Fecal Samples Research Articles

Exposure to greenspaces sourced soils improves mice gut microbiota

Greenspaces are important components of our living environment and have been linked to various human health. However, the mechanisms underlying the linkages remain unclear. Enriching microbiota has emerged as a novel mechanism, but the corresponding evidence is still limited. We collected soil samples from forest land, grassland, and barren land in Zunyi City, southwestern China and prepared soil solutions. A total of 40 BALB/c mice were evenly divided into normal control group, model control group, forest soil group, grassland soil group, and barren land soil group. After establishing the pseudo germ-free mouse model, different soil solutions were administered through gavage, lasting for seven weeks. Fecal samples were collected and a 16S rRNA high-throughput sequencing analysis was performed. Then, alpha- and beta-diversity were calculated and employed to estimate the effects of soil exposures on mice gut microbial diversity and composition. Further, Linear Discriminant Analysis Effect Size (LEfSe) analysis was carried out to evaluate the effects of soil exposures on gut microbiota specific genera abundances and functional pathways. Compared to mice exposed to barren land soils, those exposed to soils sourced from forest land showed an increase of 0.43 and 70.63 units in the Shannon index and the Observed ASVs, respectively. In addition, exposure to soils sourced from forest land and grassland resulted in healthier changes (i.e., more short-chain fatty acids (SCFAs)-producing bacteria) in gut microbiota than those from barren land. Furthermore, mice exposed to forest soil and grassland soil showed enrichment in 5 and 3 pathways (e.g., butanoate metabolism) compared to those exposed to barren land soil, respectively. In conclusion, exposure to various greenspaces soils may modify the gut microbial communities of mice, potentially fostering a more beneficial microbiota profile. Further research is needed to explore the metabolites of these gut microbiota changes and their correlations with human health.

Supplementation of lysophospholipids in calf starter: effects on growth performance, blood metabolites, health, and ruminal ammonia nitrogen concentration

The aim of the current study was to evaluate the effects of a commercial mixture of Lysophospholipids (LPL) in calf starter on performance, blood metabolites, antioxidant capacity, liver enzymes concentration, and health status of pre-weaning dairy calves. Forty-eight newborn female Holstein calves (37.5 ± 2.9kg of body weight) were randomly assigned to four treatments (12 replicates), which included the control (CON, without additive) and a commercial blend of LPL at 0.5, 1, and 1.5g/kg of starter feed. The body weight of the calves was measured using a digital scale on days 15, 30, 45, and d 63 of the experimental period to calculate the average daily gain (ADG). The individual starter intake and fecal score were measured daily. Blood samples were collected at 30 and 63 days, while fecal samples were collected for four consecutive days after weaning to measure nutrient digestibility. The treatments did not significantly afffect the total dry matter and starter intake. However, the supplementation of LPL resulted in an increase in the ADG and improved feed efficiency (FE, from1-63 d) compared to the CON group. The addition of LPL did not have any significant impact on the apparent digestibility of dry matter (DM), and organic matter (OM). Eether extract (EE) digestibility tended to increase with the addition of LPL. Calves that received starter feed containing 0.5g/kg of LPL had higher superoxide dismutase activity at 30 d of age compared to the intake.CON group. Additionally, the average fecal score significantly decreased in calves fed 0.5 and 1.5g/kg LPL compared with the CON group. These findings suggest that the inclusion of LPL at an average dose of 1g/kg of calf starter can improve the performance of pre-weaning Holstein dairy calves.

Giardia duodenalis in sympatric wild reindeer and domestic sheep in Norway

Wild and semi-domesticated reindeer graze freely on natural pastures in Norway, often sharing these with domestic sheep and other domestic and wild ruminants. In this study, faecal samples from wild reindeer and domestic sheep were collected from two areas in southern Norway and analysed to assess the occurrence and assemblage of Giardia duodenalis. Among 162 wild reindeer samples, 25 (15%) were positive for Giardia, showing high infection intensities, with most of the samples belonging to sub-assemblage AI, which has zoonotic potential. Interestingly, this study did not detect subassemblage AIII, known to be found in wild ruminants. Among 45 sheep samples, 13 (29%) were Giardia-positive, with most belonging to assemblage E.The finding of predominantly assemblage AI in the reindeer was surprising, particularly given the large proportion of sheep shedding assemblage E Giardia cysts. As the number of sheep on these natural pastures far outnumbers the wild reindeer, it is intriguing that they do not seem to share Giardia assemblages.

Lactiplantibacillus plantarum P101-fermented Luffa cylindrica Juice Alleviate F-53B-Induced Liver Steatosis in Mice via the Gut-liver axis

6:2 chlorinated polyfluorinated ether sulfonate (F-53B) is a novel food contaminant that can accumulate in organisms within the food chain, posing significant risks to human health. Currently, there are no safe and effective measures to alleviate the injury caused by F-53B, particularly liver injury. Based on the health-promoting potential of fermented foods, this study aims to assess whether Lactiplantibacillus plantarum P101-fermented Luffa cylindrica juice (LLJ) exerts alleviating effects against F-53B-induced liver injury in mice. Following orally administering 10 mL/kg body weight/day of LLJ to C57BL/6J male mice exposed to F-53B at 1 mg/kg body weight/day for 56 days, liver, colon, blood, and fecal samples were collected to assess liver damage, gut function and microbiota composition, and short-chain fatty acids (SCFAs) levels. LLJ treatment effectively alleviated F-53B-induced liver function decline and liver steatosis. Moreover, LLJ treatment effectively alleviated F-53B-induced colon injury and gut microbiota dysbiosis. Crucially, fecal microbiota transplantation experiment demonstrated the crucial role of LLJ-modulated gut microbiota in alleviating F-53B-induced liver injury. Further analysis indicated that the alleviating effects of LLJ may be mediated through increased SCFA production by the gut microbiota. These findings suggest that adding fermented products to the diet may alleviate the adverse effects of emerging food contaminants. Our work offers new perspectives on the potential of fermented products as dietary interventions to counteract the adverse effects of contaminants like F-53B, offering promising avenues for dietary therapy.

Effects of Lambda-cyhalothrin on Gut Microbiota and Related Bile Acid Metabolism in Mice

Since the gut microbiota plays a crucial role in host metabolism and homeostasis, its alterations induced by xenobiotics such as pesticides, could pose a risk to host health. The pyrethroid insecticides were frequently detected in surface water (up to 13mg/L worldwide), sediments, and agricultural products; additionally, some previous studies indicated that pyrethroid insecticides could cause disruption of gut homeostasis. Hence herein, the normally used pyrethroid lambda-cyhalothrin (LCT) was selected and studied for its effects on the intestinal microbial community and its related bile acid metabolism using mice as the model species. Results showed that the total amount of bile acids in plasma and fecal samples from LCT treated mice markedly increased compared to controls, which could be mainly ascribed to the significantly raised proportions of taurine conjugated bile acids in plasma, and the increase in fecal secondary bile acids. In gut microbial profiles, a significantly increased richness of Prevotellacea and a depletion of Lachnospiraceae were found at the family level upon the treatment with lambda-cyhalothrin. In conclusion, results obtained on bacterial and bile acid profiles corroborate that the treatment of mice with LCT could affect gut microbial community with accompanying changes in bile acid homeostasis. Environmental ImplicationPesticides, especially the widely used pyrethroids have been extensively reported on their toxicity and the accumulation of pyrethroids in environment could pose a risk to animal and human health. Our study is the first to report the impact of one normally used pyrethroid lambda-cyhalothrin on gut microbiota and it mediated bile acid metabolism. Results showed that lambda-cyhalothrin could significantly alter the fecal bacterial and bile acid profiles even at a low dose, suggesting the reduction in amount and frequency for the use of pyrethroids, which not only benefits to public health but also protects our living surroundings.

Dairy farm waste: a potential reservoir of diverse antibiotic resistance and virulence genes in aminoglycoside- and beta-lactam-resistant Escherichia coli in Gansu Province, China

Aminoglycosides (AGs) and beta-lactams are the most commonly used antimicrobials in animal settings, particularly on dairy farms. Dairy farm waste is an important reservoir of antibiotic resistance genes (ARGs) and virulence genes (VGs) in environmental Escherichia coli, which is an important indicator of environmental contamination and foodborne pathogen that potentially threaten human and animal health. In the present study, we aimed to characterize the ARGs and VGs in AG- and beta-lactam-resistant E. coli from dairy farm waste in Gansu Province, China. The dairy farm waste consisted of fecal (n=265) and sewage (n=54) samples processed using standard microbiological techniques and the Clinical & Laboratory Standards Institute guidelines. The total DNA of AG- and beta-lactam-resistant E. coli was extracted, and whole-genome sequencing (WGS) was performed using the Illumina NovaSeq platform and analyzed using various bioinformatics tools. In this study, among 84.3% (269/319) of the E. coli strains, 23.8% (64/269) were identified as AG- and beta-lactam-resistant E. coli. WGS analysis revealed a large pool of ARGs belonging to multiple classes such as AGs, beta-lactams, aminocoumarins, fluoroquinolones, macrolides, phenicol, tetracyclines, phosphonic acid, disinfecting and antiseptic agents, elfamycin, rifamycin, and multidrug resistance genes. Furthermore, virulome analysis of 64 E. coli strains revealed clinically important virulence factors associated with adherence, biofilm, invasion, auto-transportation, siderophores, secretion systems, toxins, anti-phagocytosis, quorum sensing, regulation, metabolism, and motility. We identified dairy farm feces and sewage waste as important reservoirs of antimicrobial resistance and virulence determinants in E. coli in Gansu, China, which can threaten human and animal health through ecological exposure and ingestion of contaminated food and water. We recommend continuous large-scale surveillance in dairy farm settings to formulate protective guidelines for public health safety.

Mulberry leaf extract ameliorates high-fat diet-induced obesity in mice by regulating the gut microbiota and metabolites

The increasing prevalence of obesity presents a notable challenge to global health. Mulberry leaves enriched with biologically active substances hold promise for alleviating obesity. This research assessed the efficacy of mulberry leaf extract (MLE) in mitigating obesity and metabolic disturbances in mice fed a high-fat diet (HFD). The results demonstrated that MLE, administered at both low (167 mg/kg) and high (333 mg/kg) doses, decreased weight gain and fat accumulation, improved glucose metabolism, enhanced insulin sensitivity, and reduced inflammation. Analysis of fecal samples via 16S rRNA sequencing revealed that MLE corrected the gut microbiota imbalance caused by HFD, lowering the Firmicutes/Bacteroidota ratio as well as increasing the relative abundances of Bifidobacterium, Faecalibaculum, Coriobacteriaceae_UCG-002, and Dubosiella. Additionally, serum non-targeted metabolomics analysis identified 19 key differential metabolites associated with MLE treatment, which were primarily related to glycerophospholipid metabolism and tryptophan metabolism. Correlation analysis revealed negative associations between Dubosiella and Coriobacteriaceae_UCG-002 with specific phospholipids, and positive associations of 5-hydroxyindoleacetate and quinoline-4,8-diol with Bifidobacterium and Coriobacteriaceae_UCG-002. These findings suggest that MLE is effective in treating obesity and underscores its potential as a functional food.

Molecular patterns of microbial and metabolic interactions in septic patients with persistent lymphopenia

BackgroundPersistent lymphopenia can be regarded as an important index of acquired immune dysfunction in sepsis. Whether the specific immune factor changes in septic patients with lymphopenia and the correlation to gut microbiota and metabolites remain unclear. MethodsThis single-center prospective observation conducted lymphocyte subgroup analysis of blood samples and 16S rRNA gene amplicons sequencing and untargeted metabolomics analysis of fecal samples from 36 subjects with the persistent (≥3d) (n=21) and non-persistent lymphopenia (<3d) (n=15). ResultsThe persistent lymphopenia showed higher the 28d mortality and 90d mortality, while significantly lower CD3+T/LY, CD3+T cells, CD3+CD4+T cells, CD3+CD8+T cells, Th1 cells, Th2 cells, CD45RA+Treg cells. The 16S rRNA results showed that Staphylococcus, Peptostreptococcus, Bulleidia, Leuconostoc were significant enriched in the persistent lymphopenia. The metabolomics analysis showed that α-Ketoisovaleric acid was increased and 7-DHCA, α-MCA, β-MCA, HCA, LCA-3S, CA, UCA and Citramalic acid were decreased in the persistent lymphopenia. ConclusionIn the process of interaction between host receptors and gut microbiota in patients with persistent lymphopenia sepsis, with a significant reduction in gut microbiota diversity and bile acid metabolites. That can affect various inflammatory pathways of gut immune cells, causing immune dysfunction in the body, which may be one of the main causes of death.

Dietary analysis using prey-DNA metabarcoding from fecal samples of the generalist arthropod predator, Eremias argus

Species-specific dietary information contributes to the conservation of biodiversity and an understanding of the trophic structure of habitats. To investigate the diet of the Mongolian racerunner (Eremias argus), classified as an endangered species in the Republic of Korea, we analyzed its fecal samples using metabarcoding with the arthropod mitochondrial cytochrome c oxidase subunit I (COI) universal primer. In our study, 53 prey taxa were detected in 41 genera, 26 families, 8 orders, and 2 classes (Insecta and Arachnida), and 30 (56.6 %) were identified at the species level. Lepidoptera were the most abundant prey source, accounting for 57.4 % of the total identified prey taxa, followed by Coleoptera (11.3 %), Orthoptera (7.5 %), and Araneae (7.5 %). Compared to previous studies, our approaches were more efficient in detecting various and specific prey types of E. argus, known as a generalist predator. We also discussed the shortcomings of metabarcoding dietary analysis in brief. Overall, our results show that the fecal DNA metabarcoding approach is appropriate for studying the diet of E. argus, an arthropod generalist, and that our results may contribute to the conservation of this endangered species.

Parasitic intestinal infection in students of disabilities centers in Sanaa governorate, Yemen

Background: Intestinal parasite infections remain prevalent in low-income countries, especially among children, due to poor drinking water quality and insufficient personal and environmental cleanliness. This study sought to diagnose, prevent, and manage intestinal parasites in individuals with special needs in Sanaa Governorate. Method: We collected stool samples from 232 students, aged 1 to 18, of which 143 were male and 89 were female. We collected data using a validated standard questionnaire. We processed and analyzed fecal samples using a wet mount preparation, adhering to the standing concentration technique. Results: We found an overall prevalence of intestinal parasitic infection of 83.6%. E. histolytica (76.7%), G. lamblia (8.6%), A. lumbricoides (0.4%), H. nana (6.5%), S. mansoni (0.4%), Ent. vermicularis (6.5%), and S. stercoralis (2.2%). Conclusions: In children, E. histolytica was the most common intestinal parasite infection. There are few methods of transmission and protection for students with special needs.

Feeding ecology of two wild sympatric canids in protected areas of northeastern Argentina

AbstractThe role of mesocarnivores in wildlife communities tends to be overlooked, which has led to a large gap in natural history knowledge. In northeastern Argentina, two mesocarnivore foxes, Cerdocyon thous and Lycalopex gymnocercus, live in sympatry in noncompetitive coexistence. Their feeding habits have been studied in many occasions; however, the studies related to trophic variability in a wider spatial scale remain scarce. In this study, we aimed to analyze how the diet of these foxes varies in three protected areas in Argentina: the Mburucuyá National Park (MNP), the “San Nicolás Portal” (SN), and the Natural Reserve Rincón de Santa María (RSM). We collected fecal samples in each area, identified them using thin layer chromatography at species level, and classified the found feeding items to obtained two parameters: occurrence and relative frequency. Furthermore, we compared the diet of the two species in each site and between sites using chi‐square and estimated trophic niche breadth and overlap between the species. We found that both species consumed different food categories, with variable frequencies across study sites. We observed that in each area, the foxes differed in diet composition. However, we found no differences between the diet compositions of C. thous and L. gymnocercus, showing high niche overlap in each site. Here, we show the plasticity of both fox species in changing their diets even with small geographical variation. Both species can coexist even with a high diet overlap, possibly explained by their noncompetitive coexistence which may occur due to the current changes in the ecosystem interactions, or partitioning may exist but on a smaller niche level.

Antibiogram, Phenotypic and Genotypic Detection of Extended-Spectrum β-lactamase Producing and Fluoroquinolone Resistant Escherichia coli Isolated from Pigs

Antimicrobial resistance in food animals is a global issue. Particularly, the dissemination of Extended Spectrum Beta-Lactamase and fluoroquinolone resistant Escherichia coli in food-producing animals posed a public health threat to the efficacy of various antibiotics. Therefore, this study aimed to detect the presence of multidrug-resistant Escherichia coli from fecal samples of pigs in Puducherry. In total, 16 faecal samples from apparently healthy pigs of different age groups were chosen for this study and subjected to detection of ESBL production and FQ resistance. E. coli was isolated from all the faecal samples (100%) and confirmed by biochemical tests and Polymerase Chain Reaction. All the E. coli isolates were found to be positive for ESBL production and fluoroquinolone resistance by phenotypic method. Out of 16 E. coli isolates, 11 (68.75%) isolates were positive for the presence of ESBL-producing genes, in which, 2 harbored bla TEM and 9 harboured bla SHV genes. In total 13/16 (81.2%) E. coli isolates harbored PMQR genes, in which 11 isolates harbored the qnrS gene and 11 isolates harbored the qnrB gene. The results showed that 9 isolates harbored both qnrS and qnrB genes in combination. Antimicrobial susceptibility test showed that the majority of the isolates exhibited a high level of resistance against Amoxycillin/clavulanic acid (100%), Cefpodoxime (100%), Cefotaxime (80%), Aztreonam (70%), Ceftazidime (70%), Enrofloxacin (70%) and Ceftriaxone (40%). The present study showed the presence of ESBL-producing and FQ-resistant E. coli in swine in the Puducherry region and their antibiogram pattern.

Evaluation of different inflammatory markers during the infection of domestic cats (Felis catus) by Cystoisospora felis (Coccidia: Apicomplexa).

Cystoisospora felis or Isospora felis is a ubiquitous apicomplexan protozoon parasite infecting domestic cats worldwide. this study aims to identify the causative agent of diarrhea in cats by determining several elevating stressors caused by these coccidian protozoans with molecular characterization. So, from January 2023 to April 2023, a total of 370 domestic cats were hospitalized at various clinics in the Cairo and Giza Governorates. Fecal samples were taken from these animals and examined by concentration floatation techniques using a saturated salt solution. The positive samples were sporulated to identify the collected oocyst. Venous blood was taken from the infected cats to evaluate the associated oxidative stress marker (lipid peroxidation products (MDA). Out of 370 examined domestic cats, 27(7.29%) were positive for C. felis. The MDA levels increased with age, and females were higher than males. DNA was extracted from fecal samples for amplification of the ITS1 gene, followed by sequencing. The ITS1 gene was amplified and showed bands at 224bp. The partial nucleotide sequence of the ITS1 gene was aligned with the reference sequences. C. felis increases the free radicals, which in turn means the animals have stress and need a schedule to treat these animals with new, safe protocol drugs that give no resistance and are highly efficient.

Syringin ameliorates dextran sulphate colitis via alteration oxidative stress, inflammation NF-κB signalling pathway and gut microbiota.

The objective of the current study was to investigate the potential effects of syringin against dextran sulphate colitis (DSS)-induced ulcerative colitis (UC) in mice. In vitro study was performed on the RAW 264.7 cells and cytokines and inflammatory level were estimated. The oxidative stress, inflammatory cytokines, apoptosis and inflammatory parameters were estimated. The mRNA expression and faecal samples were estimated in the colon tissue. Syringin treatment enhanced the body weight, colon length and reduced the disease activity index (DAI), spleen index. Syringin treatment remarkably suppressed the level of nitric oxide (NO), myeloperoxidase (MPO), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) along with alteration of antioxidant parameters. Syringin treatment also altered level of cytokines in the serum and colon tissue; inflammatory parameters viz., platelet-activating factor (PAF), cyclooxygenase-2 (COX-2), prostaglandin (PGE2), inducible nitric oxide synthetase (iNOS), nuclear factor κ-B (NF-κB); matrix metalloproteinases (MMP) level. Syringin significantly (p < 0.001) enhanced the level of nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase-1 (HO-1). Syringin remarkably altered the relative abundance of gut microbiota like Firmicutes, Bacteroidetes, F/B ratio, Verrucomicrobia and Actinobacteria. Syringin exhibited the protective effect against DSS-induced UC in mice via alteration of NF-κB signalling pathway.

Faecal sample storage without ethanol for up to 24h followed by freezing performs better than storage with ethanol for shotgun metagenomic microbiome analysis in patients with inflammatory and non-inflammatory intestinal diseases and healthy controls.

The influence of different faecal collection methods on metagenomic analyses remains under discussion, and there is no general agreement on which collection method is preferable for gut microbiome research. We compared faecal samples collected in tubes without preservatives with those containing 10mL of 96% ethanol for gut microbiome research when the timeframe from defecation to freezing at -80°C was up to 24h. We aimed to compare the collection methods on faeces from participants with inflammatory and non-inflammatory gastrointestinal disorders and healthy controls to investigate the most suitable method when considering data yield, human fraction of sequencing reads, and ease of use. We also examined the faecal sample homogeneity. Faeces collected in tubes without preservatives resulted in more sequencing reads compared to faeces collected in tubes with 96% ethanol and were also easier to handle. The human fraction of total reads in faeces collected in ethanol from participants with inflammatory bowel disease was higher than all other samples. DNA extraction and sequencing from two different locations in the same faecal sample gave similar results and showed sample homogeneity.

Baseline gut microbiome alpha diversity predicts chemotherapy-induced gastrointestinal symptoms in patients with breast cancer.

Chemotherapy frequently causes debilitating gastrointestinal symptoms, which are inadequately managed by current treatments. Recent research indicates the gut microbiome plays a role in the pathogenesis of these symptoms. The current study aimed to identify pre-chemotherapy microbiome markers that predict gastrointestinal symptom severity after breast cancer chemotherapy. Fecal samples, blood, and gastrointestinal symptom scores were collected from 59 breast cancer patients before, during, and after chemotherapy. Lower pre-chemotherapy microbiome alpha diversity and abundance of specific microbes (e.g., Faecalibacterium) predicted greater chemotherapy-induced gastrointestinal symptoms. Notably, tumor and diet characteristics were associated with lower pre-chemotherapy alpha diversity. Lower baseline alpha diversity also predicted higher chemotherapy-induced microbiome disruption, which was positively associated with diarrhea symptoms. The results indicate certain cancer patients have lower microbiome diversity before chemotherapy, which is predictive of greater chemotherapy-induced gastrointestinal symptoms and a less resilient microbiome. These patients may be strong candidates for pre-chemotherapy microbiome-directed preventative interventions (e.g., diet change).

Sex-Induced Changes in Microbial Eukaryotes and Prokaryotes in Gastrointestinal Tract of Simmental Cattle

This study investigates gender-based differences in the gut microbiota of Simmental cattle, focusing on bacterial, archaeal, and fungal communities. Fecal samples were collected and analyzed using high-throughput sequencing, with taxonomic classification performed through the SILVA and UNITE databases. Alpha and beta diversity metrics were assessed, revealing significant differences in the diversity and composition of archaeal communities between males and females. Notably, females exhibited higher alpha diversity in archaea, while beta diversity analyses indicated distinct clustering of bacterial and archaeal communities by gender. The study also identified correlations between specific microbial taxa and hematological parameters, with Treponema and Methanosphaera showing gender-specific associations that may influence cattle health and productivity. These findings highlight the importance of considering gender in microbiota-related research and suggest that gender-specific management strategies could optimize livestock performance. Future research should explore the role of sex hormones in shaping these microbial differences.

Correlation between intestinal microbiota and urolithin metabolism in a human walnut dietary intervention.

This study is to investigate the relationship between the intestinal microbiota and urine levels of the ellagic acid-derived polyphenols, the urolithins, in a cohort of subjects following a three-week walnut dietary intervention. We longitudinally collected fecal and urine samples from 39 subjects before and after walnut consumption (2oz per day for 21days). 16S RNA gene sequencing was performed on fecal DNA to study the association between microbiota composition and the levels of nine urolithin metabolites, which were measured using UHPLC/Q-TOF-MS/MS. Fecal microbial composition was found to be significantly different between pre- and post-walnut intervention (beta diversity, FDR-p = 0.018; alpha diversity, p = 0.018). Roseburia, Rothia, Parasutterella, Lachnospiraceae UCG-004, Butyricicoccus, Bilophila, Eubacterium eligens, Lachnospiraceae UCG-001, Gordonibacter, Paraprevotella, Lachnospira, Ruminococcus torques, and Sutterella were identified as the 13 most significantly enriched genera after daily intake of walnuts. We observed 26 genera that were significantly associated with 7 urolithin metabolites, with 22 genera positively correlating after walnut supplementation (FDR-p ≤ 0.05). PICRUSt analysis showed that several inferred KEGG orthologs were associated with 4 urolithin metabolites after walnut intake. In this study, we found that walnut supplementation altered urolithin metabolites, which associates with specific changes in bacterial taxa and inferred functional contents.

Gut microbiota diversity is prognostic and associated withbenefit from chemo-immunotherapy in metastatic triple-negative breast cancer.

The gut microbiota influences multiple aspects of human health and disease. Several studies have indicated an association between the gut microbiota and response to immune checkpoint inhibitors in various cancers, but there is scarce data from breast cancer. The randomized ALICE trial demonstrated improved progression-free survival (PFS) from adding the programmed cell death 1 ligand 1 (PD-L1) inhibitor atezolizumab (atezo) to immunomodulating chemotherapy (chemo) in metastatic triple-negative breast cancer (mTNBC), even for PD-L1negative disease. Herein, we investigated the microbiota composition and dynamics in the ALICE patients and their association with clinical outcome, by analyzing fecal samples collected at baseline and after 8 weeks. We applied 16S (V3-V4) rRNA sequencing to characterize the diversity and taxonomic composition. Kaplan-Meier and Cox proportional hazard models were used for time-to-event analyses. We found that high alpha diversity by Faith's phylogenetic diversity (PD) at baseline was associated with prolonged PFS in the total study population and in the atezo-chemo arm, but not in the placebo-chemo arm. Moreover, Faith's PD appeared to be predictive of benefit from atezolizumab. Patients with high Faith's PD exhibited a PFS hazard ratio of 0.34 (P = 0.018) in favor of the atezo-chemo arm, compared to 0.83 (P = 0.62) in the low Faith's PD group. Faith's PD was significantly reduced during treatment. At baseline, Bifidobacterium was significantly overrepresented in patients without clinical benefit in the atezo-chemo arm, but not in the placebo-chemo arm. These findings suggest that alpha diversity by Faith's PD should be further investigated as a prognostic and predictive biomarker in patients with mTNBC receiving chemo-immunotherapy.

Lactobacillus Helveticus Improves Controlled Cortical Impact Injury-Generated Neurological Aberrations by Remodeling of Gut-Brain Axis Mediators.

Considerable studies augured the potential of gut microbiota-based interventions in brain injury-associated complications. Based on our earlier study results, we envisaged the sex-specific neuroprotective effect of Lactobacillus helveticus by remodeling of gut-brain axis. In this study, we investigated the effect of L. helveticus on neurological complications in a mouse model of controlled cortical impact (CCI). Adult, male and female, C57BL/6 mice underwent CCI surgery and received L. helveticus treatment for six weeks. Sensorimotor function was evaluated via neurological severity score and rotarod test. Long-term effects on anxiety-like behavior and cognition were assessed using the elevated-zero maze (EZM) and novel object recognition test (NORT). Brain perilesional area, blood, colon, and fecal samples were collected post-CCI for molecular biology analysis. CCI-operated mice displayed significant neurological impairments at 1-, 3-, 5-, and 7-days post-injury (dpi) and exhibited altered behavior in EZM and NORT compared to sham-operated mice. However, these behavioral changes were ameliorated in mice receiving L. helveticus. GFAP, Iba-1, TNF-α, and IL-1β expressions and corticotrophin-releasing hormone (CRH) levels were elevated in the perilesional cortex of CCI-operated male/female mice. These elevated biomarkers and decreased BDNF levels in both male/female mice were modified by L. helveticus treatment. Additionally, L. helveticus treatment restored altered short-chain fatty acids (SCFAs) levels in fecal samples and improved intestinal integrity but did not affect decreased plasma levels of progesterone and testosterone in CCI mice. These results indicate that L. helveticus exerts beneficial effects in the CCI mouse model by mitigating inflammation and remodeling of gut microbiota-brain mediators.