Sampling Time Points Research Articles

Influence of plastic shape on interim fragmentation of compostable materials during composting

Common experience with rotting wooden buildings demonstrates that fragmentation is a necessary natural process during biodegradation. In analogy, the loss of structural integrity of biodegradable plastics during biodegradation produces interim microplastic fragments. It is currently not known which parameters govern fragmentation kinetics: chemical structure, physical shape, and composite layers, or composting conditions may all be relevant. Here we investigated the influence of physical shape on the fragmentation of a polyester blend during laboratory tests simulating industrial composting. Methods previously validated on micronized granules as model shape were applied to shapes that better represent consumer products, such as micronized thin films and shredded plastic-coated paper cups. The peak interim number of detected fragments, which are between 3 and 2000 µm, ranked highest for micronized films, lower for micronized plastic granules, and even lower for coated paper cups. The layered structure of polyester on cellulose may thus have stabilized the biodegrading polyester compound against fragmentation. For thin films, fragment counts dissipated with halftime of 2.5 days, and less than 10–8% of the initially added polyester mass was detected in fragments between 3 and 25 µm at the last sampling time point. The physical shape and multilayer structure of the polymer-containing product were found to be decisive for fragmentation kinetics, indicating that tests on micronized polymer granules might not be representative of the release mechanism of fragments from consumer products containing plastic coatings.

Differentiation of bacterial communities on five common plastics after six days of exposure to Caribbean coastal waters

Plastic pollution in coastal areas, particularly in subtropical and tropical regions, remains a pervasive environmental issue. Marine plastic debris provides an artificial surface that rapidly accumulates a dynamic microbial biofilm upon entering the marine ecosystem. Especially the early stages of colonization are critical in shaping the microbial community. This study investigates the early microbial colonization, in less than a week, on five different plastic polymers in Caribbean coastal waters through 16S rRNA gene amplicon sequencing. We discovered shared bacterial taxa among the various plastic polymers and sampling timepoints, with dominant orders being Flavobacteriales, Rhodobacterales, Rhizobiales, and Pseudomonadales. Statistical analysis confirmed significant differences in community composition between the two sampling points, with polystyrene exhibiting a distinct microbial community on day 6 compared to polyethylene, polypropylene, and nylon. We found the same for polyethylene compared to nylon and polyethylene-terephthalate. Further examination identified 47 genera responsible for these differences, primarily belonging to the phyla Proteobacteria and Bacteroidota. Our data indicate an influence of both environmentally related stochastic processes and plastic-related specific factors during early colonization. Interestingly, we noticed an increase in the relative abundance of hydrocarbon and potentially plastic-degrading bacteria (PDB) from 12.4 to 34.5 % between the first and sixth day, suggesting their vital role in shaping the epiplastic community. Notably, some identified PDB have been reported to degrade the specific polymers studied, thus the monitored increase in relative abundance supports their role in plastic degradation. However, more research is required to fully understand their functioning and potential role in the epiplastic community. Our study provides insights into the prokaryotic colonization of marine plastics in the Caribbean basin, where to date studies have been limited despite high pollution rates.

Effect of Sampling Time, Quantification Method, and Tree Sample Pooling on Phytophthora cinnamomi Root Quantities in South African Avocado Orchards.

Phytophthora cinnamomi is a destructive soilborne pathogen causing Phytophthora root rot on avocados worldwide. Little is known about the effect of root sampling time, root quantification method (quantitative real-time PCR [qPCR] versus baiting), and tree sample pooling strategies on the quantification of the pathogen in roots in avocado orchard trees. This was investigated in six avocado orchards in two climatically different production regions (Mooketsi and Letaba) in the Limpopo Province, South Africa, over a 2-year period. Two different tree sample pooling strategies, consisting of either a four-pooled group (four groups each containing five pooled trees) or a single-pooled group (20 trees pooled) per 1 ha, were both shown to be suitable for quantifying P. cinnamomi in tree roots using qPCR or root baiting. P. cinnamomi root quantities from the two tree sample pooling strategies were significantly correlated for both quantification methods. Both quantification methods were suitable for quantifying the pathogen in roots, although qPCR was superior to root baiting at identifying significant differences in P. cinnamomi quantities among root sampling time points. The effect of sampling time was dependent on the investigated year. In 2017, root quantities, which were only evaluated using qPCR, did not reveal a consistent trend of a specific sampling time yielding the highest root quantities for most of the orchards. However, five of the orchards in 2018, based on the qPCR analyses, contained significantly higher P. cinnamomi root quantities in May (late autumn) than in March (early autumn), August (late winter), and October/November (late spring). In 2018, P. cinnamomi root DNA quantities were significantly positively correlated with the number of soil temperature hours at 20 to 24 and 20 to 29°C 2 months preceding the root sampling dates and negatively correlated with the number of hours at 15 to 19°C 2 months preceding root sampling. Our study has identified P. cinnamomi root quantification methods and tree sample pooling strategies, which will be useful for understanding the biology of the pathogen and when disease management strategies should be in place.

Systemic interrogation of immune-oncology-related proteins in patients with locally advanced prostate cancer undergoing androgen deprivation and intensity-modulated radiotherapy

PurposeThe primary objective was to establish whether blood-based leucine-rich alpha-2-glycoprotein (LRG1) can predict outcomes in patients with locally advanced prostate cancer undergoing androgen-deprivation therapy (ADT) and radiotherapy (RT) and to determine how it may relate to 92 immune-oncology (I-O)-related proteins in this setting.MethodsBaseline blood level of LRG1 from patients treated with ADT and RT enrolled in the CuPCa (n = 128) and IMRT (n = 81) studies was measured using ELISA. A longitudinal cohort with matched blood samples from start of ADT, start of RT, and end of RT protocol from 47 patients from the IMRT cohort was used to establish levels of I-O proteins by high-multiplexing Proximal Extension Assay by Olink Proteomics.Statistical analyses using Kaplan–Meier, Cox regression, and LIMMA analyses were applied to predict the prognostic value of LRG1 and its correlation to I-O proteins.ResultsHigh baseline levels of LRG1 predicted a low frequency of treatment failure in patients undergoing ADT + RT in both the CuPCa and the IMRT cohorts. LRG1 was moderately correlated with CD4, IL6, and CSF1. We identified I-O proteins predicting metastatic failure (MF) at different timepoints.ConclusionLRG1 biomarker is associated with I-O proteins and can be used to improve stratification and monitoring of prostate cancer patients undergoing ADT + RT. This work will require further in-depth analyses in independent cohorts with treatment outcome data.Graphical abstractStudy outline. A) Study cohorts. B) Sampling time points in a longitudinal cohort.

Screening for immune biomarkers associated with infection or protection against Ehrlichia ruminantium by RNA-sequencing analysis

Heartwater is one of the most economically important tick-borne fatal diseases of livestock. The disease is caused by the bacteria Ehrlichia ruminantium transmitted by Amblyomma ticks. Although there is evidence that interferon-gamma controls E. ruminantium growth and that cellular immune responses are protective, an effective recombinant vaccine for this disease is lacking. Analyses of markers associated with infection as well as protection will lead to a better understanding of the E. ruminantium immune response and corresponding pathways induced in sheep peripheral blood mononuclear cells (PBMC) will assist in development of such a vaccine. In this study, Biomarkers of infection (BMI) were identified as uniquely expressed genes during primary infection and biomarkers of protection (BMP) associated with immune to heartwater were identified post challenge. Sheep were experimentally infected and challenged with E. ruminantium infected ticks. The immune phenotypic and transcriptome profile of their PBMC were compared to their own naïve PBMC collected before infection. The study revealed 305 differentially expressed genes (DEGs) as BMI, of these 17 were upregulated at all three time-points investigated. These DEGs, form part of the bacterial invasion of epithelial cells Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway, and others detected from day 1 post infection and are considered predictive markers for early heartwater infection in ruminants. Similarly, a total of 332 DEGs were identified as BMP, of these 100 were upregulated and 75 were downregulated at all three time-points investigated. However, at D1PC most DEGs were downregulated (n = 1312) that correlated with a reduction in the % CD4 and CD8 T cells detected with flow cytometry. KEGG pathway analyses showed complete down regulation of T cell specific pathways possibly due to homing of immune cells to the site of infection after acquired immunity developed. At D4PC, expression levels of most of these downregulated genes increased and by D6PC they were upregulated. This indicates that the sampling time-point for biomarker analyses is important when results for acquired immune responses are inferred. This data identified DEGs that could be considered as biomarkers of protective immunity that can be used for identification of vaccine antigens and provides a strong foundation to further development of heartwater recombinant vaccines.

Effects of swimming exercise on neuropathic pain in a rat model: role of glutamate

ABSTRACT Objective The pain-reducing effects of the exercise were exerted through different mechanisms. Knowing more clear mechanisms helps to find more approach that is therapeutic. The objective of the present study is the evaluation of cerebrospinal fluid (CSF) glutamate level alteration in neuropathic pain rats and whether physical activity could modulate it. Methods In the present study 104 male Wistar rats weighing 180–220 g were randomly divided into 4 groups (Sham, Sham + Exe, Neuropathy, and Neuropathy + Exe) which in turn each group subdivided into 4 groups according to time points for behavioral testing and CSF sampling (Baseline, 2 weeks, 3 weeks, and 4 weeks). To induction of neuropathy (by chronic constriction injury,), after anesthetizing with a mixture of ketamine (80 mg/kg) and xylazine (10 mg/kg), the animal’s right sciatic nerve was exposed and was ligated using four movable catgut chromic suture 4/0. The exercise protocol included 25 min of daily swimming, 5 days a week for 4 weeks. Thermal hyperalgesia and mechanical tactile threshold were detected using the plantar test and Von Frey filaments, respectively. CSF glutamate level was determined using high-performance liquid chromatography. Results Findings indicated that mechanical and thermal thresholds significantly (p < 0.01, p < 0.05 respectively) decreased in the neuropathy group against that in sham groups. On the other hand, exercise significantly increased mechanical tactile threshold (p < 0.0012) and thermal threshold (p < 0.05) compared to the neuropathy group. Moreover, CSF glutamate level prominently (p < 0.01) was increased in the neuropathy group compared to the sham group, and swimming exercise significantly (p < 0.001) reduced it. In conclusion The present findings provide new evidence showing that medium-intensity swimming exercise attenuates pain-like behaviors in neuropathic pain animals, which is possibly due to decreasing CSF glutamate level and its neurotransmission.

Microbial and physicochemical water quality changes within distribution and premise plumbing systems during a chlorine conversion

A strategy for nitrification control within chloraminated drinking water systems (CDWSs) is to temporarily switch from chloramine secondary disinfection to free chlorine, also known as a free chlorine conversion (FCC). However, the long-term and beneficial effects of FCCs are unclear, especially regarding opportunistic pathogen occurrence. In this study, the impacts to microbial and physicochemical parameters were monitored throughout a CDWS implementing a FCC. Water samples were collected weekly for 4–6 weeks before, during, and after a FCC at eight locations: four distribution system and four residential sites. Monochloramine residual (mean±standard deviation) before and after the FCC averaged 1.8±0.9 and 1.6±1.0 parts per million (ppm) for all sites, respectively. Free chlorine levels averaged 2.3±0.9 ppm. There were no significant differences in turbidity and hardness at each location during the three time periods, but some were noted for pH, temperature, and orthophosphate levels across various sites and sampling periods. For all locations, heterotrophic plate count levels were lower during the FCC compared to the periods before and after. All samples from one residence were culture positive for P. aeruginosa which exhibited high levels before the FCC, decreasing levels during, and steadily increasing levels after. Additionally, one week prior to the FCC, sediment samples from two elevated storage tanks, ET-1 and ET-2, were analyzed with ET-1 displaying higher levels of culturable heterotrophic bacteria and molecularly detected total bacteria, Legionella spp., and nontuberculous mycobacteria (NTM), as well as presence of culturable P. aeruginosa and total coliforms compared to ET-2. Fourteen P. aeruginosa and total coliform isolates were whole genome sequenced with genetic differences observed depending on the sampling location and timepoint. Collectively, the observed differences in chemical and microbial parameters advocates for a better understanding of the effects associated with implementing FCCs to determine both their effectiveness and potential risks/rewards to water quality.

Mass spectroscopy reveals compositional differences in copepodamides from limnic and marine copepods

Marine copepods, the most abundant animals in the global ocean, imprint their surrounding waters with chemical cues, called copepodamides. Copepodamides induce defensive traits such as toxin production, bioluminescence, and colony size plasticity in a variety of marine phytoplankton. The role of copepodamides in freshwater ecosystems is, however, unknown. Here we report the consistent presence of copepodamides in copepods from six Swedish freshwater lakes. Copepodamide concentrations in freshwater copepods are similar to those of marine copepods, around 0.1 ppt of dry mass in millimetre sized individuals. The composition substantially overlaps with marine copepodamides but is also distinctly different. Marine copepods commonly contain both subgroups of copepodamides, the copepodamides (CA) and the dihydro-copepodamides (dhCA), whereas freshwater copepods are dominated by the dhCAs. Taxonomic groups had consistent copepodamide profiles across sampling sites and timepoints, supporting the presence of species-specific copepodamide signatures. We describe 10 new copepodamide structures, four of which were found exclusively in freshwater copepods. The presence of copepodamides in limnic copepods also warrants studies into their potential function as predator alarm cues in freshwater systems.

Impact of Applied Behavior Analysis on Autistic Children Target Behaviors: A Replication Using Repeated Measures.

Applied behavior analysis (ABA) is a primary evidence-based practice in treating autism spectrum disorder (ASD). Ongoing research is needed to report the resultsof ABArelative to attaining target behaviors. This study aims to replicate the results of previous research to determine the effectiveness of ABA of target behaviors in autisticchildren with a new timepoint sample of data. Materials & methods: A repeated measures analysis tracked 98 autistic children, which included four adult participants, over three timepoints during a one-month snapshot period from 6/7/23to 7/7/23. This study used a retrospective chart review to gather data on target behaviors to determine the effectiveness of ABA treatments across age categories. A mixed (between x within) analysis of variance (ANOVA) and subsequent post hoc and interaction contrasts were used to determine statistical significance. Mixed (between x within) ANOVAindicated statistical significance (sphericity assumed), F(2,160) = 32.893, and p < 0.05, across time. Using bootstrapped paired t-tests, multiple comparisons indicated p < 0.001 on all three multiple comparisons, with Bonferroni corrected α = 0.017. There was also a non-significant interaction effect (sphericity assumed) with (time) x (age category), F(8,160) = 0.333, p = 0.952, likely due to sizeable within-group variation resulting in a lowered statistical power. Conclusions: This replication found that autistic children receiving the ABA intervention demonstrated statistically significant improvement in target behaviors over the one-month snapshot period.

Feed supplementation with Ligilactobacillus salivarius PS21603 optimises intestinal morphology and gut microbiota composition in weaned piglets.

Post-weaning diarrhoea in piglets remains an important cause of economic losses for swine producers. Feed supplementation with probiotics is one of the alternatives to antibiotics used to reduce the impact of such gastrointestinal disease. The aim of the present study was to evaluate the effect of Ligilactobacillus salivarius PS21603 supplementation on the intestinal structure and the gut microbiota composition of weaned piglets. Safety and tolerance of L. salivarius PS21603 were previously evaluated in a 28-days study using 384 weaned piglets (28± 2 days old and 7.5± 1.5kg) divided in three treatment groups: T1: Basal diet+ L. salivarius PS21603 109 cfu/day, T2: Basal diet+ L. salivarius PS21603 107 cfu/day, and T3: Basal diet (control group). For the present study, 16 piglets per treatment group were randomly selected and faecal samples were collected on day 0 (weaning) and 28 of study. At the end of study, three males and three females per treatment were euthanised. Intestinal morphometric values were measured after necropsy. Faecal counts of Escherichia coli were evaluated by culture techniques, and faecal microbiota composition was assessed by high-throughput sequencing. All data were analysed and compared between treatment groups. Supplementation with L. salivarius PS21603 caused an increase in the intestine length of piglets from T1 and in the villous height:crypt ratio of piglets from T2 (P< 0.05) compared to T3 on day 28. According to the Shannon Diversity Index, microbiota diversity increased on day 28 compared to day 0, with no significant differences observed between treatments. The main changes in the relative abundance of bacteria at the phylum, family, and genus levels were observed between different sampling time points. However, piglets from T1 and T2 had lower faecal E. coli counts than T3 on day 28 (P< 0.05). Moreover, supplementation with L. salivarius PS21603 modulated gut microbiota through a more optimal composition, reducing Escherichia and increasing Bifidobacterium relative abundance in piglets from T1 (P< 0.05) from the beginning to the end of the study. Therefore, the strain L. salivarius PS21603 has shown probiotic properties to be used as feed additive in the pig industry, along with good hygiene and farm management practices, for the prevention and/or treatment of post-weaning diarrhoea in piglets.

Diet and monensin influence the temporal dynamics of the rumen microbiome in stocker and finishing cattle

BackgroundStocker cattle diet and management influence beef cattle performance during the finishing stage, but knowledge of the dynamics of the rumen microbiome associated with the host are lacking. A longitudinal study was conducted to determine how the feeding strategy from the stocker to the finishing stages of production affects the temporal dynamics of rumen microbiota. During the stocker phase, either dry hay or wheat pasture were provided, and three levels of monensin were administrated. All calves were then transported to a feedlot and received similar finishing diets with or without monensin. Rumen microbial samples were collected on d 0, 28, 85 during the stocker stage (S0, S28 and S85) and d 0, 14, 28, 56, 30 d before slaughter and the end of the trial during the finishing stage (F0, F14, F28, F56, Pre-Ba, and Final). The V4 region of the bacterial 16S rRNA gene of 263 rumen samples was sequenced.ResultsHigher alpha diversity, including the number of observed bacterial features and the Shannon index, was observed in the stocker phase compared to the finishing phase. The bacterial amplicon sequence variants (ASVs) differentiating different sampling time points were identified. Dietary treatments during the stocker stage temporally impact the dynamics of rumen microbiota. For example, shared bacteria, including Bacteroidales (ASV19) and Streptococcus infantarius (ASV94), were significantly higher in hay rumen on S28, S85, and F0, while Bacteroidaceae (ASV11) and Limivicinus (ASV15) were more abundant in wheat. Monensin affected rumen microbial composition at a specific time. Transportation to feedlot significantly influenced microbiome structure and diversity in hay-fed calves. Bacterial taxa associated with body weight were classified, and core microbiotas interacted with each other during the trial.ConclusionsIn summary, the temporal dynamics of the rumen microbiome in cattle at the stocker and finishing stage are influenced by multiple factors of the feeding strategy. Diet at the stocker phase may temporarily affect the microbial composition during this stage. Modulating the rumen microbiome in the steers at the stocker stage affects the microbial interactions and performance in the finishing stage.

Longitudinal Associations Between Persistent Post-Concussion Symptoms and Blood Biomarkers of Inflammation and CNS-Injury After Mild Traumatic Brain Injury.

The aim of our study was to investigate the biological underpinnings of persistent post-concussion symptoms (PPCS) at 3 months following mild traumatic brain injury (mTBI). Patients (n = 192, age 16-60 years) with mTBI, defined as Glasgow Coma Scale (GCS) score between 13 and 15, loss of consciousness (LOC) <30 min, and post-traumatic amnesia (PTA) <24 h were included. Blood samples were collected at admission (within 72 h), 2 weeks, and 3 months. Concentrations of blood biomarkers associated with central nervous system (CNS) damage (glial fibrillary acidic protein [GFAP], neurofilament light [NFL], and tau) and inflammation (interferon gamma [IFNγ], interleukin [IL]-8, eotaxin, macrophage inflammatory protein-1-beta [MIP]-1β, monocyte chemoattractant protein [MCP]-1, interferon-gamma-inducible protein [IP]-10, IL-17A, IL-9, tumor necrosis factor [TNF], basic fibroblast growth factor [FGF]-basic platelet-derived growth factor [PDGF], and IL-1 receptor antagonist [IL-1ra]) were obtained. Demographic and injury-related factors investigated were age, sex, GCS score, LOC, PTA duration, traumatic intracranial finding on magnetic resonance imaging (MRI; within 72 h), and extracranial injuries. Delta values, that is, time-point differences in biomarker concentrations between 2 weeks minus admission and 3 months minus admission, were also calculated. PPCS was assessed with the British Columbia Post-Concussion Symptom Inventory (BC-PSI). In single variable analyses, longer PTA duration and a higher proportion of intracranial findings on MRI were found in the PPCS group, but no single biomarker differentiated those with PPCS from those without. In multi-variable models, female sex, longer PTA duration, MRI findings, and lower GCS scores were associated with increased risk of PPCS. Inflammation markers, but not GFAP, NFL, or tau, were associated with PPCS. At admission, higher concentrations of IL-8 and IL-9 and lower concentrations of TNF, IL-17a, and MCP-1 were associated with greater likelihood of PPCS; at 2 weeks, higher IL-8 and lower IFNγ were associated with PPCS; at 3 months, higher PDGF was associated with PPCS. Higher delta values of PDGF, IL-17A, and FGF-basic at 2 weeks compared with admission, MCP-1 at 3 months compared with admission, and TNF at 2 weeks and 3 months compared with admission were associated with greater likelihood of PPCS. Higher IL-9 delta values at both time-point comparisons were negatively associated with PPCS. Discriminability of individual CNS-injury and inflammation biomarkers for PPCS was around chance level, whereas the optimal combination of biomarkers yielded areas under the curve (AUCs) between 0.62 and 0.73. We demonstrate a role of biological factors on PPCS, including both positive and negative effects of inflammation biomarkers that differed based on sampling time-point after mTBI. PPCS was associated more with acute inflammatory processes, rather than ongoing inflammation or CNS-injury biomarkers. However, the modest discriminative ability of the models suggests other factors are more important in the development of PPCS.

An Intravenous Pharmacokinetic Study of Cannabidiol Solutions in Piglets through the Application of a Validated Ultra-High-Pressure Liquid Chromatography Coupled to Tandem Mass Spectrometry Method for the Simultaneous Quantification of CBD and Its Carboxylated Metabolite in Plasma.

Cannabidiol (CBD) has multiple therapeutic benefits that need to be maximized by optimizing its bioavailability. Numerous formulations are therefore being developed and their pharmacokinetics need to be studied, requiring analytical methods and data from intravenous administration. As CBD is susceptible to hepatic metabolism, the requirement of any method is to quantify metabolites such as 7-COOH-CBD. We demonstrated that CBD and 7-COOH-CBD could be simultaneously and correctly quantified in piglet plasma by using an UHPLC-MS/MS technique. The validated method allowed for an accurate bioanalysis of an intravenously injected solution consisting of CBD-HPβCD complexes. The experimental pharmacokinetic profile of CBD showed multi-exponential decay characterized by a fast apparent distribution half-life (0.25 h) and an elimination half-life of two hours. The profile of 7-COOH-CBD was not linked with the first-pass metabolism, since 80% of the maximum metabolite concentration was reached at the first sampling time point, without any decrease during the period of study. A two-compartment model was optimal to describe the experimental CBD profile. This model allowed us to calculate macro-micro constants and volumes of distribution (Vss = 3260.35 ± 2286.66 mL) and clearance (1514.5 ± 261.16 mL·h-1), showing that CBD is rapidly distributed to peripheral tissues once injected and slowly released into the bloodstream.

Soluble terminal complement complex blood levels are elevated in schizophrenia

The role of the complement system in schizophrenia (Sz) is inconclusive due to heterogeneity of the disease and study designs. Here, we assessed the levels of complement activation products and functionality of the classical pathway in acutely ill unmedicated Sz patients at baseline and after 6 weeks of treatment versus matched controls. The study included analyses of the terminal complement complex (sTCC) and C5a in plasma from 96 patients and 96 controls by enzyme-linked immunosorbent assay. Sub-group analysis of serum was conducted for measurement of C4 component and activity of the classical pathway (28 and 24 cases per cohort, respectively). We found no differences in levels of C5a, C4 and classical pathway function in patients versus controls. Plasma sTCC was significantly higher in patients [486 (392–659) ng/mL, n = 96] compared to controls [389 (304–612) ng/mL, n = 96] (p = 0.027, δ = 0.185), but not associated with clinical symptom ratings or treatment. The differences in sTCC between Sz and controls were confirmed using an Aligned Rank Transformation model considering the covariates age and sex (p = 0.040). Additional analysis showed that sTCC was significantly associated with C-reactive protein (CRP; p = 0.006). These findings suggest that sTCC plays a role in Sz as a trait marker of non-specific chronic immune activation, as previously described for CRP. Future longitudinal analyses with more sampling time points from early recognition centres for psychoses may be helpful to better understand the temporal dynamics of innate immune system changes during psychosis development.

Temporal stability of fecal metabolomic profiles in irritable bowel syndrome.

The potential of the fecal metabolome to serve as a biomarker for irritable bowel syndrome (IBS) depends on its stability over time. Therefore, this study aimed to determine the temporal dynamics of the fecal metabolome, and the potential relationship with stool consistency, in patients with IBS and healthy subjects. Fecal samples were collected in two cohorts comprising patients with IBS and healthy subjects. For Cohort A, fecal samples collected during 5 consecutive days were analyzed by gas chromatography-tandem mass spectrometry (GC-MS/MS). For Cohort B, liquid chromatography-MS (LC-MS) was used to analyze fecal samples collected at week 0 (healthy and IBS) and at week 4 (patients only). Stool consistency was determined by the Bristol Stool Form scale. Fecal samples were collected from Cohort A (seven healthy subjects and eight IBS patients), and Cohort B (seven healthy subjects and 11 IBS patients). The fecal metabolome of IBS patients was stable short-term (Cohort A, 5 days and within the same day) and long-term (Cohort B, 4 weeks). A similar trend was observed over 5 days in the healthy subjects of Cohort A. The metabolome dissimilarity was larger between than within participants over time in both healthy subjects and IBS patients. Further analyses showed that patients had greater range of stool forms (types) than healthy subjects, with no apparent influence on metabolomic dynamics. The fecal metabolome is stable over time within IBS patients as well as healthy subjects. This supports the concept of a stable fecal metabolome in IBS despite fluctuations in stool consistency, and the use of single timepoint sampling to further explore how the fecal metabolome is related to IBS pathogenesis.

Grapevine Leaf Epiphytic Fungal and Bacterial Communities Are Influenced More by Spatial and Temporal Factors than by Powdery Mildew Fungicide Spray Programs

Synthetic fungicide treatments for managing grapevine powdery mildew have been widely applied in vineyards for decades, yet their ecological impacts on microorganisms in the phyllosphere have not been sufficiently studied. To investigate the impact of fungicide sprays on the grapevine leaf microbiota, grapevine leaf epiphytes were collected throughout the growing season from two California vineyards, each treated with identical spray programs. Each spray program was designed to control grapevine powdery mildew and contained five rotating synthetic fungicides. Leaf samples were collected before the first spray and between subsequent spray applications. Amplicon sequencing of ITS2 and 16S rRNA V4 regions was used to compare leaf epiphytic fungal and bacterial/archaeal diversities, taxonomic compositions, and differential abundances of taxa among treatments, locations, and sampling time points. Location (vineyard) and time significantly affected bacterial and fungal community compositions, whereas spray treatment did not. Plant-pathogenic fungi were the dominant fungal guild in both locations, and, in one location, their dominance increased significantly as the season progressed. Botrytis, an important postharvest pathogen in grapes, was significantly higher in relative abundance in one of the locations in the late season and acted as a hub genus in a co-occurrence network. In summary, the overall impact of the different spray programs on microbial community compositions was not significant, suggesting that the shifts of grapevine foliar microbial communities may be driven mainly by spatial and temporal factors rather than specific fungicide applications. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Frequency of PCV-2 viremia in nursery piglets from a Spanish swine integration system in 2020 and 2022 considering PRRSV infection status.

Porcine circovirus 2 (PCV-2) poses a significant economic threat for the swine industry, causing a range of diseases collectively referred to as porcine circovirus diseases (PCVDs). Despite PCV-2 vaccine effectiveness, the need for monitoring infectious pressure remains. PCV-2 coinfection with other pathogens like porcine reproductive and respiratory syndrome virus (PRRSV) can exacerbate disease severity and lead to PCV-2-systemic disease cases. Monitoring both PRRSV and PCV-2 in co-infected farms is crucial for an effective management and vaccination programs. The present cross-sectional study aimed to determine PCV-2 antibody levels in piglets at weaning and PCV-2 and PRRSV viremia in pooled serum samples at weaning (vaccination age) and at 6 and 9weeks of age from a Spanish swine integration system in 2020 (48 farms) and in 2022 (28 out of the 48 analysed previously). The frequency of PCV-2 detection in pools of piglet sera was 2.1% (2020) and 7.1% (2022) at vaccination age but increased at the end of the nursery period (10.4% in 2020 and 39.3% in 2022)in both years. Co-infections between PCV-2 and PRRSV were detected in a significant proportion of PRRSV positive farms (15% in 2020, and 60% in 2022). PCV-2 antibody levels(ELISA S/P ratios) at weaning were lower in PCV-2 qPCR positive farms at different sampling time-points (0.361 in 2020 and 0.378 in 2022) compared to PCV-2 qPCR negative ones (0.587 in 2020 and 0.541 in 2022). The 28 farms tested both years were classified in four different epidemiological scenarios depending on their PCV-2 virological status. Those PCV-2 qPCR negative farms in 2020 that turned to be positive in 2022 had a statistically significant increase of PRRSV RT-qPCR detection and a PCV-2 antibody levels reduction, facts that were not observed in the rest of the scenarios. This epidemiological study in farms from the same integration system determined the occurrence, in 2020 and in 2022, of PCV-2 and PRRSV infections in piglets during the nursery period by using pooled serum samples.

Soybean microbiome composition and the impact of host plant resistance.

Microbial communities play an important role in the growth and development of plants, including plant immunity and the decomposition of complex substances into absorbable nutrients. Hence, utilizing beneficial microbes becomes a promising strategy for the optimization of plant growth. The objective of this research was to explore the root bacterial profile across different soybean genotypes and the change in the microbial community under soybean cyst nematode (SCN) infection in greenhouse conditions using 16S rRNA sequencing. Soybean genotypes with soybean cyst nematode (SCN) susceptible and resistant phenotypes were grown under field and greenhouse conditions. Bulked soil, rhizosphere, and root samples were collected from each replicate. Sequencing of the bacterial 16S gene indicated that the bacterial profile of soybean root and soil samples partially overlapped but also contained different communities. The bacterial phyla Proteobacteria, Actinobacteria, and Bacteroidetes dominate the soybean root-enriched microbiota. The structure of bacteria was significantly affected by sample year (field) or time point (greenhouse). In addition, the host genotype had a small but significant effect on the diversity of the root microbiome under SCN pressure in the greenhouse test. These differences may potentially represent beneficial bacteria or secondary effects related to SCN resistance.

Design of Short-Term Power Load Forecasting Model Based on Deep Neural Network

In power system operation and planning, the accuracy of short-term power load forecasting is very important. Because of its powerful data processing and modeling ability, deep neural network has become an effective tool to accurately predict short-term power load. In this study, a short-term power load prediction model based on deep neural network is designed, which adopts deep long short-term memory and threshold period unit model, and combines Boosting algorithm for model fusion. The results show that the average absolute percentage error of the model fused by Boosting algorithm is 0.07%, which is 1.02% lower than the average weight method and 0.59% lower than the reciprocal error method. Boosting fusion model can effectively reduce the overall prediction error and maintain high stability of prediction error at peak, plateau and time sampling points, so as to achieve good prediction effect. Specifically, the MAPE of the model fused using Boosting algorithm is 0.07% (95% confidence), which is 1.14% higher than the average weight method and 0.79% higher than the reciprocal error method. The design of short-term power load forecasting model based on deep neural network can provide more accurate prediction for power system operation and planning, and help to improve the operation efficiency and reliability of power system. At the same time, the design and application of this model also provide a new idea and method for the application of deep learning in power system. The introduction of Boosting algorithm further improves the prediction accuracy and stability of the model, which is a major innovation in model design.

Exploring the effects of transport duration on the fecal microbial communities of surplus dairy calves

Transportation significantly affects the health and welfare of surplus dairy calves, largely due to the various stressors and pathogen exposures encountered during the process. Concurrently, an animal's microbiome is known to correlate with its health status, with stress-induced alterations in the microbiota potentially precipitating various diseases. This study aimed to compare the effects of transportation durations of 6, 12, or 16 h on the fecal microbiota in young surplus dairy calves. We used a randomized controlled design in which surplus dairy calves aged 1 to 19 d from 5 commercial dairy farms in Ontario were allocated into 1 of 3 transportation groups (6, 12, and 16 h of continuous transportation). Health assessments were conducted before, immediately after, and for 2 wk following transportation. Fecal samples were collected before, immediately after, and at 24 and 72 h after transportation and subjected to 16S rRNA sequencing. Alpha diversity metrics showed no significant differences between the 3 transportation groups at any of the sampling time points. Although β diversity metrics revealed no clustering by transportation groups, they indicated significant differences across sampling time points within each group. The overall analysis revealed a total of 22 phyla and 353 genera, with Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, and Fusobacteria being the most abundant phyla. Bacteroides, Escherichia/Shigella, Lactobacillus, Collinsella, and Bifidobacterium were the most abundant genera. The reduction in Fusobacteria abundance before and after transport was significantly larger in the 16-h transportation group when compared with the 6-h transportation group. We also identified several genus-level and amplicon sequence variation-level taxa that displayed significant differences in their abundances across various transportation groups, observed at all sampling time points investigated. This research identifies microbiota changes due to varying transportation durations in surplus dairy calves, providing a broad understanding of the microbial shifts in surplus dairy calves after transportation across varying durations. Although these variations may not directly correlate with overall calf health or indicate dysbiosis, these results emphasize the importance of further investigating transportation practices to enhance calf health and well-being. Further studies are warranted to elucidate the relationship between microbiota and calf health.