Fluid Samples Research Articles

Real-time PCR versus culture of bronchoalveolar lavage fluid for detecting Bordetella bronchiseptica in dogs with persistent lower respiratory signs: a retrospective study of 23 cases.

Bordetella bronchiseptica (Bb) is a frequent pathogen in dogs and a component of the Canine Infectious Respiratory Disease Complex (CIRDC). However, it can also be detected in the lower respiratory tract of healthy animals, complicating its interpretation in clinical settings. This retrospective study compared the detection of Bb in bronchoalveolar lavage fluid (BALF) samples using real-time PCR versus bacterial culture in dogs referred for persistent lower respiratory tract signs to a referral hospital in Spain between 2018 and 2024. A total of 143 BALF samples were analyzed; 23 dogs tested positive for Bb by at least one method and were included in the clinical analysis. Real-time PCR identified Bb in 16.1% (23/143) of cases (95% CI:10.4-23.1%). Among these, 40% (9/23) had negative culture results. Co-infections with Mycoplasma spp. or Mycoplasma cynos were detected in 30% (7/23) of cases, and 26% (6/23) had positive cultures for other bacteria. Most dogs (91% (21/23)) had received antibiotics prior to BALF collection. The median time to clinical remission was 30 days. While real-time PCR showed a higher detection rate than culture, its results must be interpreted cautiously. PCR may amplify DNA from non-viable organisms or colonizers, and thus does not confirm active infection. Our findings underscore the need to integrate molecular, microbiological, and clinical data to reach an accurate diagnosis and avoid inappropriate antimicrobial use. Combining PCR and culture may enhance diagnostic accuracy, as culture alone would have failed to detect Bb in 40% of PCR positive dogs.

P28 Antibiotic susceptibility testing of coagulase-negative staphylococci species isolates from the Royal Orthopaedic Hospital, Birmingham

Abstract Background Coagulase-negative staphylococci (CNS) are often considered non-pathogenic; however, they can be a major cause of chronic prosthetic joint infections (PJIs), associated with significant morbidity, mortality, and healthcare costs in the UK. Oral antibiotic options are often limited due to resistance, making IV glycopeptide antibiotics (vancomycin and teicoplanin) the preferred first-line treatment. While vancomycin-resistant isolates are rare, susceptibility to teicoplanin can vary, with some studies indicating that resistance may occur in up to a quarter of cases. Teicoplanin once daily administration provides significant benefits for managing outpatient treatment of certain patients with PJIs, compared to vancomycin, which requires twice-daily dosing. Objectives The study aimed to assess local antibiotic susceptibility data in CNS over the past five years, particularly in patients with bone and joint infections from the Royal Orthopaedic Hospital, Birmingham. The secondary objective was to determine the teicoplanin, daptomycin, and linezolid susceptibility profile of the included isolates. Methods This retrospective study involved a search of the Laboratory Information Management System (Telepath) to identify all isolates of CNS, including Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis and Staphylococcus capitis from bone, tissue, or fluid samples obtained from patients who underwent operations at the Royal Orthopaedic Hospital in Birmingham, one of the largest centres of its kind in Europe. MALDI-TOF MS (bioMérieux) was used for rapid and accurate species identification, while VITEK2® (bioMérieux) was employed for antimicrobial susceptibility testing. Teicoplanin susceptibility was assessed using the Etest® teicoplanin (bioMérieux) and EUCAST breakpoints (≤4 = susceptible, >4 = resistant). Results We identified a total of 683 specimens that met the eligibility criteria. After excluding duplicate isolates and focusing on those that underwent a teicoplanin e-test, we included 47 isolates in the study. The median age of the patients was 69 years, including 29 males (62%) and 18 females (38%). The primary infection sites included the knee joint (36%) and hip joints (34%), followed by the shoulder, ankle joints, and miscellaneous specimens. S. epidermidis was the predominant species isolated from the analysed specimens (n=45, 96%). All isolates showed susceptibility to vancomycin, daptomycin, and linezolid. The highest antibiotic resistance recorded was for clindamycin (n=39, 83%), followed by trimethoprim (n=37, 79%) and fluoroquinolones (ciprofloxacin, 65%). Susceptibility testing for additional oral agents revealed significant resistance to tetracycline (n=28, 60%). and rifampicin (n=20, 42%). Results for teicoplanin susceptibility indicated that 16 (34%) isolates exhibited resistance based on EUCAST breakpoints. Among all isolates, 21 (45%) were sensitive to at least one oral option besides linezolid. Notably, we observed an increased rate of teicoplanin resistance in 2023 and 2024 compared to 2020 and 2022. The study did not assess the impact of antibiotic use or other factors that could contribute to antibiotic resistance. Conclusions S. epidermidis was the predominant species identified among CNS associated with bone and joint infections. Most isolates exhibited resistance to commonly prescribed oral antibiotics. Furthermore, there has been a growing resistance to teicoplanin over the years, necessitating the need for further studies to explore the underlying factors.

Amniotic and cervical fluids progranulin levels in pregnancies complicated by spontaneous preterm delivery with respect to intra-amniotic complications—a retrospective cohort study

The main aim of the study was to determine progranulin levels in amniotic and cervical fluid samples from pregnancies complicated by preterm prelabor rupture of membranes (PPROM) or preterm labor with intact membranes (PTL), with concomitant microbial invasion of the amniotic cavity and/or intra-amniotic inflammation. A total of 104 and 108 women with PPROM and PTL, respectively, were included. Paired amniotic and cervical fluid samples were obtained using transabdominal amniocentesis and Dacron polyester swabs, respectively. Progranulin levels were assessed with an enzyme-linked immunosorbent assay. Women with PPROM and PTL were divided into subgroups based on microbial invasion of the amniotic cavity and/or intra-amniotic inflammation. Differences in progranulin levels among the PPROM and PTL subgroups were found in amniotic fluid: (a) PPROM: intra-amniotic infection: 51.8 pg/mL, sterile intra-amniotic inflammation: 52.8 pg/mL, colonization: 36.4 pg/mL, and negative amniotic fluid: 35.0 pg/mL; p < 0.0001; (b) PTL: intra-amniotic infection: 75.3 pg/mL, sterile intra-amniotic inflammation: 54.0 pg/mL, and negative amniotic fluid: 39.1 pg/mL; p < 0.0001. The corresponding differences were not found in cervical fluid: (a) PPROM: p = 0.14; (b) PTL: p = 0.53. In conclusion, amniotic fluid progranulin levels increased in PPROM and PTL cases with concomitant intra-amniotic inflammation, regardless of whether microbial invasion of the amniotic cavity was present or absent.

Anti-NMDAR encephalitis triggered by EBV and HSV-1: A case report and literature review.

Anti-N-methyl-d-aspartate receptor (NMDAR) encephalitis is a central nervous system disorder driven by autoantibodies, characterized by a wide range of complex clinical manifestations. Although tumors, such as teratomas, and viral infections have been identified as potential triggering factors, the precise etiology of this condition remains to be elucidated. While herpes simplex virus type 1 (HSV-1) has been linked to the onset of anti-NMDAR encephalitis, cases involving coinfection with Epstein-Barr virus (EBV) and HSV-1 as causative factors are exceedingly rare. A 46-year-old female patient who exhibited mental abnormalities, epileptic seizures, and altered consciousness. Analysis of cerebrospinal fluid samples revealed the presence of EBV and HSV-1 infections. The initial autoimmune encephalitis antibody test returned negative results. However, a subsequent test indicated the presence of anti-NMDAR antibodies in the cerebrospinal fluid, whereas serum analysis remained negative. The patient was diagnosed with anti-NMDAR encephalitis resulting from infection with the EBV and HSV-1. Antiviral treatments were administered to the patient in conjunction with intravenous methylprednisolone and human immunoglobulin pulse therapy. This initial treatment did not yield satisfactory results, and the patient's condition remained critical, necessitating the introduction of rituximab for immunosuppressive therapy. Subsequently, the treatment regimen was adjusted to include intravenous tocilizumab, along with an intrathecal injection of 10 mg methotrexate and 10 mg dexamethasone. Ultimately, the patient's clinical symptoms improved relative to their previous condition, and repeated examinations demonstrated a gradual decrease in the white blood cell count in the cerebrospinal fluid, indicating that the treatment was effective. This case is the first diagnosed at our hospital and represents a rare instance in the literature of anti-NMDAR encephalitis directly triggered by co-infection with EBV and HSV-1. The diagnostic process emphasizes the need to maintain a high level of suspicion and repeat antibody tests in patients suspected of autoimmune encephalitis, even when initial antibody tests are negative.

Multimodal evaluation of cerebrospinal fluid from breast cancer leptomeningeal metastases using circulating tumor cell detection, single-cell sequencing, and proteomics

BackgroundPatients with human epidermal growth factor receptor 2 positive (HER2+) or triple-negative breast cancer are at higher risk than those with other breast cancer subtypes for developing brain metastases, including leptomeningeal metastases. Despite advances in the treatment of primary breast tumors, the prognosis of breast cancer patients with brain metastases remains poor. Frequent and sensitive monitoring of response to treatments is crucial to optimize treatments for brain metastases. Multimodal molecular analyses of cells in cerebrospinal fluid (CSF) may provide a less invasive approach than biopsy for monitoring disease in the central nervous system.MethodsA 41-year-old female with refractory leptomeningeal metastases from HER2 + breast cancer had serial sampling of cerebrospinal fluid (CSF) from an Ommaya reservoir prior to, and after, the addition of sacituzumab govitecan to her treatment regimen. Cytopathologic clinical analysis, circulating tumor cell enumeration, single-cell RNA sequencing, and cytokine profiling assays were performed on collected samples.ResultsA reduction in CSF tumor cells and absence of HER2-expressing cells was observed following treatment, with MRI confirming decreased tumor size. Single-cell RNA sequencing revealed diverse tumor and immune cell populations in CSF, highlighting gene expression signatures associated with breast cancer and triple negative disease and dynamic changes in cellular composition throughout the course of treatment. Cytokine profiling identified increased pro-inflammatory cytokines post-treatment.ConclusionThese findings highlight the potential utility of molecular profiling of CSF as an early indicator of treatment response and an approach for identifying resistance mechanisms and optimal combination therapies.

Increased cerebrospinal fluid and plasma apoE glycosylation is associated with reduced levels of Alzheimer’s disease biomarkers

The apolipoprotein E (APOE) ε4 allele is the strongest genetic risk factor for late-onset Alzheimer’s disease (AD). ApoE is glycosylated with an O-linked Core-1 sialylated glycan at several sites; however, the impact and function of this glycosylation on AD biomarkers remain unclear. We examined apoE glycosylation (total and secondary) in a cohort of cerebrospinal fluid (CSF, n = 181) and plasma (n = 178) samples from the Alzheimer’s Disease Neuroimaging Initiative (ADNI) stratified into 4 groups: cognitively normal (CN), Mild Cognitive Impairment (MCI), progressors, and non-progressors based on delayed word recall performance over 4 years. We observed decreasing glycosylation (reduced % of apoE being glycosylated) from apoE2 > apoE3 > apoE4 in the CSF and in plasma (apoE3 > apoE4), with stronger effect sizes for secondary glycosylation in CSF (total glycosylation in CSF: E2 > E3 (4.6%), E3 > E4 (5.1%), E2 > E4 (9.4%); secondary glycosylation in CSF: E2 > E4 (33.1%), E3 > E4 (25.4%); total glycosylation in plasma: E3 > E4 (24.2%). Secondary ApoE glycosylation was reduced (8%, p = 0.009) in the MCI group compared with the CN group and in the progressor group compared with the non-progressor group (7%, p = 0.01). In CSF, higher apoE glycosylation was cross-sectionally associated with lower total tau (t-tau) and p-tau181. In CSF, greater apoE4 glycosylation was associated with lower t-tau and p-tau181 levels. These results indicate strong associations between apoE glycosylation and biomarkers of AD pathology independent of apoE genotype, warranting a deeper understanding of the functional role of apoE glycosylation in AD tau pathology.

Protein-based Diagnosis and Analysis of Co-pathologies Across Neurodegenerative Diseases: Large-Scale AI-Boosted CSF and Plasma Classification.

Neurodegenerative diseases (including Alzheimer's disease, Parkinson's disease, Frontotemporal dementia, and Dementia with Lewy bodies) pose diagnostic challenges due to overlapping pathology and clinical heterogeneity. We leveraged proteomic data from more than 21,000 cerebrospinal fluid and plasma samples to develop and validate explainable, boosting-based multi-disease AI classifiers. The models achieved weighted AUCs in the testing datasets of 0.97 for CSF and 0.88 for plasma, equivalent to traditional biomarkers. The model was validated with neuropathological and clinical data, confirming robust generalizability without any retraining. Using zero-shot learning, we classified disease subtypes including autosomal dominant AD and prodromal PD and clarified disease states for those with conflicting clinical information. The model also showed the ability to prioritize cognitively normal individuals at disease risk. This framework enabled the identification and quantification of continuous, individual-level disease probabilities that allow for the quantification of overlap across diseases and co-pathologies within an individual. Through this work, we establish a benchmark computational framework for enhancing diagnostic precision in NDs.

Establishment of Human Lung Cancer Organoids Using Small Biopsy and Surgical Tissues

Background/Objectives: Lung cancer is a highly diverse disease, and reliable preclinical models that accurately reflect tumor characteristics are essential for studying lung cancer biology and testing new therapies. This study aimed to establish patient-derived tumor organoids (PDTOs) using small biopsy samples and surgical specimens to create a model system that preserves the genetic and histological features of the original tumors. Methods: PDTOs were generated from 163 lung cancer specimens, including 109 samples obtained using endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) or bronchoscopy, 52 surgical specimens, and 2 pleural fluid samples. The organoid establishment rate beyond passage three was assessed, and histological subtypes and genetic profiles were analyzed using immunohistochemical staining and targeted exome sequencing. Results: The overall PDTO establishment rate was 34.4% (56/163), and 44.6% (25/56) of these organoids retained the histological and genetic features of the parental tumors. Genetic analysis identified key mutations, including KRAS G12C, EGFR L858R, MET exon 14 skipping mutation, and ROS1 fusion. PDTOs successfully formed tumors in mice while maintaining the genetic characteristics of the original tumors. Co-culture of PDTOs with cancer-associated fibroblasts (CAFs) resulted in increased resistance to paclitaxel. In the co-culture model of PDTOs with immune cells, dose-dependent growth inhibition of PDTOs was observed in response to immune checkpoint inhibitors. Conclusions: PDTOs established from small biopsy and surgical specimens serve as a valuable model for studying lung cancer biology, tumor microenvironment interactions, and drug response. This model has the potential to improve personalized treatment strategies.

Utility of Multiplex PCR for Rapid Diagnosis of Acute Bacterial Meningitis: A North Indian Tertiary Care Experience.

Rapid diagnosis of acute bacterial meningitis (ABM) remains challenging. This study evaluated multiplex PCR versus conventional methods. A total of 1,248 cerebrospinal fluid samples from suspected ABM patients were tested using Gram stain, culture, and multiplex PCR targeting Streptococcus pneumoniae, Haemophilus influenzae type b, Neisseria meningitidis, Streptococcus agalactiae, and 16S rRNA gene. Multiplex PCR detected pathogens in 9.3% of samples, compared to 2.3% by culture and 1.5% by Gram stain. PCR identified H. influenzae (13), S. pneumoniae (7), N. meningitidis (1), S. agalactiae (1). With 90% sensitivity and 92.6% specificity, multiplex PCR is a valuable complementary diagnostic tool for ABM.

Cytomorphology of Renal Hydatid Cyst Mimicking as Simple Renal Cyst.

Echinococcosis is one of the zoonotic illnesses. Echinococcus granulosus larvae are the causative agents. Renal hydatid cysts are extremely rare and are typically unilateral. If they evolve slowly, they may not show any symptoms for years. Only 2%-3% of cases of hydatidosis have isolated renal hydatid cysts, making them rare. In this case, the radiological and clinical results suggested only a simple renal cyst, and cytopathology clinched the final diagnosis. A 20 mL turbid cyst fluid sample was sent for cytological analysis. Many scolices with radially arranged hooklets, scattered hooklets, macrophages, foreign body giant cells, and necro-inflammatory cell infiltrate were visible under the microscope. INSIDE THIS MONTH'S CYTOPATHOLOGY: In this study, the author highlights the cytology features of renal hydatidosis clinically and radiologically, mimicking a simple renal cyst. This case demonstrates the relevance of cytopathological screening in the early detection of unusual illnesses at rare locations. TWEETING (~280 CHARACTERS) THROUGH CYTOPATHOLOGY TWITTER HANDLE: A study of 24 #renal lesions found #FNA #fluid cytology on laparoscopic deroofing of the left renal cyst. Smears show singly scattered hooklets, intact scolices with radially arranged hooklets in a background of acellular debris and foreign body giant cells. #CytoJ #CytoPath #hydatid cyst #renal cyst #cytology.

A comparative study using Xpert MTB/RIF and culture methods evaluates MassARRAY technology for rapid detection of Mycobacterium tuberculosis and drug resistance

Tuberculosis (TB) remains a major global health threat, with the urgent need for rapid and accurate diagnostic methods to improve control and treatment outcomes. This study evaluates the performance of MassARRAY technology for detecting Mycobacterium tuberculosis (MTB) and identifying drug resistance, compared to traditional culture methods and Xpert MTB/RIF. From July 2021 to February 2024, bronchoalveolar lavage fluid (BALF) samples from 289 suspected pulmonary tuberculosis patients at Henan Provincial Chest Hospital, China, were tested using MassARRAY, Xpert MTB/RIF, and conventional culturing techniques. The performance of each method was assessed for MTB detection, and the ability of MassARRAY to identify drug resistance was compared with standard drug susceptibility testing (DST). MassARRAY demonstrated a sensitivity of 96.5% and a specificity of 34.6% for MTB detection, outperforming the Xpert MTB/RIF assay in sensitivity (94.7%) but showing lower specificity. In detecting rifampicin resistance, MassARRAY achieved concordance rates of 83.93% with Xpert MTB/RIF and 72.73% with DST. Furthermore, MassARRAY successfully identified key genetic mutations associated with drug resistance, such as rpoB 531 for rifampicin and katG 315 for isoniazid. MassARRAY demonstrated high concordance with DST for several drugs, including isoniazid, kanamycin, and streptomycin, but exhibited limitations in detecting resistance to pyrazinamide, clofazimine, cycloserine, and linezolid. Overall, MassARRAY provides a rapid, cost-effective, and high-throughput diagnostic platform for MTB and drug resistance, particularly for first-line anti-tuberculosis drugs. While limitations in specificity and resistance detection for certain second-line drugs exist, its ability to rapidly provide comprehensive resistance profiles makes it a valuable tool for TB management.

Performance Evaluation of the Sysmex XN, XR, and DI‐60 Body Fluid Applications for Leukocyte Differentiation

ABSTRACTIntroductionIn response to the growing demand for automation in body fluid (BF) analysis, we evaluated the research four‐part white blood cell (WBC) differentiation provided by the Sysmex XN and XR hematology analyzers (BF mode), as well as the Sysmex digital cell imaging analyzer DI‐60 (CellaVision) BF application for microscopic analysis.MethodsCerebrospinal, pleural, peritoneal, and synovial fluid samples were analyzed. The four‐part WBC differentiation (neutrophil, lymphocyte, monocyte, and eosinophil count (%)) provided by the XN‐ and XR‐BF mode and the DI‐60 BF application was compared to manual microscopic examination of May‐Grünwald Giemsa stained cytocentrifuge slides. Additionally, the DI‐60 BF application was assessed for its capability to detect malignancies.ResultsA total of 205 BF samples were included in the evaluation of the four‐part WBC differentiation of the XN‐ and XR‐BF mode. A very strong correlation coefficient (rs) between the XN/XR analyzer and manual microscopic examination was observed for neutrophils (%): 0.851 (XN); 0.862 (XR) and lymphocytes (%): 0.833 (XN); 0.842 (XR). The XN and XR analyzers reported lower monocyte counts compared to manual microscopic examination (bias = −3.21% (XN); −1.16% (XR)). The eosinophil count (%) showed suboptimal correlation between the automated analyzers and manual microscopy (rs = 0.428 (XN); 0.555 (XR)), probably due to low levels and outliers. Out of 161 cytospin slides, 39 could not be processed by the DI‐60, resulting in 122 remaining samples. All WBC types showed strong correlation between the manual count and the DI‐60 count (rs values of 0.735–0.974).ConclusionsThe four‐part WBC differentiation provided by the Sysmex XN‐ and XR‐BF mode and the Sysmex DI‐60 BF application are valuable tools for automating and standardizing BF analysis.

Benefits and challenges of host depletion methods in profiling the upper and lower respiratory microbiome

Metagenomic sequencing for respiratory pathogen detection faces two challenges: efficient host DNA depletion and the representativeness of upper respiratory samples for lower tract infections. In this study, we benchmarked seven host depletion methods, including a new method (F_ase), using bronchoalveolar lavage fluid (BALF), oropharyngeal swab (OP), and mock samples. All methods significantly increased microbial reads, species richness, genes richness, and genome coverage while reduced bacterial biomass, introduced contamination, and altered microbial abundance. Some commensals and pathogens, including Prevotella spp. and Mycoplasma pneumoniae, were significantly diminished. F_ase demonstrated the most balanced performance. High-resolution microbiomes profiling revealed distinct microbial niche preferences and microbiome disparities between the upper and lower respiratory tract. In pneumonia patients, 16.7% of high-abundance species (>1%) in BALF were underrepresented (<0.1%) in OP, highlighting OP’s limitations as lower respiratory proxies. This study underscores both the potential and challenges of metagenomic sequencing in advancing microbial ecology and clinical research.

Forensic analysis of mixed semen and vaginal fluids via ATR-FTIR spectroscopy for identification purposes in sexual aggression cases

BackgroundForensic science is an interdisciplinary field that applies principles and techniques from the natural and physical sciences to matters of law and criminal justice. As an academic discipline, forensic science encompasses a broad range of scientific methods used to analyze physical evidence from crime scenes to aid in investigations, legal proceedings, and the enforcement of justice. Within this field, forensic genetics plays a pivotal role by employing genetic material obtained from biological samples at crime scenes to establish associations between suspects and criminal acts, particularly in cases involving sexual assault. Furthermore, the accurate identification of biological fluids is integral to the advancement of forensic investigations. In the context of sexual offenses, spectroscopic techniques have gained prominence for their capacity to analyze body fluids (such as semen and vaginal fluid) on substrates taken from the victims’ body or at the scene rapidly and non-destructively, without sample destruction or the need for extensive sample preparation. Among these techniques, Fourier Transform Infrared (FTIR) spectroscopy stands out due to its ability to provide rapid, cost-effective, and non-invasive analysis while preserving the integrity and quantity of the biological specimen.ResultsThe present study investigates the application of FTIR spectroscopy for the identification of seminal fluid, vaginal fluid, and their combinations on two fabric substrates: white cotton and patterned nylon. Spectral analysis enabled the identification of distinct functional groups associated with each type of biological fluid. Spectral markers specific to vaginal fluid and mixed samples were successfully distinguished.ConclusionA notable finding of the study was the identification of the PSA/Fructose functional group as a biochemical marker indicative of seminal fluid within mixed samples containing vaginal secretions. Although both seminal and vaginal fluids were found to share five common biomolecular components, the presence of a distinct PSA/Fructose peak was characteristic of seminal fluid, whereas the detection of a glycogen peak—attributed to epithelial shedding from the cervix—was indicative of vaginal fluid.This distinction facilitated the clear differentiation between seminal and vaginal constituents within mixed biological samples. These results highlight the efficacy and reliability of Fourier Transform Infrared (FTIR) spectroscopy as a forensic tool for the identification of biological fluids, particularly in the context of sexual assault investigations.

Feeding amylolytic and fibrolytic exogenous enzymes in feedlot diets: effects on ruminal parameters, nitrogen balance and microbial diversity of Nellore cattle

BackgroundThe environmental impact of feedlot operations is a growing concern, as cattle excrete a significant portion of feed nutrients as waste. Exogenous feed enzymes (EFE) have gained interest for their potential to enhance feed efficiency in ruminants by improving nutrient digestion. However, EFE effects on ruminal parameters have shown inconsistencies, with limited research on nitrogen metabolism and rumen microbiome impacts. Moreover, the synergistic effects of combining different EFEs remain unclear. This study aimed to evaluate the effects of individual and combined EFE products in feedlot diets on ruminal fermentation parameters, nitrogen metabolism, and ruminal microbial communities. Ten rumen-cannulated Nellore steers [543 ± 28.6 kg of body weight (BW)] were distributed in a replicated Latin-square design (5 × 5) in individual pens. Treatments included: control (CON, no EFE supplementation), amylase [AML, 0.5 g/kg of diet dry matter (DM)], xylanase (FBL, 0.9 g/kg DM), half dose combination (HD, 0.25 g of AML + 0.45 g of FBL/kg of DM), and full dose combination (FD, 0.5 g of AML + 0.90 g of FBL/kg of DM). The experimental period lasted 19 d and included total urine and feces collection (d 15 to 18) and rumen fluid sampling (d 19) at 0, 4, 8, 12, and 16 h post-feeding for ammonia, volatile fatty acids (VFA), pH and microbiome analysis.ResultsEFE supplemented animals exhibited lower ruminal ammonia concentrations (P = 0.040), and higher acetate proportions (P < 0.001) compared to the control group. EFE supplementation resulted in reduced nitrogen (N) excretion in feces (P = 0.049) and urine (P = 0.036), contributing to improved N retention and efficiency (P = 0.045). Additionally, EFE products induced shifts in various microbial taxa at family and genera levels (P ≤ 0.10), which may be associated with the changes observed in ruminal fermentation.ConclusionsOur findings demonstrate that EFE supplementation enhances nitrogen retention, reduces ruminal ammonia, and alters ruminal fermentation profiles and microbial populations in feedlot cattle. While the expected synergism between amylase and xylanase did not significantly impact rumen fermentation parameters, it did induce shifts in the rumen microbiome. These results suggest that EFE supplementation may be a promising strategy for improving nutrient utilization and potentially reducing the environmental impact of feedlot operations.

Synthesis and Rheological Characterization of Magnetorheological Fluids

Magnetorheological (MR) fluids correspond to the kind of smart materials, capable of varying their rheological characteristics i.e., viscosity, yield stress, etc. on varying the magnetic field. In this article, 18 MR fluid samples utilizing various constituents are prepared and optimized with the aid of an L18 orthogonal array. The synthesized samples are experimentally tested on a rotational oscillatory shear rheometer namely MCR-102 (modular compact rheometer) to obtain the shear stress values at various shear rates (0 - 1000 Sec-1) and different currents (0 - 5 Ampere). This experimentally obtained data is fitted to the Herschel-Bulkley fluid model to estimate the On-state Yield Stress (OYS) for the tested MR fluid samples. An optimal set of the selected parameters i.e., silicon oil (carrier fluid), CI6 as the type of iron particle, 24 (Vol%) of iron particle proportion, and ethylene glycol monostearate (1.0% by volume) has been obtained using Taguchi analysis. An experimental verification test on the MR fluid with the Taguchi predicted optimal set of parameters has been performed and OYS matches well (a marginal error of 1.54%) with its statistically predicted value. The ANOVA results examine the contribution of input parameters and present that the iron particle proportion is the most prominent parameter that impacts the OYS, contributing 76.36%, followed by iron particle type which contributes 19.75%, carrier fluid with a contribution of 3.11% and additive proportion has a contribution of less than 0.1%. The OYS values for the optimal MR fluid are compared with commercially available MR fluids i.e., Lord MRF-122EG and MRF-132DG at various magnetic field intensities (0 - 164 kA/m). The comparison results exhibit that the OYS for the optimal in-house prepared MR fluid follows the same pattern as followed by MRF-122EG and MRF-132DG at all values of applied magnetic fields.

Intracellular doppler spectroscopy of live tissue sentinels for a fast in-vitro bacterial infection assay

Living organisms are frequently used as sentinels to monitor changes in the environment, and organoids can serve a similar function as in vitro sentinels detecting microbiological processes. One such process of great interest is pathogenic infection, which causes alterations in the cellular dynamics of the sentinels. Doppler light scattering is well suited to measure such changes, as it is sensitive to the intracellular dynamics of the organoids even when the microbe density is too dilute to detect directly. In this paper, we present measurements of the endogenous changes of intracellular dynamics of DLD organoids (microclusters) in response to bacterial infection, detected using a long-coherence form of biodynamic imaging (BDI), a dynamic-contrast digital holography technique that captures the Doppler shifts encoded in the dynamic speckles. With a stable common-path digital holography system and using small in-vitro tissue microclusters, clinically relevant inoculum doses as low as 100 cfu/mL cause detectable changes in intracellular Doppler spectra within hours of infection and several hours before the proliferating bacteria population causes direct light scattering. Because it can measure effects for small bacterial populations, the method shows promise as a fast diagnostic tool to detect the presence of bacteria in patient fluid samples faster than traditional methods.

Phenotypic Characterization of Multidrug Resistant Klebsiella Spp. Isolated from Pediatric Patients

infections are difficult to eradicate due to the presence of β-lactamases. The current study aimed to identify multidrug resistant isolates in children due to their high prevalence in Lahore, Pakistan. Methodology. In this study, 150 samples of blood, urine, pus, ascitic fluid, pleural fluid, Bronchoalveolar lavage (BAL), and CSF were collected from patients admitted in the University of Child Health Sciences, The Children’s Hospital, Lahore from November 2021 to April 2022. Then, different microbiological techniques including culturing, Gram staining, identification of different species by using API, antibiotic susceptibility testing as well as molecular techniques including PCR, were used to identify MDR Klebsiella spp. positive samples. Results. Out of the 150 samples, 58 were found positive for Klebsiella spp. while 92 were negative for Klebsiella spp. Among these 58 strains, 48 came from urine samples, 8 from pus samples, and 2 from blood samples. Furthermore, 34 were collected from male and 24 from female patients. The age group of the sampled children was 6 to 10 years. Afterwards, these 58 isolates were screened for extended spectrum β-lactamase (ESBL), metallo β-lactamase (MBL) and AmpC. It was found that out of these 58 isolates, 12 were ESBL positive (20.68%), 11 were MBL positive (18.9%) and 20 were AmpC positive (34.48%). Among β-lactamase genes, NDM -1 (72.7 %), SHV (41.6%), TEM (8.3%) and CTX-M (25%) were detected. Conclusion. The study concluded that the rate of MDR Klebsiella infection is high in children and it may be complicated for the clinicians to treat them.

Host biliary bacteria in cholecystectomy at a secondary-care teaching hospital, an observational study.

Host biliary bacteria in cholecystectomy at a secondary-care teaching hospital, an observational study.

A Prospective Evaluation of the Diagnostic Utility for Low-Coverage Genome Sequencing in Prenatal Samples: A Comparison With Chromosomal Microarray Analysis.

The present study aimed to evaluate the efficacy of LC-GS in detecting clinically relevant chromosomal abnormalities in comparison with conventional CMA within a prenatal context. We conducted a prospective study involving 200 amniotic fluid samples. All specimens were analyzed via LC-GS and traditional tests, including CMA and karyotyping. LC-GS was performed with 3X coverage to gauge its proficiency in identifying copy number variations (CNVs), aneuploidies, regions of homozygosity (ROH), and chromosome mosaicism. Data from both methods were compared to evaluate their sensitivity, specificity, and overall clinical utility. LC-GS at a depth of 3X identified a total of 77 positive samples, yielding a detection rate of 38.5% (77/200). This included 17 cases of aneuploidy, 36 instances of CNVs, 20 cases linked to ROH, and 8 cases of chromosomal mosaicism. LC-GS demonstrated high concordance with CMA in aneuploidy, CNVs, ROH, and chromosomal mosaicism, achieving a diagnostic yield of 21% (42/200), with additional benefits of reduced cost. Moreover, LC-GS outperformed CMA in terms of resolution for identifying submicroscopic CNVs. LC-GS presents a robust alternative to CMA for prenatal diagnosis, effectively identifying aneuploidies, CNVs, chromosomal mosaicism, and ROH. It delivers comparable sensitivity and specificity in the detection of a wide spectrum of genomic abnormalities.