Fluid Samples Research Articles

Analysis of heat transfer coefficient in radiator cooling system using TiO2/CuO hybrid fluid

The automotive industry is growing, encouraging more efficient car cooling systems, especially radiators. Innovations are constantly being made to improve the performance of radiators. Choosing the correct fluid is one of the ways to increase heat transfer. This study aimed to compare the performance of coolant OBC and TiO2/CuO hybrid fluid mixed with coolant OBC to investigate the increase in heat transfer in the car's radiator. The first step is to make TiO2/CuO hybrid fluid with a two-step method, with volume fraction variations of 1.0, 2.0, 3.0, 4.0, and 5.0%, and observe stability for 30 days. Viscosity and thermal conductivity tests are used to assess the thermophysical properties of the sample. Hybrid fluid samples of TiO2 / CuO with a volume fraction of 5.0% showed the best stability and thermal conductivity, then were added to the coolant brand OBC (1:4). The results showed an increase in heat transfer rate of 23% and a heat transfer coefficient of 20% in the TiO2/CuO hybrid fluid added to the coolant OBC

Prevalence and Genetic Variation Investigation of the Pseudorabies Virus in Southwest China

In 2022, a significant PRV outbreak in a southwestern China pig farm led to a high incidence of sow abortion. A serological analysis using gE antigen-based ELISA revealed a high prevalence (69.30%) of PRV gE antibodies among the affected pigs, with a significant variation across different pig populations (1.11–76.12%). We collected additional 5552 pig serum samples and 580 pig cerebrospinal fluid (CSF) samples from various pig farms in Southwest China between 2022 and 2024. The seropositive rates for PRV gE antibodies ranged from 2.36% and 8.65% in the serum samples, while the positive detection rates for the PRV gE gene in the cerebrospinal fluid samples, as determined by PCR, were between 1.06% and 2.36%. The PCR analysis and sequencing of the PRV gB, gC, gE, and TK genes from eight randomly selected samples identified two distinct strains, CQ1 and CQ2. CQ1’s gC gene showed similarity to the vaccine strain Bartha, while the other genes aligned with Chinese classical strains, suggesting its potential genetic recombination. CQ2 aligned with the Chinese classical strain SC. Although the overall PRV infection in Southwest China’s pig farms is relatively low, occasional outbreaks with high positivity rates are observed. These findings highlight the necessity for increased surveillance and stringent control measures to safeguard the swine industry.

Coarse-Grained MD Simulations of the Capillary Interaction between a Sphere and a Binary Fluid with Truncated Lennard-Jones Potentials.

In atomic force microscopy experiments on fluid samples, a capillary bridge forms between the tip and the fluid, causing an attractive capillary force. Here, we present a computational model of the capillary interaction between a solid sphere and a coarse-grained Lennard-Jones fluid containing 10% antifreeze particles with an enlarged van der Waals radius. The capillary force acting on the sphere is obtained from the displacement of the sphere in a trap potential as the sphere is incrementally approached and then retracted from the fluid. This yields force-distance data similar to that obtained in atomic force microscopy experiments. We use this methodology to study the influence of the cutoff radius of the truncated Lennard-Jones potentials on the capillary force and its temperature dependence. The latter is found to scale with the critical temperature of the system. With the presented approach, the tip-sample interaction can be studied for a wide range of complex fluids, particle shapes, and force-probing schemes.

Changes in heavy metal levels in the oral fluid of young patients with and without plaque-induced gingivitis undergoing treatment with braces

Relevance. Research into the effects of nickel-titanium alloys and heavy metal ions on the development of inflammatory oral diseases is becoming increasingly significant in orthodontics. The relationship between toxic elements and inflammatory responses remains under-researched and controversial. Data on the connection between heavy metals and inflammatory reactions could influence the choice of orthodontic appliance materials and guide the subsequent management and monitoring of patients with braces. The question of the safe use of nickel-titanium alloy braces continues to be a relevant concern.Purpose. To assess the levels of heavy metals in the oral fluid of young patients with braces, both in those who developed plaque-induced gingivitis and those who did not.Materials and Methods. The study involved 50 clinically healthy patients (Groups I and II health categories) with intact teeth, including patients with compensated dental caries (isolated carious lesions – Grade I) and no periodontal diseases (PMA index <20), presenting with dental crowding and malocclusion (K07.2, K07.3). The average age of the patients was 22.1 ± 2.7 years, all of whom were undergoing treatment with metal braces. Oral fluid samples were analyzed for 12 heavy metal elements using inductively coupled plasma mass spectrometry (ICP-MS) in a laboratory setting. Additionally, lysozyme activity and oral fluid pH were measured before appliance placement and six months into orthodontic treatment.Results. Adaptation to braces in young patients is frequently complicated by the development of gingivitis and periodontitis due to impaired oral hygiene and the mechanical pressure exerted on teeth and gums, leading to elemental imbalances. Patients with gingivitis during orthodontic treatment are particularly at risk of developing such imbalances and therefore require timely correction of their mineral metabolism.

Comparative analysis of compartment-specific immunothrombotic biomarker profiles in bronchoalveolar lavage fluid and serum of patients with pneumonia-related acute respiratory distress syndrome: A preliminary cross-sectional study.

Immunothrombosis has emerged as a potential mechanistic link underlying the development and progression of acute respiratory distress syndrome (ARDS), but understanding its specific profile in patients, both locally and systemically, is limited. The objective of this study was to characterize and compare the immunothrombotic signatures in patients diagnosed with pneumonia-related ARDS (p-ARDS) at both the pulmonary and systemic levels and to evaluate their clinical relevance. The study included 23 consecutive patients diagnosed with p-ARDS admitted to the intensive care unit at a tertiary university hospital from July 2022 to May 2023, alongside 40 concurrently hospitalized patients with common pneumonia as controls. Paired bronchoalveolar lavage fluid (BALF) and serum samples were collected from the participants for the analysis of 15 biomarkers to assess and quantify the pulmonary and systemic immunothrombotic signatures. The study results revealed significant pulmonary inflammation and systemic endothelial injury in p-ARDS patients compared to pneumonia controls. These observations were maintained after adjustment for severity of illness (Acute Physiology and Chronic Health Evaluation II scores). In terms of clinical relevance, inflammatory biomarkers (interleukin [IL]-6, IL-8) in BALF were found to correlate with PaO2/FiO2 ratio, while serum levels of a disintegrin and metalloproteinase with thrombospondin type 1 motif 13 (ADAMTS-13) and thrombomodulin showed associations with Sequential Organ Failure Assessment and Disseminated Intravascular Coagulation scores. In conclusion, this preliminary investigation identified compartment-specific variations in the immunothrombotic signature between patients with p-ARDS and those with pneumonia alone, with inflammatory responses predominantly localized in the alveolar compartments and coagulation/endothelial injury biomarkers more pronounced in peripheral blood.

Intrathecal IgG and IgM synthesis correlates with neurodegeneration markers and corresponds to meningeal B cell presence in MS

Intrathecal synthesis of immunoglobulins (Igs) is a key hallmark of multiple sclerosis (MS). B cells are known to accumulate in the leptomeninges of MS patients and associate with pathology in the underlying cortex and a more severe disease course. However, the role of locally produced antibodies in MS brain pathology is poorly understood. Here, we quantified the protein levels of IgA, IgM, IgG and albumin in serum and cerebrospinal fluid (CSF) samples of 80 MS patients and 28 neurological controls to calculate Ig indices. In addition, we quantified presence of meningeal IgA+, IgM+ and IgG+ B cells in post-mortem brain tissue of 20 MS patients and 6 controls using immunostainings. IgM and IgG, but not IgA, indices were increased in CSF of MS patients compared to controls, with no observed differences between MS disease types. Both IgM and IgG indices correlated significantly with neurofilament light (NfL) levels in CSF, but not with clinical or radiological parameters of disease. Similarly, IgG+ and IgM+ B cells were increased in MS meninges compared to controls, whereas IgA+ B cells were not. Neuronal loss did not differ between sections with low or high IgA+, IgM+ and IgG+ B cells, but was increased in sections with high numbers of all CD19+ meningeal B cells. Similarly, high presence of CD19+ meningeal B cells and IgG+ meningeal B cells associated with increased microglial density in the underlying cortex. Taken together, intrathecal synthesis of IgG and IgM is elevated in MS, which corresponds to an increased number of IgG+ and IgM+ B cells in MS meninges. The significant correlation between intrathecal IgG and IgM production and NfL levels, and increased microglial activation in cortical areas adjacent to meningeal infiltrates with high levels of IgG+ B cells indicate a role for intrathecal IgM- and IgG-producing B cells in neuroinflammatory and degenerative processes in MS.

Comparison of DNA extraction procedures for detection of Mycoplasma bovis directly from extended bovine semen straw samples using a commercial M. bovis PCR

BackgroundMycoplasma bovis is a global pathogen of cattle but was detected for the first time in New Zealand in 2017, triggering a response under their Biosecurity Act as an “unwanted organism”. Following a lengthy eradication campaign, the Ministry of Primary Industries (MPI) now requires all bovine semen destined for export to New Zealand to be screened with an M. bovis-specific real-time PCR (rtPCR) compliant with amended import health standard (IHS) test requirements aimed at preventing the accidental importation of M. bovis. The standard stipulates that semen samples cannot be centrifuged prior to DNA extraction. To comply with these strict requirements, one of the listed tests was validated together with different DNA preparation steps and compared with existing in-house procedures. DNA was extracted from semen straws using the current in-house semi-automated platform procedures for processing culture, tissue and body fluid sample submissions and was compared with the stipulated test requirements. DNA from centrifuged and unspun semen samples spiked with M. bovis was also compared.ResultsThe rtPCR had a sensitivity and specificity of 100% (95% confidence interval = 79–100% and 74–100%, respectively) when testing DNA from other Mycoplasma species or bovine semen spiked with the latter, with a high level of repeatability for within- and between- run replicates. The consistent limit of detection was 0.001 pg/µl M. bovis DNA and between 5.3 × 102 and 7.5 × 102 CFU/ml M. bovis when artificially spiked in semen. DNA extracted using the KingFisher Flex was detected with lower Cq values than the Maxwell 16, but the comparable improvements in sensitivity were mainly associated with non-centrifuged samples (p < 0.001). None of the procedures tested impeded the detection sensitivity of M. bovis in the presence of competitor organisms Acholeplasma laidlawii, Mycoplasma bovigenitalium and Ureaplasma diversum, confirming M. bovis specificity of the polC target.ConclusionsUnder the experimental conditions applied, this rtPCR test efficiently detected M. bovis in extended bovine semen straw samples from DNA extracted using both semi-automated extraction platforms, irrespective of prior centrifugation of extended semen.

Fluid inclusions in magmatic ilmenite record degassing in basic magmas

Magmatic volatile phases play a major role in igneous systems, but indirect sampling of the magmatic fluid, especially for basic magmas, remains challenging to document. Here, we report compelling evidence of primary fluid inclusions trapped within magmatic ilmenite from two different basic intrusive settings: Armorican Massif (France) and Central Iberian Zone (Spain). Fluid inclusions have a solute chemistry dominated by sodium, calcium, chlorine, sulfur and iron, with detectable contents of metals and metalloids and thus likely record the onset of late-stage magmatic volatile saturation in these basic magmas. Hence, we argue the presence of fluid inclusions in ilmenite may be a good indicator for magma degassing in these settings and importantly records the magmatic-hydrothermal transition. Additionally, this study suggests that the trapping of the magmatic volatile phase during ilmenite (and other opaque minerals) crystallization may be more common but at present an underappreciated phenomenon in basic igneous magmatic systems.

Programming rumen microbiome development in calves with the anti-methanogenic compound 3-NOP

The aim of this study was to establish a distinctive rumen microbial and fermentation profile using the anti-methanogenic compound 3-NOP to assess dam effect, and nutritional intervention of the juvenile offspring on microbial structure and function of rumen up to 12 months of age, once the treatment was withdrawn. Forty-eight pregnant heifers (H) and their future offspring (C) were allocated to either Control (-) or 3-NOP (+) treatment resulting in four experimental groups: H+/C+, H+/C-, H-/C + and H-/C-. Animals were treated from 6 weeks prior to calving until weaning, with the offspring monitored until 12 months of age. Rumen fluid samples and methane measurements using the Greenfeed system were collected during the trial. Results supported the mode of action of the compound, with a shift in fermentation from acetate to propionate, increases in branched chain fatty acids and formic acid in the 3-NOP treated animals. Similar shifts in microbial populations occurred in 3-NOP treated animals with lower abundances of rumen methanogen populations, increases of bacterial groups Succiniclasticum spp, Candidatus Saccharimonas. Fibrobacter and the families Prevotellaceae and Succinivibrioacea. and the protozoa Entodinium. Early life intervention had an enduring impact on the rumen microbial structure of young animals up to 28 weeks post weaning, however the effect was diminished once 3-NOP was withdrawn. Interestingly, a group of young animals emitted significantly less methane (15%) than the animals that did not receive the treatment during their juvenile stage. Our results suggest a higher resemblance of the young calf microbiome to a low methane adult and that early life colonisation of the rumen persists through to later life with the pre-weaning microbiome comprising ~ 65% of the yearling animal. Further research needs to be performed to determine the timing and dose of 3-NOP for new-born calves that can sustain a reduction in methane emissions after the treatment is withdrawn, under extensive grazing or controlled conditions.

Impact of Long-term Fasting on Breath Volatile Sulphur Compounds, Inflammatory Markers and Saliva Microbiota Composition.

Despite substantial evidence supporting the role of resident bacterial communities in therapeutic fasting outcomes, research has primarily focused on gut microbiota, leaving changes in oral microbiota largely unexplored. The clinical significance of oral health changes during fasting is nonetheless underscored by the documented development of halitosis in fasting individuals. However, no scientific studies have comprehensively examined the interplay between salivary microbiota alterations, inflammatory changes in the gingival crevice, and the production of malodorous volatile compounds. We examined volatile sulphur compounds (VSC) in breath during fasting, cytokine levels in the gingival crevice, and oral microbiota composition of the saliva in a single-arm interventional study involving 36 subjects who fasted for 10 ± 3 days. Participants fasted according to Buchinger fasting guidelines. VSC were evaluated every morning before any food or drink intake using the OralChroma gas chromatography device. Saliva and gingival crevicular fluid (GCF) samples were collected at the clinical site before fasting, at the end of fasting, and at the end of food reintroduction. Follow-up saliva samples were sent to the patients after 1 and 3 months. Saliva samples were processed and analysed by targeted sequencing of 16S rRNA gene amplicons, whereas the expression of 6 inflammatory markers in the GCF were analysed using a multiplex fluorescent bead-based immunoassay. The quantification of volatile compounds in the breath demonstrated a statistically significant increase in dimethylsulfide levels during fasting, which corroborates the occurrence of bad breath as a common side effect of fasting. Salivary microbiota profiling showed a shift in microbial composition, including reduction in the levels of Neisseria, Gemella and Porphyromonas spp., concomitant with an increase in the levels of Megasphaera, Dialister, Prevotella, Veillonella, Bifidobacteria, Leptotrichia, Selenomonas, Alloprevotella, and Atopobium. We further demonstrated a reduction in the levels of the pro-inflammatory cytokine interleukin-8 in the GCF. Dimethylsulfide concentrations in the breath increased during fasting, and this was correlated to changes in the oral microbiota. Future studies are needed to illuminate the possible impact of these changes on oral and general health status.

Transplantation of human umbilical cord-derived mesenchymal stem cells improves age-related ovarian functional decline via regulating the local renin–angiotensin system on inflammation and oxidative stress

BackgroundAge-related reproductive aging is a natural and irreversible physiological process, and delaying childbearing is increasingly common all over the world. Transplantation of mesenchymal stem cells (MSCs) is considered a new and effective therapy to restore ovarian function, but the relevant mechanisms remain unclear. Recently, it has been found that there is a local Renin–angiotensin system (RAS) in human ovary and it plays a key role.MethodsAfter collecting follicular fluid from women who received oocyte retrieval for pure male factor infertility, the level of RAS components in it were detected, and the correlation analysis by linear regression. Then, the in vivo experiments on female C57BL/6 mice were designed to measure ovarian function, and the transcription and translation levels of RAS pathway were detected by molecular biology methods. Moreover, the role of RAS in regulating inflammation and oxidative stress in the co-culture system were explored in in vitro experiments on KGN cells.ResultsFirst, a total of 139 samples of analyzable follicular fluid were obtained. The local RAS of ovary, which is independent of systemic RAS (P > 0.05), is affected by age (Pearson r < 0, P < 0.05) and related to ovarian function, inflammation, oxidative stress indexes and assisted reproduction laboratory outcomes (P < 0.05). Next, the ovary/body weight of aging mice decreased significantly and serum sex hormones levels changed significantly (P < 0.01). The number of functional follicles decreased, while the atresia follicles increased (P < 0.05). After MSCs transplantation, all the above measures have been partially recovered (P < 0.05). Although several RAS components in aging ovary changed, MSCs only improved the expression level of AT1R (P < 0.05). Furthermore, the secretion ability and mitochondrial membrane potential of aging KGN cells decreased, while the intracellular ROS level and the aging cells ratio increased (P < 0.01). All the above measures have been partially recovered when co-cultured with MSCs (P < 0.05). After Ang(1–7) were added into the co-culture system, the above have been more significantly restored compared with Ang II (P < 0.05). Nevertheless, there was no statistical difference in estradiol level no matter which one was added (P > 0.05).ConclusionsTogether, our findings indicate that a novel possible mechanism to explain how stem cells restore age-related ovarian functional decline.

Non-tuberculous mycobacteria enhance the tryptophan-kynurenine pathway to induce immunosuppression and facilitate pulmonary colonization

The increasing prevalence of non-tuberculous mycobacterium (NTM) infections alongside tuberculosis (TB) underscores a pressing public health challenge. Yet, the mechanisms governing their infection within the lung remain poorly understood. Here, we integrate metagenomic sequencing, metabolomic sequencing, machine learning classifiers, SparCC, and MetOrigin methods to profile bronchoalveolar lavage fluid (BALF) samples from NTM/TB patients. Our aim is to unravel the intricate interplay between lung microbial communities and NTM/Mycobacterium tuberculosis infections. Our investigation reveals a discernible reduction in the compositional diversity of the lung microbiota and a diminished degree of mutual interaction concomitant with NTM/TB infections. Notably, NTM patients exhibit a distinct microbial community characterized by marked specialization and notable enrichment of Pseudomonas aeruginosa and Staphylococcus aureus, driving pronounced niche specialization for NTM infection. Simultaneously, these microbial shifts significantly disrupt tryptophan metabolism in NTM infection, leading to an elevation of kynurenine. Mycobacterium intracellulare, Mycobacterium paraintracellulare, Mycobacterium abscessus, and Pseudomonas aeruginosa have been implicated in the metabolic pathways associated with the conversion of indole to tryptophan via tryptophan synthase within NTM patients. Additionally, indoleamine-2,3-dioxygenase converts tryptophan into kynurenine, fostering an immunosuppressive milieu during NTM infection. This strategic modulation supports microbial persistence, enabling evasion from immune surveillance and perpetuating a protracted state of NTM infection. The elucidation of these nuanced microbial and metabolic dynamics provides a profound understanding of the intricate processes underlying NTM and TB infections, offering potential avenues for therapeutic intervention and management.

Effect of a Bacillus subtilis-based direct-fed microbial, on milk yield, milk components, feed intake and plasma hormones and metabolites in lactating Holstein cows

To investigate the effects of a Bacillus-based direct-fed microbial on milk production and related factors, a study was conducted on Holstein cows (n = 28) starting at 90 ± 11 DIM. The cows were divided into two dietary groups: a control group (CON) that received a total mixed ration (TMR), and a Bacillus-fed group (DFM) that received the same TMR along with a Bacillus subtilis product containing two strains (747 and 1781) in equal amounts. The study lasted for 25 wk and included both primiparous and multiparous cows. The cows were housed in a free-stall barn and were provided with ad libitum TMR, which was fed twice a day. Their daily meals and dry matter intake (DMI) were recorded during wk 1–4 and wk 19–25 using electronic feeders. Milk samples were collected weekly during morning and evening milkings and analyzed for milk fat, protein, lactose, and milk urea nitrogen (MUN). Blood samples were also collected weekly and analyzed for plasma glucose, insulin, cholesterol, insulin-like growth factor 1 (IGF1), and progesterone concentrations. Additionally, rumen fluid samples were collected and evaluated for various species of bacteria. One of the key findings was that daily 4 % fat-corrected milk production (FCM) was influenced by the interaction between treatment (DFM vs. CON) and parity (primiparous vs. multiparous), with multiparous DFM cows producing 11 % more FCM compared to CON. The DFM cows also had a higher milk fat percentage (4.41 % vs. 4.02 %) and MUN concentrations were 1.0 mg/dL higher in the DFM group. In terms of blood metabolites and hormones, multiparous DFM cows had lower plasma cholesterol, glucose, and insulin concentrations compared to CON multiparous cows. However, there were no significant differences between the groups in terms of plasma IGF1 concentrations, insulin-to-glucose ratios, and luteal phase plasma progesterone concentrations. The analysis of rumen fluid samples revealed that the abundance of Ruminococcus albus and Fibrobacter succinogenes group I was greater in the DFM cows compared to CON cows. Furthermore, during the periods when feed intake was measured, the DFM cows had a 9 % reduction in feed intake and a 14 % improvement in feed efficiency (FCM per DMI) compared to CON cows. In conclusion, the two-strain Bacillus product used in this study (Certillus) showed potential as an effective direct-fed microbial. It was found to increase feed efficiency and milk production by altering the composition of ruminal microbiota and metabolism in the cows.

Enhancing Th17 cells drainage through meningeal lymphatic vessels alleviate neuroinflammation after subarachnoid hemorrhage

BackgroundSubarachnoid hemorrhage (SAH) is a severe cerebrovascular disorder primarily caused by the rupture of aneurysm, which results in a high mortality rate and consequently imposes a significant burden on society. The occurrence of SAH initiates an immune response that further exacerbates brain damage. The acute inflammatory reaction subsequent to SAH plays a crucial role in determining the prognosis. Th17 cells, a subset of T cells, are related to the brain injury following SAH, and it is unclear how Th17 cells are cleared in the brain. Meningeal lymphatic vessels are a newly discovered intracranial fluid transport system that has been shown to drain large molecules and immune cells to deep cervical lymph nodes. There is limited understanding of the role of the meningeal lymphatic system in SAH. The objective of this research is to explore the impact and underlying mechanism of drainage Th17 cells by meningeal lymphatics on SAH.MethodsTreatments to manipulate meningeal lymphatic function and the CCR7-CCL21 pathway were administered, including laser ablation, injection of VEGF-C geneknockout, and protein injection. Mouse behavior was assessed using the balance beam experiment and the modified Garcia scoring system. Flow cytometry, enzyme-linked immunosorbent assays (ELISA), and immunofluorescence staining were used to study the impact of meningeal lymphatic on SAH drainage. Select patients with unruptured and ruptured aneurysms in our hospital as the control group and the SAH group, with 7 cases in each group. Peripheral blood and cerebrospinal fluid (CSF) samples were assessed by ELISA and flow cytometry.ResultsMice with SAH showed substantial behavioral abnormalities and brain damage in which immune cells accumulated in the brain. Laser ablation of the meningeal lymphatic system or knockout of the CCR7 gene leads to Th17 cell aggregation in the meninges, resulting in a decreased neurological function score and increased levels of inflammatory factors. Injection of VEGF-C or CCL21 protein promotes Th17 cell drainage to lymph nodes, an increased neurological function score, and decreased levels of inflammatory factors. Clinical blood and CSF results showed that inflammatory factors in SAH group were significantly increased. The number of Th17 cells in the SAH group was significantly higher than the control group. Clinical results confirmed Th17 cells aggravated the level of neuroinflammation after SAH.ConclusionThis study shows that improving the drainage of Th17 cells by meningeal lymphatics via the CCR7-CCL21 pathway can reduce brain damage and improve behavior in the SAH mouse model. This could lead to new treatment options for SAH.

Radio-Biological Confrontation in the Diagnosis of Invasive Aspergillosis: About Two Cases and Review of Literature

Invasive pulmonary aspergillosis due to Aspergillus fumigatus is an often serious mycosis, usually occurring in severely immunocompromised patients. It is accompanied by a high mortality rate due to its often difficult and late diagnosis. Diagnosis is based on early chest computed tomography and mycological examination of respiratory samples. Both patients had presented with acute respiratory distress, developing in a context of fever and deterioration of the general condition despite the initiation of broad-spectrum empirical treatment. In each of them, a chest CT scan was performed, objectifying atypical cavitary images not suggesting pulmonary aspergillosis. Cytobacteriological examination of sputum and search for Mycobacterium tuberculosis were negative. Serology for aspergillus antigens and a sample of bronchoalveolar lavage fluid for a mycological study were requested. Direct examination of bronchoalveolar lavage fluid was negative, but culture allowed the identification of Aspergillus fumigatus. Galactomannan testing was positive. Given these results, treatment with injectable voriconazole was initiated in our two cases. The evolution was marked by a clinical improvement of one patient and the death of the other patient. Invasive pulmonary aspergillosis is a major concern due to the frequency of its complications. Its diagnosis must be early to ensure adequate management for a better prognosis.

Effects of water extracts of Artemisia annua L. on rumen immune and antioxidative indexes, fermentation parameters and microbials diversity in lambs

The present study investigated the effects of water extracts of Artemisia annua L. (WEAA) on rumen immune and antioxidative indexes, fermentation parameters and microbial diversity in lambs. A total of 32 3-month-old Dorper × Han female lambs having comparable body weights (24±0.09 kg) were selected and were randomly assigned to four treatments, with eight repetitions for each treatment. The basal diet, consisting of 45% concentrate and 55% forage, was solely provided to the control group. For the other treatment groups, the basal diet was supplemented with WEAA at dosages of 500, 1000, and 1500 mg/kg diet, respectively. Rumen tissue samples were collected for the analysis of immune and antioxidative parameters, as well as related gene expression. Rumen fluid samples were collected to assess rumen fermentation parameters on days 30 and 60 and to evaluate the microbiota on day 60. Results showed that WEAA supplementation linearly or quadratically increased the content of sIgA, IL-4, IL-2 and the gene expression level of MyD88, IκB-α, IL-4, COX-2, iNOS in rumen tissue (p &amp;lt; 0.05), as well as the bacteria negatively associated with IL-6 (g_ [Eubacterium]_cellulosolvens_group). Furthermore, the addition of WEAA linearly or quadratically increased rumen T-SOD, GSH-Px (p &amp;lt; 0.05) and the gene expression level of Nrf2, SOD2, GSH-Px, HO-1 (p &amp;lt; 0.05), and decreased the rumen concentration of malondialdehyde (MDA) and gene expression level of Keap1 (p &amp;lt; 0.05), as well as the bacteria positively associated with T-AOC, T-SOD and GSH-Px (g_Lachnospiraceae_NK3A20_group, g_Saccharofermentans, g__Marvinbryantia, g_unclassified_f_Eggerthellaceae). The supplementation of WEAA caused the concentration of microprotein (MCP), total volatile fatty acids (TVFA), propionate to increase either linearly or quadratically, while reducing the concentration of NH3-N and the acetate/propionate ratio (A:P) in rumen fluid (p &amp;lt; 0.05). The addition of WEAA linearly or quadratically increased the abundance of Actinobacteriota, Cyanobacteria and Lachnospiraceae_NK3A20_group (p &amp;lt; 0.10), and g__Lachnospiraceae_NK3A20_group, g_Saccharofermentans, g_Marvinbryantia, g_Bifidobacterium were significantly abundant as specific microflora in the 1000 mg/kg WEAA supplementation group. In conclusion, dietary inclusion of 1000 mg/kg WEAA improved the rumen immune function, antioxidant status, rumen fermentation, and composition of rumen microbes in lambs.

Targeted next-generation sequencing - a promising approach in the diagnosis of Mycobacterium tuberculosis and drug resistance.

Targeted next-generation sequencing (tNGS) offers a high-throughput, culture-independent approach that delivers a comprehensive resistance profile in a significantly shorter turn-around time, making it promising in enhancing tuberculosis (TB) diagnosis and informing treatment decisions. This study aims to evaluate the performance of tNGS in the TB diagnosis and drug resistance detection of Mycobacterium tuberculosis (MTB) using MTB clinical isolates and bronchoalveolar lavage fluid (BALF) samples. A total of 143 MTB clinical isolates were assessed, tNGS, phenotypic antimicrobial susceptibility testing (AST), and AST based on whole genome sequencing (WGS) exhibited high concordance rates, averaging 95.10% and 97.05%. Among 158 BALF samples, culture, Xpert MTB/RIF, and tNGS reported 29, 70 and 111 positives, respectively. In the confirmed cases with etiological evidence (smears, cultures, or molecular test), the positive rate of tNGS (73/83, 87.95%) was higher than that of Xpert MTB (67/83, 80.72%). Additionally, 45% (27/60) of clinically diagnosed cases (with imaging or immunological evidence) were positive for tNGS. Further validation on the discrepant results between tNGS and Xpert MTB/RIF with droplet digital PCR (ddPCR) yielded 35 positives, tNGS detected all, and Xpert MTB/RIF only identified 6 positives. In conclusion, tNGS demonstrates robust and rapid performance in the identification of MTB and its associated drug resistance, and can be directly applied to clinical samples, positioning it as a promising approach for laboratory testing of tuberculosis.

Impact of Forage Sources on Ruminal Bacteriome and Carcass Traits in Hanwoo Steers During the Late Fattening Stages.

This study examined the effects of different forage sources on the ruminal bacteriome, growth performance, and carcass characteristics of Hanwoo steers during the fattening stage. In Korea, where high-concentrate feeding is common, selecting suitable forage is crucial for sustainable beef production. Fifteen 23-month-old Hanwoo steers, weighing an average of 679.27 ± 43.60 kg, were fed the following five different forage sources: oat hay (OAT), rye silage (RYE), Italian ryegrass (IRS), barley forage (BAR), and rice straw silage (RSS), alongside 1.5 kg of dry matter concentrate daily for five months. Carcass traits were evaluated post-slaughter, and rumen fluid samples were analyzed using full-length 16S rRNA gene sequencing to determine the bacteriome composition. The forage source significantly affected the alpha-diversity indices and bacteriome biomarkers linked to the feed efficiency and ruminal fermentation. Differences in the backfat thickness and meat yield index were noted, with alpha-diversity indices correlating with carcass traits. The phylum Planctomycetota, especially the family Thermoguttaceae, was linked to nitrogen fixation in high-protein diets like IRS, while the genus Limimorpha emerged as a biomarker for the meat yield. These findings highlight the importance of forage selection during late fattening to optimize beef production, considering diet and bacteriome shifts.

P27.24.A SYSTEMIC AND LOCAL THERAPIES BEFORE AND AFTER DIAGNOSIS OF LEPTOMENINGEAL DISEASE IN PATIENTS WITH BRAIN METASTASES: AN EXPLORATIVE, SINGLE-CENTER RETROSPECTIVE COHORT STUDY

Abstract BACKGROUND Leptomeningeal metastasis (LMD), the dissemination of cancer cells into leptomeninges and/or cerebrospinal fluid, is believed to increase in incidence due to improved survival of oncological patients. Real-world data on therapies before and after diagnosis of LMD in brain metastasis patients are still limited. We aimed to characterize local and systemic therapies before and after diagnosis of LMD in these patients. MATERIAL AND METHODS We included 120 patients (treated from 2012 to 2023) with underlying solid malignancy, presence of brain metastasis together with leptomeningeal spread, either confirmed by MRI, cerebrospinal fluid sampling, or both modalities. Data on demographics, radiological, and therapy-related characteristics were collected. Kaplan-Meier estimates and Cox model regression analysis were performed to identify factors associated with survival. RESULTS Breast cancer (35.8%), NSCLC (20.8%), melanomas (16.7%), and gastric cancer (5.7%) were among the most common entities. In 56% of cases, the diagnosis was made by MRI, in 5% by cerebrospinal fluid puncture (LP) alone, and in 39% by both MRI and LP. The median overall survival (OS) of the cohort was 2.77 months [95% CI: 2.27 - 4.20]. Patients who received systemic after diagnosis of LMD, either chemotherapy (n=26), targeted therapies (n=21) or checkpoint inhibitors (n=9) showed increase survival as compared to respective control patients. The only independent prognostic factors at diagnosis of LMD were presence of lung cancer (HR: 2.36 [95% CI: 1.28 - 4.4], p=0.006) and melanoma (HR: 2.30 [95% CI: 1.27 - 4.2], p=0.006). Local or systemic treatment modalities before LMD diagnosis were not associated with survival. CONCLUSION This study provides insights into the complex treatment modalities for LMD patients, both in terms of prior therapies and therapies after diagnosis of LMD. These results underscore the need for large prospective multi-center registries and interventional studies to further improve interdisciplinary and personalized therapy for these patients.

Torque teno virus (TTV) Infection in Patients with Encephalitis

Torque teno virus (TTV) is a ssDNA orphan virus belonging to the Anelloviridae family, but some recent studies suggested its possible involvement in central nervous system (CNS) pathology. We analyzed serum and cerebrospinal fluid samples (CSF) from 109 patients with encephalitis for TTV infection using serological and molecular testing, virus quantitative measurement, and next-generation sequencing-based (NGS) phylogenetic analysis. TTV noncoding region (UTR) and/or open reading frame 1 (ORF-1) sequences were detected in serum of 86 (79%) patients and in nine (8%) patients in CSF. Five of the latter patients were coinfected with various entero- and herpesviruses. Anti-TTV-IgG were detected in 80 (73.4%) sera and in two (1.8%) CSF samples, while anti-TTV-IgM were present in three (2.8%) sera and in none of the CSFs. Phylogenic analysis of CSF-derived TTV ORF-1 sequences revealed the presence of three unique variants in one patient. TTV was quantified in five CSF-serum pairs: in two patients viral loads were similar, and in three serum TTV loads were approximately one log higher. Our results suggest at least an occasional replication of TTV in CNS. However, whether TTV could be the cause of encephalitis requires further studies.