNet was done between 1990 and 2023 with multiple terms. A total of 16 CBM cases were identified; 14 were histopathology positive and two were both histopathology and culture positive. The median age was 21 years, and 11 patients were male. The predominant site was lower extremities followed by the face. Severe acanthosis, hyperkeratosis and granuloma with sclerotic bodies were observed in all histopathology slides. Alternaria spp. and Phialophora spp. were isolated from two culture-positive cases. A total of nine cases of CBM were reported from Pakistan in PubMed non-indexed journal. CBM is not a commonly thought of disease when evaluating skin lesions in Pakistan. A high index of suspicion when assessing patients who may have a history of trauma, exposure to soil and suggestive lesions is reasonable. An integrated approach between clinicians, histopathologist and microbiologist is required to do early identification and therapeutic interventions.

204. ALTERATIONS IN LOCAL OESOPHAGEAL MICROBIOTA AND OUTCOMES AFTER OESOPHAGECTOMY FOR OESOPHAGO-GASTRIC CANCER: A PILOT STUDY

Abstract Oesophageal cancer is a global health burden, with an estimated 600,000 new cases and 550,000 deaths per year. It is the 9th most common cancer yet is ranked fifth for cancer mortality. The current gold standard curative strategy includes oesophagectomy, which is a morbid procedure. In particular, anastomotic leaks are a feared complication postoperatively that result in significant morbidity. Studies of patients undergoing colorectal cancer surgery show that alterations in the local microbiota can contribute to complications including anastomotic leak. Furthermore, some pathogenic bacterial species have been implicated in shorter recurrence-free survival periods after curative oncologic surgery. Unlike bacterial culture, which requires viable bacteria to be preserved from the time of sampling through to processing, 16S RNA sequencing can be performed on both fresh and fixed tissues. However, there are few studies using either tissue source and to date, none that compare results from the same patient. The aims of this pilot study were to: 1. Assess the feasibility of testing archived histological specimens (i.e. formalin fixed) that in turn facilitates retrospective analysis of patients with known clinical outcomes (i.e. anastomotic leaks) 2. Identify any difference (variance, reduce or over-abundance) between the microbiota of patients who suffered anastomotic leak and those who did not 3. Generate preliminary data to inform sample sizes, statistical methods, workflow and cost of a future prospective trial Using an established Upper GI Cancer Surgery Database, we performed propensity score matching to identify 15 patients who had a clinically evident anastomotic leak to be matched with 15 patients who did not have an anastomotic leak. Known predictors of anastomotic leak (e.g. neoadjuvant radiotherapy, smoking status, COPD, chronic kidney disease, diabetes mellitus, chronic liver disease) have been controlled for. Fresh frozen and formalin-fixed samples for the same patients were available. The formalin-fixed tissue was comprised of the "anastomotic doughnuts" created at the time of the circular stapled anastomosis. This tissue was chosen because of its immediate proximity to the anastomosis. All samples were analysed using 16S RNA sequencing to obtain a full microbial profile. Sequencing data was analysed by scientific investigators blinded to the clinical outcome (i.e. anastomotic leak vs. no leak) using a pre-existing workflow and bioinformatics pipeline.

The emergence of parvovirus B19 as a pathogen in acute encephalitis syndrome.

Despite scarcity of data, in recent years, human parvovirus B19 (PVB19) has been emerging as an important pathogen in acute encephalitis syndrome (AES). But, PVB19 virus is mostly looked for only after the exclusion of other common pathogens implicated in AES. Hence, this study was conducted to correlate clinical, radiological, and sequencing data to establish the crucial role of PVB19 in AES. Cerebrospinal fluid and/or serum samples were collected from AES patients as per WHO criteria and tested by ELISA, real-time PCR and bacterial culture sensitivity for various pathogens. PVB19 positive samples were subjected to sequencing. PVB19 attributed to 5% of total AES cases in the present study with fatalities in two of eight cases. Two isolates of PVB19 belonged to Genotype 1 A whereas one belonged to Genotype 3B. On multivariate analysis of predictive symptoms of PVB19 AES cases, blurring of vision (odds ratio [OR] 20.67; p = 0.001) was found to be significant independent predictor of PVB19 AES. Six of eight patients (two encephalitis specific and four nonspecific) had abnormal radiological findings. Hence, being an emerging viral pathogen, PVB19 should be included in the diagnostic algorithm of AES for prompt diagnosis and definitive management to prevent undesired neurological sequelae.

CYSTOCENTESIS AND URINALYSIS IN ZOOMEDICINE: AN UNDERESTIMATED TOOL FOR LARGE FELID STANDARD HEALTH CHECKS.

Chronic kidney disease (CKD) is a prevalent disease among felids; yet its origin is still poorly understood, and the disease often remains asymptomatic for years, underscoring the need for early diagnosis. This study aimed to investigate the diagnostic value of urinalysis in accurately staging CKD, particularly as routine health checks in large felids often overlook its significance. In this research, ultrasound-guided cystocentesis (UGC) was performed on 50 captive nondomestic felids during routine veterinary health checks under general anesthesia. Urinalysis included microscopic examination of the sediment, measurement of urine specific gravity (USG) and protein to creatinine ratio (UPC). Additional serum kidney markers, such as creatinine and symmetric dimethylarginine, were compared with USG and UPC to assess their diagnostic value as urinary biomarkers. The results demonstrated proteinuria (UPC > 0.4) or borderline proteinuria (UPC 0.2-0.4) in 49% of the animals. Among these cases, 62% were of renal origin, and 38% were postrenal causes. USG was significantly higher in felids with borderline proteinuria compared to those with proteinuria. A moderate, but significant negative correlation between serum parameters and USG was observed, emphasizing the importance of assessing both diagnostic parameters during kidney evaluations. Additionally, felids with CKD have an increased risk of urinary tract infections, necessitating microscopic urinalysis and bacterial culture analysis. Abnormalities, including hematuria, pyuria, crystalluria, and bacteriuria, were found in approximately 38% of cases through microscopical examination of urine. No complications associated with UGC were observed and abnormal findings were detected in 60% of the cases. Based on these results, the authors recommend the inclusion of UGC and urinalysis as standard diagnostic tools in general health checks for nondomestic felids. This approach provides valuable insights into the early detection and staging of CKD, supporting early intervention and supportive medical care to prolong renal health in these animals.

Set Up and Utilization of a Three-Dimensional In Vitro Bioreactor System for Human Intestinal Studies and Microbial Co-Cultures.

The study of human intestinal physiology and host-microbe interactions is crucial for understanding gastrointestinal health and disease. Traditional two-dimensional cell culture models lack the complexity of the native intestinal environment, limiting their utility in studying intestinal biology. Here, we present a detailed protocol for the set up and utilization of a three-dimensional (3D) in vitro bioreactor system for human intestinal studies and bacterial co-culture. This article outlines the design and assembly of the bioreactor system, scaffold fabrication, bacterial culture techniques, analysis methods, and troubleshooting tips. By providing step-by-step instructions, the goal is to enable other laboratories to utilize physiologically relevant tissue models of the human intestine, incorporating key features, such as nutrient flow, multiple human cell types, 3D architecture, and microbial communities. The incorporation of commensal bacteria into the bioreactor system allows for the investigation of complex host-microbe interactions, providing insight into gastrointestinal health and pathology. This article serves as a comprehensive resource for scientists seeking to advance their understanding of intestinal biology toward the development of novel therapeutic strategies for gastrointestinal disorders. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Scaffold design Basic Protocol 2: Intestinal cell culture: Caco2 cells Basic Protocol 3: Intestinal cell culture: organoids Basic Protocol 4: Bioreactor design and set up Basic Protocol 5: Bacteria in 3D bioreactor set up Basic Protocol 6: Bacteria and drug dosing.

No sex-dependent mortality in an amphibian upon infection with the chytrid fungus, Batrachochytrium dendrobatidis.

One of the major factors driving the currently ongoing biodiversity crisis is the anthropogenic spread of infectious diseases. Diseases can have conspicuous consequences, such as mass mortality events, but may also exert covert but similarly severe effects, such as sex ratio distortion via sex-biased mortality. Chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd) is among the most important threats to amphibian biodiversity. Yet, whether Bd infection can skew sex ratios in amphibians is currently unknown, although such a hidden effect may cause the already dwindling amphibian populations to collapse. To investigate this possibility, we collected common toad (Bufo bufo) tadpoles from a natural habitat in Hungary and continuously treated them until metamorphosis with sterile Bd culture medium (control), or a liquid culture of a Hungarian or a Spanish Bd isolate. Bd prevalence was high in animals that died during the experiment but was almost zero in individuals that survived until the end of the experiment. Both Bd treatments significantly reduced survival after metamorphosis, but we did not observe sex-dependent mortality in either treatment. However, a small number of genotypically female individuals developed male phenotype (testes) in the Spanish Bd isolate treatment. Therefore, future research is needed to ascertain if larval Bd infection can affect sex ratio in common toads through female-to-male sex reversal.

Predictors of Sepsis and Sepsis-related Mortality in Critically Ill Burn Patients: A Single Tertiary Care Center Experience.

Clinical diagnosis of sepsis is challenging, emphasizing the importance of regular bacterial surveillance, and tailored antimicrobial therapy. This study aims to elucidate the predictors of sepsis in critically ill burn patients. A retrospective analysis was conducted on patients admitted to the burn intensive care unit between 2016 and 2022. Demographics, type of burn, total body surface area (TBSA), presence of inhalation injury, mortality, sepsis, deep vein thrombosis, pulmonary embolism, pneumonia, cultures, and laboratory findings were collected. Descriptive statistics and survival analysis were used to analyze trends during the 7-year period. The study encompassed 196 participants. Among patient factors, men constituted 73.4% (n = 102) of those without sepsis and 86.0% (n = 49) with sepsis, with an association between sepsis and lower age (34 versus 41 years) as well as larger TBSA (41.1% versus 17.3%). Inhalation injury was a significant predictor of sepsis [35.1% (n = 20) versus 11.6% (n = 16)]. Mortality was higher in sepsis cases [17.5% (n = 10) versus 2.9% (n = 4)], as well as positive blood cultures [47.4% (n = 27) versus 2.2% (n = 3)], positive wound cultures [71.9% (n = 41) versus 12.2% (n = 17)], and positive fungal cultures [12.3% (n = 7) versus 0% (n = 0)]. Multivariable analysis identified age and TBSA as significant predictors of sepsis (P = 0.025, P < 0.001). Age, TBSA affected emerge as a strong risk factor for sepsis among critically ill burn patients. It underscores the need for vigilant monitoring to improve outcomes and reduce sepsis-related mortality.

Unde venis? Bacterial resistance from environmental reservoirs to lettuce: tracking microbiome and resistome over a growth period.

Fresh produce is suggested to contribute highly to shaping the gut resistome. We investigated the impact of pig manure and irrigation water quality on microbiome and resistome of field-grown lettuce over an entire growth period. Lettuce was grown under four regimes, combining soil amendment with manure (with/without) with sprinkler irrigation using river water with an upstream wastewater input, disinfected by UV (with/without). Lettuce leaves, soil, and water samples were collected weekly and analyzed by bacterial cultivation, 16S rRNA gene amplicon sequencing, and shotgun metagenomics from total community DNA. Cultivation yielded only few clinically relevant antibiotic-resistant bacteria (ARB), but numbers of ARB on lettuce increased over time, while no treatment-dependent changes were observed. Microbiome analysis confirmed a temporal trend. Antibiotic resistance genes (ARGs) unique to lettuce and water included multidrug and β-lactam ARGs, whereas lettuce and soil uniquely shared mainly glycopeptide and tetracycline ARGs. Surface water carried clinically relevant ARB (e.g. ESBL-producing Escherichia coli or Serratia fonticola) without affecting the overall lettuce resistome significantly. Resistance markers including biocide and metal resistance were increased in lettuce grown with manure, especially young lettuce (increased soil contact). Overall, while all investigated environments had their share as sources of the lettuce resistome, manure was the main source especially on young plants. We therefore suggest minimizing soil-vegetable contact to minimize resistance markers on fresh produce.