Changes In Relative Abundance Research Articles

Conversion of coastal marsh to aquaculture ponds altered soil ammonia oxidiser community and decreased ammonia oxidation potential

AbstractIntroductionAs a crucial component of the nitrogen cycle, ammonia oxidation in soil can be driven by canonical ammonia‐oxidising archaea (AOA) and bacteria, as well as complete ammonia oxidiser (CMX Nitrospira). Land use change can disrupt and alter the soil microbial community and the nitrogen cycle.Materials & MethodsWe compared the soil ammonia‐oxidising microorganisms and ammonia oxidation potentials in a coastal marsh and nearby reclaimed aquaculture ponds, monthly over a 10‐month period in southeastern China. The abundance of ammonia oxidisers was assessed by real‐time quantitative PCR and the community structure of CMX Nitrospira was evaluated by high‐throughput sequencing.ResultsThe ammonia oxidiser community was dominated by AOA in the marsh (91%) and was made up of similar proportions of AOA and CMX Nitrospira in the aquaculture ponds (46%–47%). The CMX Nitrospira community structure changed significantly between habitat types, mainly driven by opposite change in relative abundance of clade B versus clades A.2 and A.3. Aquaculture reclamation decreased the soil potential ammonia oxidation rate (PAO) by an order of magnitude, and AOA was the only significant predictor of PAO among all ammonia oxidiser groups.ConclusionOur results suggest that aquaculture reclamation from coastal marshes would significantly alter the soil ammonia oxidiser community and decrease ammonia oxidation rate, and CMX Nitrospira appear to play a relative larger role in nitrogen cycling in aquaculture ponds.

Cross-Validation of Lipid Structure Assignment Using Orthogonal Ion Activation Modalities on the Same Mass Spectrometer.

The onset and progression of cancer is associated with changes in the composition of the lipidome. Therefore, better understanding of the molecular mechanisms of these disease states requires detailed structural characterization of the individual lipids within the complex cellular milieu. Recently, changes in the unsaturation profile of membrane lipids have been observed in cancer cells and tissues, but assigning the position(s) of carbon-carbon double bonds in fatty acyl chains carried by membrane phospholipids, including the resolution of lipid regioisomers, has proven analytically challenging. Conventional tandem mass spectrometry approaches based on collision-induced dissociation of ionized glycerophospholipids do not yield spectra that are indicative of the location(s) of carbon-carbon double bonds. Ozone-induced dissociation (OzID) and ultraviolet photodissociation (UVPD) have emerged as alternative ion activation modalities wherein diagnostic product ions can enable de novo assignment of position(s) of unsaturation based on predictable fragmentation behaviors. Here, for the first time, OzID and UVPD (193 nm) mass spectra are acquired on the same mass spectrometer to evaluate the relative performance of the two modalities for lipid identification and to interrogate the respective fragmentation pathways under comparable conditions. Based on investigations of lipid standards, fragmentation rules for each technique are expanded to increase confidence in structural assignments and exclude potential false positives. Parallel application of both methods to unsaturated phosphatidylcholines extracted from isogenic colorectal cancer cell lines provides high confidence in the assignment of multiple double bond isomers in these samples and cross-validates relative changes in isomer abundance.

Digestion, nitrogen excretion and rumen microbial communities in beef cattle, and in vitro methane output, of fresh temperate grass with or without citrus pulp supplementation

The effects of supplementing grass with a high-energy, low-protein concentrate on intake, rumen fermentation, rumen microbial communities, apparent total tract digestibility and N-balance in beef cattle, and rumen fermentation and methane output in vitro, were studied. Sixteen Charolais steers (mean live weight (s.d.) 564 (24.3) kg)), were used in a completely randomised block design experiment, and offered either zero-grazed perennial ryegrass (Lolium perenne L.) ad libitum (GO), or the same grass with 2.25 kg citrus pulp dry matter (DM) daily (GCP). During the measurement phase, DM intake was restricted at 0.85 of their pre-determined intake. The same diets were incubated in an eight-vessel RUSITEC system. Supplementation decreased grass DM intake (6.59 vs. 7.54 kg/d) but increased total DM intake by 0.95 kg/d (P < 0.01). Rumen pH (P = 0.05) and NH3 concentration (P < 0.05) decreased, and total volatile fatty acid (VFA) concentrations (P < 0.01), molar proportions of propionate (P = 0.08) and butyrate (P < 0.05) increased for GCP compared to GO. Supplementation significantly altered the overall rumen microbial community structure compared to GO, driven principally by changes in relative abundance of Prevotella, Methoanobrevibacter and Lachnospiraceae genera. Plasma urea concentration was lower (P < 0.05) for GCP than GO. There was no effect (P > 0.10) of supplementation on apparent total tract digestibility of DM, OM and ADFom, but aNDFom digestibility tended to be lower (P = 0.07) and crude protein (N × 6.25) digestibility was lower (P < 0.01) for GCP compared to GO. Nitrogen intake (200 g/d) did not differ (P > 0.05) between treatments. Faeces N loss was 9 g/d greater (P < 0.01), and urine (P < 0.01) and total (P < 0.05) N losses were 22 and 13 g/d lower, respectively, for GCP compared to GO. In terms of N-balance (g/kg N intake), faecal N excretion was greater and urinary N excretion was lower (P < 0.01) resulting in a greater N use efficiency (P = 0.06) for GCP than GO. In vitro rumen NH3 concentrations were lower (P < 0.01) for GCP compared to GO, whereas in vitro rumen pH, lactic acid, VFA concentrations and molar proportions, and methane and total gas output did not differ (P > 0.05) between treatments. Supplementing grass with a low-protein, high-energy feed may be a strategy to reduce urinary and total N excretion in beef cattle grazing temperate grass pasture.

Microbiome variations induced by delta9-tetrahydrocannabinol predict weight reduction in obese mice

IntroductionObesity and high-fat diets induce consistent alterations in gut microbiota composition. Observations from epidemiological reviews and experiments also illustrate weight regulation effects of delta9-tetrahydrocannabinol (THC) with microbiome shifts. Therefore, we investigated the weight-loss potential of THC in obese mice models and to elucidate the contribution of specific gut microbiome changes in THC-induced weight loss.MethodsHigh-fat diet induced obese mice were treated with oral THC supplementation for two weeks and compared with controls. In addition to measuring weight, fecal samples were obtained at various timepoints, sequenced for bacterial 16s rRNA content and analyzed using QIIME2. Alpha and beta diversity were computed followed by linear mixed effects (LME) modeling of bacterial relative abundance relationship to THC treatment and weight change.ResultsIn both male and female mice, the THC group had significantly greater average weight loss than controls (−17.8% vs. −0.22%, p&amp;lt;0.001 and −13.8% vs. +2.9%, p&amp;lt;0.001 respectively). Male mice had 8 bacterial taxonomic features that were both significantly different in relative abundance change over time with THC and correlated with weight change. An LME model using three bacterial features explained 76% of the variance in weight change with 24% of variation explained by fixed effects of feature relative abundance alone. The model also accurately predicted weight change in a second male mouse cohort (R=0.64, R2=0.41, p=&amp;lt;0.001). Female mice had fewer significant predictive features and were difficult to model, but the male-produced 3-feature model still accurately predicted weight change in the females (R=0.66, R2=0.44, p&amp;lt;0.001).ConclusionUsing a stepwise feature selection approach, our results indicate that sex-specific gut microbiome composition changes play some role in THC-induced weight loss. Additionally, we illustrated the concept of microbiome feature-based modeling to predict weight changes.

Relating microbial community composition to treatment performance in an ozone-biologically active carbon filtration potable reuse treatment train

Treatment trains that couple ozone (O3) with biologically active carbon (BAC) filtration are of interest as a lower cost, more sustainable, membrane-free approach to water reuse. However, little is known about the microbial communities that are the fundamental drivers of O3-BAC treatment. The objective of this study was to demonstrate microbial community profiling as a diagnostic tool for assessing the functionality, biological stability, and resilience of coupled physical, chemical, advanced oxidative and biological processes employed in water reuse treatment. We utilized 16S rRNA gene amplicon sequencing to profile the bacterial microbiota over time throughout a potable reuse train employing coagulation, flocculation, sedimentation, ozonation, BAC filtration, granular activated carbon (GAC) adsorption, and UV disinfection. A distinct baseline microbiota was associated with each stage of treatment (ANOSIM, p < 0.05, r-stat = 0.52), each undergoing succession with time and operational shifts. Ozonation resulted in the sharpest shifts (i.e., 83.3 % average change in Genus level relative abundances, when adjusted O3:TOC ratio > 1), and also variance, in microbial community composition. Adjustment in O3:TOC ratios, temperature, filter-aid polymer, monochloramine quenching agent, and empty-bed contact time also resulted in measurable changes in the baseline microbial community composition of individual processes, but to a lesser degree. Of these, supplementation of nitrogen and phosphorus resulted in the strongest bifurcation, especially in the microbial communities inhabiting the BAC (ANOSIM: p < 0.05, BAC5 r-stat = 0.32; BAC10 r-stat = 0.54) and GAC (ANOSIM: p < 0.05, GAC10 r-stat = 0.54; GAC20 r-stat = 0.63) units. Additionally, we found that the BAC microbial community was responsive to an inoculation of microbially active media, which resulted in improved TOC removal. The findings of this study improve understanding of bacterial dynamics occurring in advanced water treatment trains and can inform improved system design and operation.

Linking Aging to Cancer: The Role of Chromatin Biology.

Epigenetic changes have been established to be a hallmark of aging, which implies that aging science requires collaborating with the field of chromatin biology. DNA methylation patterns, changes in relative abundance of histone post-translational modifications, and chromatin remodeling are the central players in modifying chromatin structure. Aging is commonly associated with an overall increase in chromatin instability, loss of homeostasis, and decondensation. However, numerous publications have highlighted that the link between aging and chromatin changes is not nearly as linear as previously expected. This complex interplay of these epigenetic elements during the lifetime of an organism likely contributes to cellular senescence, genomic instability, and disease susceptibility. Yet, the causal links between these phenomena still need to be fully unraveled. In this perspective article, we discuss potential future directions of aging chromatin biology.

Thermonuclear 28P(p, γ)29S reaction rate and astrophysical implication in ONe nova explosion

Context. An accurate 28P(p, γ)29S reaction rate is crucial to defining the nucleosynthesis products of explosive hydrogen burning in ONe novae. Using the recently released nuclear mass of 29S, together with a shell model and a direct capture calculation, we reanalyzed the 28P(p, γ)29S thermonuclear reaction rate and its astrophysical implication. Aims. We focus on improving the astrophysical rate for 28P(p, γ)29S based on the newest nuclear mass data. Our goal is to explore the impact of the new rate and associated uncertainties on the nova nucleosynthesis. Methods. We evaluated this reaction rate via the sum of the isolated resonance contribution instead of the previously used Hauser-Feshbach statistical model. The corresponding rate uncertainty at different energies was derived using a Monte Carlo method. Nova nucleosynthesis is computed with the 1D hydrodynamic code SHIVA. Results. The contribution from the capture on the first excited state at 105.64 keV in 28P is taken into account for the first time. We find that the capture rate on the first excited state in28 P is up to more than 12 times larger than the ground-state capture rate in the temperature region of 2.5 × 107 K to 4 × 108 K, resulting in the total 28P(p, γ)29S reaction rate being enhanced by a factor of up to 1.4 at ~1 × 109 K. In addition, the rate uncertainty has been quantified for the first time. It is found that the new rate is smaller than the previous statistical model rates, but it still agrees with them within uncertainties for nova temperatures. The statistical model appears to be roughly valid for the rate estimation of this reaction in the nova nucleosynthesis scenario. Using the 1D hydrodynamic code SHIVA, we performed the nucleosynthesis calculations in a nova explosion to investigate the impact of the new rates of 28P(p, γ)29S. Our calculations show that the nova abundance pattern is only marginally affected if we use our new rates with respect to the same simulations but statistical model rates. Finally, the isotopes whose abundance is most influenced by the present 28P(p, γ)29S uncertainty are 28Si, 33,34S, 35,37Cl, and 36Ar, with relative abundance changes at the level of only 3% to 4%.

Comparative analysis of the structural and compositional change of spotted sea bass (Lateolabrax maculatus) gut microflora following Aeromonas veronii infection and the effects of Lactobacillus plantarum on these changes

Growing evidence suggests a close relationship between gut microbiota and infectious diseases. However, the specific role of gut microbiota in host-pathogen interactions during aquaculture-related infections remains poorly understood. This study investigated the diversity and composition of gut microbiota communities in Aeromonas veronii-infected Lateolabrax maculatus using high-throughput sequencing. The results revealed significant changes in the structure and composition of L. maculatus gut microbiota after A. veronii infection. Over time, Bacteroidetes and Firmicutes decreased significantly, while Proteobacteria increased significantly after A. veronii infection. Most intestinal bacteria showed a decline in abundance over time, with probiotics (such as Lactobacillus) experiencing a significant decrease and pathogens (such as Aeromonas) showing a significant increase. Conversely, no differences were observed in the structure and composition of gut microbiota between healthy L. maculatus and those infected with A. veronii after treatment with Lactobacillus plantarum; no changes in relative abundances of other bacterial phyla or genera except for Aeromonas. Furthermore, intestinal flora’s structural diversity and composition differed significantly from untreated L. maculatus infected with A. veronii. These findings suggest alterations in the structure and composition of gut microbiota following A. veronii infection. L. plantarum can maintain a dynamic balance within the intestinal flora, reducing the potential risk of pathogen infections.

Detecting mining impacts on freshwater ecosystems using replicated sampling before and after the impact

Detecting human impact on freshwater ecosystems is problematic without rigorous assessment of temporal changes. Assessments of mining impacts are further complicated by the strong influence of local catchment geology on surface waters even in unmined environments. Such influence cannot be effectively considered by using broad-scale reference frameworks based on regionalization and stream types. Using the BACI (Before-After Control-Impact) design, we examined the impact of mining discharges on freshwater algae and macroinvertebrate communities resulting from the rerouting of treated wastewaters through a pipeline to larger water bodies in Northern and North-Eastern Finland. Impacted sites and control sites were sampled 1 to 2 years before and 1 to 3 years after the pipelines became operational. Stream diatom communities recovered from past loadings upstream of the pipeline (which was no longer impacted by wastewaters) after rerouting of the wastewaters, while no changes downstream from the pipeline were detected. Upstream from the pipeline, diatom species richness increased and changes in relative abundances of the most common diatom taxa as well as in the overall community composition were observed. The effects of the pipeline were less evident for stream macroinvertebrate communities. There was an indication that regional reference conditions used in national biomonitoring may not represent diatom communities in areas with a strong geochemical background influence. Lake profundal macroinvertebrate communities were impacted by past loadings before the construction of the pipeline, and the influence of the pipeline was observed only as changes in the abundances of a few individual species such as phantom midges (which increased in abundance in response to discharges directed through the pipeline). Our results highlight the variable influence of mining discharges on aquatic communities. Statistically strong monitoring programmes, such as BACI designs, are clearly needed to detect these influences.

Chemotherapy-induced gut microbiome disruption, inflammation, and cognitive decline in female patients with breast cancer

Chemotherapy is notorious for causing behavioral side effects (e.g., cognitive decline). Notably, the gut microbiome has recently been reported to communicate with the brain to affect behavior, including cognition. Thus, the aim of this clinical longitudinal observational study was to determine whether chemotherapy-induced disruption of the gut microbial community structure relates to cognitive decline and circulating inflammatory signals. Fecal samples, blood, and cognitive measures were collected from 77 patients with breast cancer before, during, and after chemotherapy. Chemotherapy altered the gut microbiome community structure and increased circulating TNF-α. Both the chemotherapy-induced changes in microbial relative abundance and decreased microbial diversity were related to elevated circulating pro-inflammatory cytokines TNF-α and IL-6. Participants reported subjective cognitive decline during chemotherapy, which was not related to changes in the gut microbiome or inflammatory markers. In contrast, a decrease in overall objective cognition was related to a decrease in microbial diversity, independent of circulating cytokines. Stratification of subjects, via a reliable change index based on 4 objective cognitive tests, identified objective cognitive decline in 35% of the subjects. Based on a differential microbial abundance analysis, those characterized by cognitive decline had unique taxonomic shifts (Faecalibacterium, Bacteroides, Fusicatenibacter, Erysipelotrichaceae UCG-003, and Subdoligranulum) over chemotherapy treatment compared to those without cognitive decline. Taken together, gut microbiome change was associated with cognitive decline during chemotherapy, independent of chemotherapy-induced inflammation. These results suggest that microbiome-related strategies may be useful for predicting and preventing behavioral side effects of chemotherapy.

Yarrowia lipolytica yeast as a dietary supplement for rainbow trout (Oncorhynchus mykiss): Effects on gut microbiota, health and immunity

This study evaluated the effect of whole and autolysed Yarrowia lipolytica CBS 7504 strain on growth performance, gut immunity and gut microbiota composition in juvenile rainbow trout (Oncorhynchus mykiss). Fish were fed diets containing 2% and 5% whole or autolysed Y. lipolytica (WY and AY) in a 45-day trial, with a no-yeast diet as control. Log2 fold change analysis of Y. lipolytica diets showed relative change in abundance and association of certain microbial taxa with respect to control such as a correlation between the relative abundance of Desulfovibrionaceae in diets with WY. Moreover, diets at 5% and 2% levels demonstrated a synergistic association with relative levels of Sphingobacteriaceae and Rhodobacteraceae, respectively. However, the overall analyses showed little/no impact of Y. lipolytica supplementation on overall intestinal microbial composition, growth parameters and body indices. Gene expression analysis of the intestine revealed significant elevation in expression of immune-related genes of the complement pathway (c3 and c-type lectin), membrane receptor pathway (tlr2 and tlr5), mucosal innate immune pathway (muc-2), cytokines (tnf-α) and adaptive immune pathway (igt and cd4) in 5% WY. In conclusion, it can be inferred that dietary Y. lipolytica can be a useful functional feed supplement for rainbow trout when used as whole yeast at a 5% inclusion level since it can modulate gut microbial communities and act as a potential immunostimulant for the host.

ARTseq-FISH reveals position-dependent differences in gene expression of micropatterned mESCs

Differences in gene-expression profiles between individual cells can give rise to distinct cell fate decisions. Yet how localisation on a micropattern impacts initial changes in mRNA, protein, and phosphoprotein abundance remains unclear. To identify the effect of cellular position on gene expression, we developed a scalable antibody and mRNA targeting sequential fluorescence in situ hybridisation (ARTseq-FISH) method capable of simultaneously profiling mRNAs, proteins, and phosphoproteins in single cells. We studied 67 (phospho-)protein and mRNA targets in individual mouse embryonic stem cells (mESCs) cultured on circular micropatterns. ARTseq-FISH reveals relative changes in both abundance and localisation of mRNAs and (phospho-)proteins during the first 48 hours of exit from pluripotency. We confirm these changes by conventional immunofluorescence and time-lapse microscopy. Chemical labelling, immunofluorescence, and single-cell time-lapse microscopy further show that cells closer to the edge of the micropattern exhibit increased proliferation compared to cells at the centre. Together these data suggest that while gene expression is still highly heterogeneous position-dependent differences in mRNA and protein levels emerge as early as 12 hours after LIF withdrawal.

Prospective manipulation of the gut microbiome with microbial ecosystem therapeutic 4 (MET4) in HPV-related locoregionally-advanced oropharyngeal cancer squamous cell carcinoma (LA-OPSCC) undergoing primary chemoradiation: ROMA2 study.

Gut microbiome modulation to boost antitumor immune responses is under investigation. ROMA-2 evaluated the microbial ecosystem therapeutic (MET)-4 oral consortia, a mixture of cultured human stool-derived immune-responsiveness associated bacteria, given with chemoradiation (CRT) in HPV-related oropharyngeal cancer patients. Co-primary endpoints were safety and changes in stool cumulative MET-4 taxa relative abundance (RA) by 16SRNA sequencing. Stools and plasma were collected pre/post-MET-4 intervention for microbiome and metabolome analysis. Twenty-nine patients received ≥1 dose of MET-4 and were evaluable for safety: drug-related adverse events (AEs) occurred in 13/29 patients: all grade 1-2 except one grade 3 (diarrhea). MET-4 was discontinued early in 7/29 patients due to CRT-induced toxicity, and in 1/29 due to MET-4 AEs. Twenty patients were evaluable for ecological endpoints: there was no increase in stool MET-4 RA post-intervention but trended to increase in stage III patients (p = 0.06). MET-4 RA was higher in stage III vs I-II patients at week 4 (p = 0.03) and 2-month follow-up (p = 0.01), which correlated with changes in plasma and stool targeted metabolomics. ROMA-2 did not meet its primary ecologic endpoint, as no engraftment was observed in the overall cohort. Exploratory findings of engraftment in stage III patients warrants further investigation of microbiome interventions in this subgroup.

Exertional heat stress promotes the presence of bacterial DNA in plasma: A counterbalanced randomised controlled trial

ObjectivesThe primary aim was to explore the impact of exertional-heat stress (EHS) promoted exercise-associated bacteraemia. A secondary aim was to examine if an amino acid beverage (AAB) intervention may mitigate exercise-associated bacteraemia. DesignCounterbalanced randomised control trial. MethodsTwenty endurance trained male participants completed two randomised EHS trials. On one occasion, participants consumed a 237 mL AAB twice daily for 7 days prior, immediately before and every 20 min during EHS (2 h running at 60 % V̇O2max in 35 °C). On the other occasion, a water volume control (CON) equivalent was consumed. Whole blood samples were collected pre- and immediately post-EHS, and were analysed for plasma DNA concentration by fluorometer quantification after microbial extraction, and bacterial relative abundance by next generation 16s rRNA gene sequencing. ResultsIncreased concentration of microbial DNA in plasma pre- to post-EHS was observed on CON (pre-EHS 0.014 ng/μL, post-EHS 0.039 ng/μL) (p < 0.001) and AAB (pre-EHS 0.015 ng/μL, post-EHS 0.031 ng/μL) (p < 0.001). The magnitude of change from pre- to post-exercise on AAB was 40 % lower, but no significant difference was observed versus CON (p = 0.455). Predominant bacterial groups identified included: phyla-Proteobacteria (88.0 %), family-Burkholderiaceae (59.1 %), and genus-Curvibacter (58.6 %). No significant variation in absolute and relative change in α-diversity and relative abundance for phyla, family, and genus bacterial groups was observed in AAB versus CON. ConclusionsThe increased presence of microbial-bacterial DNA in systemic circulation in response to EHS appears positive in all participants. An amino acid beverage supplementation period prior to and consumption during EHS did not provide significant attenuation of EHS-associated bacteraemia.

Seasonal changes of ammonia-oxidizing bacterial communities during tropical forest restoration

In this study, we used a high-throughput sequencing technology to survey the dry-wet seasonal change characteristics of soil ammonia-oxidizing bacteria (AOB) communities in the three restoration stages [i.e., Mallotus paniculatus community (early stage), Millettia leptobotrya community (middle stage), and Syzygium oblatum community (later stage)] of Xishuangbanna tropical forest ecosystems. We analyzed the effects of soil physicochemical characteristics on AOB community composition and diversity during tropical forest restoration. The results showed that tropical forest restoration significantly affected the relative abundance of dominant AOB phyla and their dry-wet seasonal variation. The maximum relative abundance of Proteobacteria (71.3%) was found in the early recovery stage, while that of Actinobacteria was found in the late recovery stage (1.0%). The abundances of Proteobacteria and Actinobacteria had the maximum ranges of dry-wet seasonal variation in the early and late stages, respectively. The abundance of dominant AOB genera and its dry-wet seasonal variation varied across tropical forest restoration stages. The maximum average relative abundance of Nitrosospira and Nitrosomonas in the late recovery stage was 66.2% and 1.5%, respectively. In contrast, the abundance of Nitrosovibrio reached its maximum (25.6%) in the early recovery stage. The maximum dry-wet seasonal variation in relative abundance of Nitrosospira and Nitrosomonas occurred in the early recovery stage, while that of Nitrosovibrio occurred in the middle recovery stage. The Chao1, Shannon, and Simpson diversity indices of AOB communities increased along the restoration stages, which were significantly higher in the wet season than in the dry season. The results of canonical correspondence analysis showed that soil easily oxidized carbon was the main factor controlling AOB community diversity and Actinobacteria abundance. Soil bulk density and temperature were the main factors affecting Proteobacteria abundance. Soil pH, microbial biomass carbon, water content, ammonium nitrogen, bulk density, and temperature were the main factors controlling the abundances of Nitrosospira, Nitrosomonas, and Nitrosovibrio. Therefore, tropical forest restoration can regulate the change of relative abundance of dominant AOB taxa via mediating the changes of soil temperature, bulk density, and readily oxidized carbon, leading to an increase in soil AOB community diversity.

Host influence on the eukaryotic virome of sympatric mosquitoes and abundance of diverse viruses with a broad host range.

Mosquitoes harbor a large diversity of eukaryotic viruses. Those viromes probably influence mosquito physiology and the transmission of human pathogens. Nevertheless, their ecology remains largely unstudied. Here, we address two key questions in virome ecology. First, we assessed the influence of mosquito species on virome taxonomic diversity and relative abundance. Contrary to most previous studies, the potential effect of the habitat was explicitly included. Thousands of individuals of Culex poicilipes and Culex tritaeniorhynchus, two vectors of viral diseases, were concomitantly sampled in three habitats over two years. A total of 95 viral taxa from 25 families were identified with meta-transcriptomics, with 75% of taxa shared by both mosquitoes. Viromes significantly differed by mosquito species but not by habitat. Differences were largely due to changes in relative abundance of shared taxa. Then, we studied the diversity of viruses with a broad host range. We searched for viral taxa shared by the two Culex species and Aedes vexans, another disease vector, present in one of the habitats. Twenty-six out of the 163 viral taxa were found in the three mosquitoes. These taxa encompassed 14 families. A database analysis supported broad host ranges for many of those viruses, as well as a widespread geographical distribution. Thus, the viromes of mosquitoes from the same genera mainly differed in the relative abundance of shared taxa, whereas differences in viral diversity dominated between mosquito genera. Whether this new model of virome diversity and structure applies to other mosquito communities remains to be determined.

The endophytic microbiome response patterns of Juglans regia to two pathogenic fungi.

The endophytic microbial community reassembles to participate in plant immune balance when the host plants are stressed by pathogens. However, it remains unclear whether this assembly is pathogen-specific and how regulatory pathways are coordinated in multi-pathogens. In order to investigate the effects of infection with Colletotrichum gloeosporioides (Cg treatment) and Fusarium proliferatum (Fp treatment) on walnut leaf endophytic microbiome in their assembly, co-occurrence pattern, and on comprehensive chemical function of the internal environment of leaf, an interaction system of the walnut-pathogenic fungi was constructed using seed embryo tissue culture technology. The study showed differences in the assembly of endophytic microbial communities in walnut trees across three groups (control group, Ck; Cg; Fp) after Cg and Fp treatments. Despite changes in relative abundances, the dominant communities in phyla and genera remained comparable during the infection of the two pathogens. Endophyte fungi were more sensitive to the pathogen challenge than endophyte bacteria. Both promoted the enrichment of beneficial bacteria such as Bacillus and Pseudomonas, changed the modularity of the community, and reduced the stability and complexity of the endophyte community. Pathogenic fungi infection mainly affects the metabolism of porphyrin and chlorophyll, purine metabolism, phenylpropane metabolism, and amino acid metabolism. However, there was no significant difference in the secondary metabolites for the different susceptible plants. By screening endogenous antagonistic bacteria, we further verified that Pseudomonas psychrotolerans and Bacillus subtilis had inhibitory effects on the two pathogenic fungi and participated in the interaction between the leaves and pathogenic fungi. The antibacterial substances may be 1-methylnaphthalene, 1,3-butadiene, 2,3-butanediol, and toluene aldehyde.

Proteomic biomarkers in seminal plasma as predictors of reproductive potential in azoospermic men.

Azoospermia, characterized by an absence of sperm in the ejaculate, represents the most severe form of male infertility. While surgical sperm retrieval in obstructive azoospermia (OA) is successful in the majority of cases, patients with non-obstructive azoospermia (NOA) show retrieval rates of only about 50% and thus frequently have unnecessary surgery. Surgical intervention could be avoided if patients without preserved spermatogenesis are identified preoperatively. This prospective study aimed to discover biomarkers in seminal plasma that could be employed for a non-invasive differential diagnosis of OA/NOA in order to rationalize surgery recommendations and improve success rates. All patients signed written informed consent, underwent comprehensive andrological evaluation, received human genetics to exclude relevant pathologies, and patients with azoospermia underwent surgical sperm retrieval. Using label-free LC-MS/MS, we compared the proteomes of seminal plasma samples from fertile men (healthy controls (HC), n=8) and infertile men diagnosed with 1) OA (n=7), 2) NOA with successful sperm retrieval (mixed testicular atrophy (MTA), n=8), and 3) NOA without sperm retrieval (Sertoli cell-only phenotype (SCO), n=7). Relative abundance changes of two candidate markers of sperm retrieval, HSPA2 and LDHC, were confirmed by Western Blot. We found the protein expression levels of 42 proteins to be significantly down-regulated (p ≤ 0.05) in seminal plasma from SCO NOA patients relative to HC whereas only one protein was down-regulated in seminal plasma from MTA patients. Analysis of tissue and cell expression suggested that the testis-specific proteins LDHC, PGK2, DPEP3, and germ-cell enriched heat-shock proteins HSPA2 and HSPA4L are promising biomarkers of spermatogenic function. Western blotting revealed a significantly lower abundance of LDHC and HSPA2 in the seminal plasma of men with NOA (SCO and MTA) compared to controls. The results indicate that certain testis-specific proteins when measured in seminal plasma, could serve as indicators of the presence of sperm in the testis and predict the success of sperm retrieval. Used in conjunction with conventional clinical assessments, these proteomic biomarkers may assist in the non-invasive diagnosis of idiopathic male infertility.

Environmental DNA based biomonitoring for hatchery-raised fish in riverine habitats before and after recordable flood event

It is reported that the magnitudes of flood events in riverine systems have been increasing due to global climate change. Because flood events could displace fish species downstream and/or increase the mortality of fish, it is important to know how the increased floods can affect fish in river networks. In this study, we focused a hatchery-raised amphidromous fish, Ayu Plecoglossus altivelis altivelis in a mainstem-tributary network in Japan and examined the relative fish abundance changes before and after the recordable massive flood using the environmental DNA (eDNA) analysis. We also examined the spatiotemporal patterns of the Ayu eDNA concentrations and the relationships with tributary discharge, width, and depth to examine if the relative abundance changes could be related to tributary size. Our results indicated that Ayu tended to inhabit large, deep tributaries more than small, shallow tributaries even after the flood event. Further, the eDNA concentrations of Ayu have decreased in most study sites after the flood; however, the eDNA concentrations in certain tributaries with lower tributary size have increased during the study period. These results suggest that (1) the habitat conditions could be important for the maintenance of Ayu populations before and after flood events, (2) increase in flood magnitude along climate changes could have impacts on Ayu populations, and (3) not only large tributary, but also small tributaries could be important habitats for the target species to avoid flood disturbances.

460 Changes in Lipid Profiles with Progression of Pregnancy in Black Women

OBJECTIVES/GOALS: Pregnant African American (Black women) have higher rates of adverse pregnancy outcomes compared to other races and routine monitoring of lipid levels is not currently in practice in prenatal care. We hypothesized that lipid profiles would show variation across pregnancy indicative of specific requirements during gestation and fetal development. METHODS/STUDY POPULATION: We used an untargeted lipidome analysis approach to investigate lipid metabolism with the progression of pregnancy. Pregnant Black women were recruited at prenatal clinics in Midwest (Metro Detroit, Michigan and Columbus, Ohio), women under 18 or above 45 years of age were not enrolled due to metabolic changes associated with these age groups. Women signed the consent forms and plasma samples were collected at 8-18 weeks at (T1), 22-29 weeks (T2) and 30-36 weeks (T3) of pregnancy. Samples from sixty-three women (mean age 27.41 ± 5.61 years) who had term birth (completed 37 weeks of pregnancy) were subjected to “shotgun” Orbitrap high resolution/ accurate mass spectrometry. Mixed-effects models were used to quantify systematic changes in relative lipid abundances over time using R lme4 and ggplot2 packages. RESULTS/ANTICIPATED RESULTS: Total lipids and some major lipid classes showed a significant increase with the progression of pregnancy. Phospholipids and glycerolipids exhibited a gradual increase throughout pregnancy, while sphingolipids and total sterol lipids displayed a more pronounced increase at the T3 timepoint. Acylcarnitines, hydroxy acylcarnitines and Lyso phospholipids levels significantly decrease from T1 to T3. One of the interesting finding was that non-esterified fatty acids decreased from T1 to T2 and increased again from T2 to T3, suggesting a possible role for these lipids during the later stages of pregnancy. The fatty acids showing this trend included key fatty acids- Linoleic Acid, Arachidonic Acid, Alpha-linolenic acid, Eicosapentaenoic acid, Docosapentaenoic acid, Docosahexaenoic acid. DISCUSSION/SIGNIFICANCE: Mapping lipid trends during pregnancy could lend support to a precision health approach to reduce perinatal health disparities among pregnant Black women. The findings from this study will be used to identify biomarkers and study associations with social and environmental factors responsible for adverse perinatal outcome in pregnant Black women.