Soil-based Probiotic

This study is aimed to isolate some novel probiotics from the soils of North East Himalayas. Eleven Gram positive isolates were obtained in MRS with Oxgall media from soil samples. Four of the isolates withstood the in vitro gastric juice pH 3.0 and 0.45% bile salt tolerance screening. Among these, PBT 3 showed high cell surface hydrophobicity and adhered to the Caco-2 cells. 16s rDNA gene sequences of this probiotic strains were identified as Bacillus amyloliquefaciens (accession no: JF836079). In in vivo bioefficacy evaluation by DSS-induced colitis animals, B. amyloliquefaciens significantly ameliorated the loss in body weight. Further, the treatment altered the levels of myeloperoxidase, lipoperoxides, and mucous content in the colon tissues compared with normal colon. It reduced the protein and mRNA levels of pro-inflammatory cytokines such as TNF-α and IL-1β. These biochemical findings were supported by histopathological evidences. Our study reports the use of B. amyloliquefaciens isolated from Himalayan soil as probiotic and its beneficial effect on IBD for the first time and suggests that this could be used as potential probiotics in functional foods or as a curative agent.

174. Posit Health News. 1998 Spring;(No 16):16-8. Soil-based organisms improve immune function: shift cytokine profile from TH2 to TH1. [No authors listed] AIDS: SRL 172, a protein derived from a soil-based organism, is advertised for being able to restart the immune systems of persons with allergies, lung cancer, and tuberculosis. SRL 172 also appears to shift the cytokine profile in persons with Gulf War Syndrome and Chronic Fatigue Syndrome from TH2 back to the more effective TH1 profile. One patient reports improved DTH skin responses to topical DNCB after using Nature's Biotics Soil-based Organisms (SBOs). There also appeared to be improved immunity response indicating a shift from a weaker TH2 to a TH1 cytokine profile. The patient explains his successful use of SBOs and reveals that all but 1 out of 100 other patients who have used the product for a variety of ailments cite successful outcomes. Among the ailments whose symptoms the SBOs have helped relieve are foot sores from diabetes, Chronic Fatigue Syndrome, insomnia, and genital herpes. Recent use of SBOs by HIV-infected patients has had positive results in energy and weight gain and improvements in some types of chronic conditions indicating an improvement of cell mediated immunity and antibody production.

Probiotics are known to improve gut microbiota balance, enhance food digestion, and support overall health. Among these, Bacillus species are particularly promising due to their safety, spore-forming ability, environmental resilience, and diverse enzymatic activities. However, most Bacillus probiotics used in industry are of terrestrial origin, leaving marine-derived strains largely unexplored. Utilising the untapped potential of marine microbial biomass, this study presents a multi-stage methodology for identifying and evaluating marine-derived Bacillus strains with probiotic potential. A structured screening pipeline was applied to 67 microbial isolates from the Great Barrier Reef sponges. Initial selection focused on essential probiotic characteristics, including growth, stability, safety, and survival under gastrointestinal conditions. Strains meeting these criteria were then assessed for desirable properties, including digestive enzyme production and pathogen inhibition. Using this workflow, three marine-derived Bacillus strains were identified as potential probiotics, one of which demonstrated strong antimicrobial activity against Salmonella enterica at 5 and 10 mg/mL (p < 0.01). These findings demonstrate the capability of marine-associated Bacillus as novel bioproducts with functional antimicrobial properties.

We aimed to evaluate the efficacy and safety of a spore-forming probiotic blend containing Clostridium butyricum IDCC 1301, Weizmannia coagulans IDCC 1201, and Bacillus subtilis IDCC 1101 for improving bowel function and well-being in adults with functional constipation (FC). In a randomized, double-blind, placebo-controlled trial, 78 adults with FC (Rome IV criteria) received either probiotic blend (n = 40) or placebo (n = 38) daily for 4 weeks. Primary outcomes were changes in weekly spontaneous bowel movements (WSBM) and stool form. Secondary outcomes included physical functioning scores from the 36-Item Short Form Health Survey. The probiotic blend group showed significant improvements in irritant bowel movements (p = 0.0458), incomplete evacuation (p = 0.0374), and abdominal pain before defecation (p = 0.0090). Stool consistency shifted toward normal types (Bristol types 3-4, p = 0.0176). Physical functioning improved only in the probiotic blend group (p = 0.0300). Probiotic blend effectively alleviated symptoms of FC and improved physical well-being. Trial registration: Clinical Research Information Service (CRIS), KCT0010085. Copyright: © 2026 Park et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Yeast cell wall components, being natural, biodegradable, and generally recognized as safe, offer a promising alternative to synthetic encapsulants for probiotic delivery. This study aimed to evaluate baker's yeast (Saccharomyces cerevisiae) cell wall as an encapsulant for improving the stability and gastrointestinal survivability of probiotics. Two probiotic strains with complementary functional traits were selected: Lactobacillus plantarum (a non-spore-forming lactic acid bacterium sensitive to gastric stress) and Bacillus subtilis (a spore-forming, robust probiotic widely used in feed and pharmaceutical applications). Probiotic cells (≈108-109 colony forming unit mL-1) were encapsulated within hollow yeast cell wall particles obtained via sequential acid-alkali treatment. Encapsulation efficiency, particle size, surface charge, structural integrity, and probiotic survival under simulated gastrointestinal conditions were evaluated. Scanning electron microscopy revealed a porous, honeycomb-like yeast cell wall structure (3-6 μm) facilitating probiotic encapsulation. FTIR analysis confirmed the successful encapsulation of Bacillus subtilis and Lactobacillus plantarum within the yeast cell wall matrix. Spectral changes indicated that encapsulation was driven primarily by non-covalent interactions, dominated by hydrogen bonding between yeast β-glucan hydroxyl groups and probiotic surface biomolecules. Dynamic light scattering showed a narrow and uniform size distribution of unloaded yeast cell wall (D50 = 0.63 μm; span = 0.42), while microencapsulation increased particle size, yielding a relatively uniform distributions for B. subtilis (D50 = 0.89 μm; span = 0.79) and a moderately polydisperse profile for L. plantarum (D50 = 1.67 μm, span = 1.28). Zeta potential values shifted from -16.4 ± 0.53 mV (unloaded yeast cell wall) to -32.73 ± 1.39 mV (B. subtilis) and -30.36 ± 0.42 mV (L. plantarum), indicating enhanced colloidal stability (p < 0.05). Encapsulation efficiencies were 89.6% ± 3.19% (B. subtilis) and 86.57% ± 1.50% (L. plantarum), significantly higher than their non-encapsulated counterparts (75.0% ± 2.26% and 40.6% ± 16.3%, respectively; p < 0.05). Encapsu

Bacillus subtilis has attracted attention as a robust probiotic candidate to replace antibiotic growth promoters (AGPs) in poultry, owing to its spore-forming resilience and capacity to modulate gut function. However, inclusion levels and the efficacy of multi-strain B. subtilis mixtures remain underexplored, particularly at high stocking densities, when birds are more susceptible to physiological and environmental stressors. This study evaluated the effects of incremental levels of a multi-strain B. subtilis probiotic mixture on growth performance, nutrient digestibility, physiological responses, carcass traits, and meat quality of broiler chickens reared under intensive conditions. A total of 900 one-day-old Ross 308 broiler chicks were assigned to six dietary treatments in a completely randomized design, with 36 pens (13 birds/0.75 m²) serving as experimental units. Diets comprised a basal diet without additives (PRB0), a positive control containing zinc bacitracin (POSC), and four diets supplemented with 0.3, 0.4, 0.5, or 0.6% of the B. subtilis probiotic mixture (PRB3 - PRB6). Diets were fed from the starter to the finisher phases. Probiotic supplementation influenced growth responses across the graded inclusion levels. Birds receiving the 0.5% inclusion level consistently exhibited the most favourable body weight gain and feed conversion efficiency compared with the other treatments. Probiotic supplementation enhanced the digestibility of organic matter, neutral detergent fibre, and gross energy, whereas crude protein digestibility exhibited a quadratic response. Haemato-biochemical profiles suggested improved physiological status in birds receiving mid-level probiotic inclusions (PRB3 - PRB5). Carcass yield and major meat quality traits were unaffected by dietary treatments, while minor effects on gut morphological indices were observed. Collectively, these findings suggest that a 0.5% inclusion level may be a promising probiotic supplementation strategy for broiler production under high stocking density. © 2026 The Authors.

Bacillus subtilis is a spore-forming probiotic with an established safety profile, yet its effects on aging-related phenotypes remain incompletely defined. Here, we assessed the lifespan- and healthspan-promoting activity of a novel strain, B. subtilis WTC019, using integrated invertebrate, cellular, and mammalian aging models. In Caenorhabditis elegans, B. subtilis WTC019 significantly extended lifespan, increasing median lifespan by 17.48%, 90% lifespan by 35.36%, and maximum lifespan by 19.07%, and attenuated age-associated locomotor decline. In human skin fibroblasts, B. subtilis WTC019 cell lysate reduced senescence-associated β-galactosidase activity by approximately 34% and altered cell cycle distribution consistent with delayed cellular senescence. Moreover, dietary supplementation with B. subtilis WTC019 significantly prolonged lifespan in C57BL/6J mice, with median lifespan increases of 5.97% in females and 6.05% in males. Together, these results demonstrate that B. subtilis WTC019 promotes lifespan- and healthspan-associated phenotypes across multiple aging models, supporting its potential as a probiotic candidate for healthy aging interventions.

Irritable bowel syndrome (IBS) is a chronic functional disorder characterised by abdominal pain and altered bowel habits. The most prevalent subtype is diarrhoea-predominant IBS (IBS-D). The combination of Bacillus subtilis HU58 and Heyndrickxia faecalis (formerly Bacillus coagulans) SC208 has previously exerted positive effects in people with antibiotic-associated diarrhoea and infective diarrhoea. The present multicentre study conducted in India aimed to evaluate the effectiveness and safety of the dual-strain probiotic in adults (18-65 years) with IBS-D. In this randomised, double-blind, placebo-controlled pilot study, 61 participants were recruited and assessed for changes in abdominal pain intensity (Numeric Rating Scale, NRS) and stool consistency (Bristol Stool Form Scale, BSFS) over a 4-week intervention period, with secondary outcomes including responder rates for IBS Global Assessment of Improvement (IBS-GAI) and perceived stress (Perceived Stress Scale, PSS). The probiotic group showed significantly higher overall responder rates for both abdominal pain and stool consistency ( P = 0.003) compared to the placebo group. Significant improvements were observed in abdominal pain ( P = 0.003) and stool consistency ( P = 0.035) scores in the probiotic versus placebo group from baseline to end of intervention. IBS-GAI responder rates were significantly higher among the probiotic versus placebo group ( P = 0.017) whilst perceived stress scores did not differ significantly between groups. In conclusion, supplementation with B. subtilis HU58 and H. faecalis SC208 for 4 weeks was safe and effective in improving stool consistency and abdominal pain in individuals with IBS-D, supporting its potential for symptom management in IBS-D. The trial is registered at https://ctri.nic.in/Clinicaltrials (CTRI/2022/07/044154).

Shouchella clausii is a spore-forming, Gram-positive bacterium with intrinsic antibiotic resistance and promising potential in biotherapeutics, industrial biotechnology and environmental applications. Its genetic intractability, due to a rigid cell wall and lack of natural competence, has limited its development as a microbial chassis. To facilitate its genetic transformation, a hyperosmotic electroporation protocol was optimised using cell wall weakening agents, achieving efficiencies comparable to other recalcitrant bacilli. A comprehensive and reusable genetic tool was developed centred on a temperature-sensitive E. coli-S. clausii shuttle vector (pM4B522) specifically engineered for compatibility with Golden Gate assembly. The plasmid backbone includes a spectinomycin resistance marker and an integrated red fluorescent protein reporter for transformants selection. A removable AmilCP chromoprotein cassette streamlines the assembly process by enabling blue/white screening in E. coli. This plasmid, employing a two-step pop-in/pop-out integration strategy, has demonstrated high versatility for genome editing in both S. clausii and Bacillus subtilis as evidenced by its successful use in multiple cases: (i) sequential, markerless deletions of the non-essential catabolic genes xylA and lacA in S. clausii DSM 8716, with a success rate exceeding 60%; (ii) replacement of the lacA coding sequence with a gfp coding sequence, resulting in fluorescence induction in lactose-supplemented medium; (iii) introduction of a single-base substitution generating a premature stop codon in lacA, showcasing scar-free point mutagenesis; and (iv) transfer of the system to B. subtilis 168, highlighting its broader applicability across Gram-positive bacteria. Given the precision and scarless nature of these genetic modifications, this system holds strong potential for the development of next-generation probiotics and synthetic biology applications. © 2026 The Author(s). Microbial Biotechnology published by John Wiley & Sons Ltd.

INTRODUCTION: The oral microbiota plays a fundamental role in maintaining both oral and systemic health, while dysbiosis contributes to diseases such as dental caries and periodontitis. Probiotics have gained attention as adjunctive strategies to restore microbial homeostasis. Heyndrickxia coagulans (formerly Bacillus coagulans) is a spore-forming, lactic acid-producing bacterium with documented antimicrobial, antioxidant, and immunomodulatory properties. Its resilience to environmental stressors and industrial processing makes it a promising probiotic candidate. This systematic review and meta-analysis aimed to evaluate the effects of H. coagulans on oral health outcomes. METHODS: A comprehensive search was performed across multiple databases up to September 2025 to identify randomized controlled trials (RCTs) and non-randomized studies assessing H. coagulans as a probiotic intervention for oral health. Data extraction followed PRISMA guidelines, and the risk of bias was evaluated using the RoB 2.0 and ROBINS-I tools. Meta-analysis was conducted using Stata SE® 18.5, with changes in salivary Streptococcus mutans levels as the primary outcome measure. RESULTS: Eight studies (seven RCTs and one NRSI) met the inclusion criteria. Most were conducted in India, Iran, and North Macedonia, with sample sizes ranging from 30 to 183 participants aged 5-73 years. Administration of H. coagulans, via chewable tablets, mouthwash, or food matrices, resulted in a significant reduction of salivary S. mutans counts in both children and adults compared with placebo or other probiotics. Meta-analysis of four studies demonstrated a pooled effect size of -0.99 (95%CI = -1.60/0.39; p < 0.01), although substantial heterogeneity was observed (I² = 98.2%). Additional studies reported improvements in Gingival Index, bleeding on probing, and clinical attachment levels among participants with gingivitis or periodontitis. No significant adverse events were reported. CONCLUSION: H. coagulans appears to exert beneficial effects on oral health by reducing cariogenic bacterial load and improving periodontal parameters, supporting its potential use as an adjunct in caries prevention and gingival

The use of probiotics in aquaculture has emerged as a sustainable strategy for enhancing fish health and reducing the use of antibiotics, thereby serving the cause of One Health. In the present study, we employed a novel, distinctive, rigorous, and multi-tiered plate-based screening methodology to isolate spore-forming probiotic bacterial strains from fish market drain water for aquaculture applications. Based on their non-pathogenicity, potential for extracellular enzyme production, bile salt tolerance, and biofilm formation capabilities, three isolates, FM1, FM2, and FM4, with positive performance for all tested assays, were chosen and comprehensively characterized. Strain FM2, identified based on 16S rRNA gene sequence homology as Bacillus sp., was highly effective, exhibiting high sporulation efficiency (91%) and yield (9.3 × 108 CFU /mL). It also had broad-spectrum antimicrobial activity against eight common fish pathogens, with high biofilm-formation inhibition, and preformed biofilm-disruption potential. The strain further demonstrated excellent adhesive capabilities (97% auto-aggregation at 24 h, significant co-aggregation with pathogens, and high cell surface hydrophobicity), robust antioxidant activity, and notable exopolysaccharide production. It was susceptible to key antibiotics, including gentamicin, imipenem, amikacin, and ciprofloxacin, ensuring biosafety. Its spores can withstand environmental and gastrointestinal stress conditions, such as heat (110 °C), UV light, lysozyme, and bile salts, and survive in simulated gastric and intestinal fluids. Thus, FM2 is a promising, safe, and multi-functional probiotic for aquaculture, capable of improving fish gut health, inhibiting pathogens, and surviving under challenging environmental conditions, and has enormous potential for commercialisation in aquaculture. © 2026. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

This review focuses on the application of Bacillus-based probiotics in livestock production, emphasizing their potential to enhance animal health, growth performance, and welfare through modulation of the gut microbiota, immune function, and nutrient absorption. Bacillus probiotics, particularly spore-forming strains, offer advantages such as stability during feed processing or the ability to survive gastrointestinal conditions, germinate and produce beneficial metabolites in the intestine. The mechanisms by which Bacillus probiotics exert their effects include influencing microbial communities, producing bioactive compounds, and strengthening gut barrier integrity, which together lead to improved digestive health and resistance to pathogens. Additionally, the challenges in standardizing their effects and identifying reliable biomarkers for evaluating probiotic efficacy or the axes with other organs are highlighted. Overall, Bacillus probiotics are seen as promising, sustainable alternatives to antibiotics with significant potential for future research to optimize their use and understand their mechanisms within the context of animal production and health.

Bacillus licheniformis is a spore-forming bacterium with probiotic, environmental, and industrial applications. Many wild strains with diverse functions have been described in recent years. Nevertheless, the lack of efficient and universal genetic manipulation tools hinders the study and engineering of these strains. Here, a versatile and simple genetic manipulation toolkit is established for B. licheniformis. The cornerstone of this toolkit is a conjugative DNA transfer system. This system could effectively transfer temperature-sensitive plasmid pTSMK into all ten tested B. licheniformis strains, with efficiencies ranging from 10-5 to 10-3. Based on this DNA transfer system, the tools for maker-free knockout and knock-in, CRISPRi, as well as transposon mutagenesis, were built. A transposition frequency of 7.68 × 10-3 was observed. The toolkit developed in this study fulfills most tasks in the engineering of this species and will promote the basic and applied research of B. licheniformis.

OBJECTIVE: Bacillus coagulans is a spore-forming probiotic known for its resilience and metabolic activity, both of which are desirable in promoting gut health and oxidative balance. Nonetheless, the beneficial effects of B. coagulans-fermented bedding (BFB) on raising native Chinese chicken breed farming remain largely unknown. This study was conducted to evaluate BFB supplementation on growth performance, meat quality and gut health in Jiuyuan Black chicken. METHODS: A total of 120 male chicks were allocated to the control (CON, using traditional litter) and BFB groups with four replicates per group containing fifteen birds. The chickens were monitored for 70 days; growth performance was evaluated on days 35 and 70, while meat quality, intestinal integrity, antioxidant capacity, and animal welfare were evaluated on day 70. RESULTS: The results showed that, after 70 days, the chickens in the BFB group exhibited significantly higher average daily gain, lower feed conversion ratio, and increased semi-eviscerated yield and intramuscular fat content compared to the CON group (P < 0.05). Breast muscle from the BFB group showed enhanced flavor and juiciness than the CON group (P < 0.05). Furthermore, the histological analysis of the jejunum demonstrated increased villus height-to-crypt depth ratio, alongside upregulated expression of tight junction proteins (Claudin-1 and ZO-1) (P < 0.05). Additionally, total antioxidant capacity, catalase, and superoxide dismutase activities were increased, with reduced malondialdehyde levels in the serum and jejunal tissue (P < 0.05). Furthermore, BFB improved Jiuyuan Black chickens feather coverage (P < 0.05). CONCLUSION: This study indicated that BFB treatment was a good source of reducing oxidative stress in broilers by improving gut health, antioxidant capacity and meat quality which may provide an essential proof for the practical application to enhance growth performance without causing welfare issues in poultry.

BACKGROUND: The immune system and the gastrointestinal tract are intricately linked. The intestinal barrier, microbiome, and immune system are in constant communication, shaping immune responses and maintaining homeostasis. Imbalances in the gut microbiome can affect the intestinal barrier and increase susceptibility to infections, along with a decline in immune function (both innate and adaptive immunity). Maintaining optimal immune function is crucial for protecting against infections and supporting overall health, particularly in populations that may be more vulnerable to seasonal respiratory and gastrointestinal infections. Probiotics, particularly spore-forming strains, demonstrate potential for improving natural killer (NK) cell function and mucosal immunity through gut-associated lymphoid tissue interactions. This study evaluated the immunomodulatory effects of Bacillus coagulans SNZ 1969® in adults. METHODS: This randomized, double-blind, placebo-controlled clinical trial enrolled adults aged 60-65 years who were susceptible to seasonal infections. Participants were randomized 1:1 to receive either B. coagulans SNZ 1969® (2 billion CFU/day) or placebo for 12 weeks. Primary endpoints included NK cell activity, absolute NK cell count (CD3⁻/CD16⁺/CD56⁺), and immunoglobulin levels (serum IgM, IgG, and IgA and salivary IgA). Secondary outcomes assessed respiratory and gastrointestinal infection incidence, inflammatory markers (C-reactive protein), and safety parameters. RESULTS: Of 60 enrolled participants, 50 completed the study (25 per group). B. coagulans SNZ 1969® significantly enhanced NK cell activity compared to placebo, with a net increase of 42.07% between groups (44.59% versus 2.52% increase from baseline; p = 0.0002). NK cell activity improvements were consistent across both genders in exploratory subgroup analyses (limited by small female n = 7-8 per arm; males: 36.75% versus 2.52%; p = 0.00004; females: 63.64% versus 2.71%; p = 0.01155). Significant improvements were observed in serum IgA (25.00% versus 2.30% change; p = 0.0016) and salivary IgA (27.70% versus 0.60% change; p = 0.0002). No significant changes occurred in absolute NK cell counts,

Atherosclerosis is a chronic inflammatory disease characterized by lipid accumulation and immune dysregulation, including expansion of pro-inflammatory monocytes and effector T cells, alongside reduced regulatory T cells (Tregs). Bacillus subtilis natto, a spore-forming probiotic, has shown anti-atherosclerotic effects, though its systemic immunomodulatory mechanisms remain unclear. In this study, we employed an AAV-mPCSK9-induced murine atherosclerotic model to investigate the effects of daily B. subtilis natto NTU-18 administration over 16 weeks. High-dimensional flow cytometry using two 13-marker panels enabled longitudinal profiling of 18 peripheral immune cell subsets across lymphoid and myeloid compartments. While no significant changes in serum cholesterol and mild decrease of body weight were observed, B. subtilis natto NTU-18-treated mice presented a significant reduction in aortic lesion area compared to PBS-treated controls. Immune profiling revealed a transient expansion of peripheral myeloid cells and CD44⁺ trained CD8⁺ T cells, followed by increased frequencies of naïve CD8⁺ T cells and reduced central/effector memory subsets at longer time point treatment. In the CD4⁺ T cell compartment, a transient increase in trained cells was accompanied by a sustained enrichment of CD25⁺CD4⁺ Tregs throughout the daily B. subtilis natto NTU-18 treatment. In contrast, no significant differences were observed in Ly6C⁻ or Ly6C⁺ monocytes, neutrophils, or eosinophils. These findings suggest that B. subtilis natto NTU-18 attenuates atherosclerosis progression not through lipid lowering or broad myeloid modulation, but via targeted reprogramming of peripheral T cell responses. This work provides mechanistic insight into the immunotherapeutic potential of B. subtilis natto NTU-18 in atherosclerosis prevention and treatment. KEY POINTS: • B. subtilis natto NTU-18 significantly reduces aortic plaque burden in atherosclerotic mice without affecting serum cholesterol levels. • B. subtilis natto NTU-18 induces transient immune remodeling, marked by early expansion of trained CD8⁺ and CD4⁺ T cells, followed by increased naïve and regulatory T cells. • The atheroprotective e

The spore-forming capacity of Bacillus spp. enables environmental persistence and stable product formulations, yet the interactions of environmental Bacillus spores and vegetative cells with the human immune system are not fully understood. We investigated the immunostimulatory potential of seven environmental Bacillus isolates (B. subtilis, B. velezensis, B. licheniformis, B. pumilus) and optimized their inactivation methods to preserve surface integrity and immunostimulatory activity. Inactivation was evaluated using heat/UV-C and 10-100% formalin, followed by scanning electron microscopy and human reporter cell assays (THP1-Dual monocytes, HEK-Blue hTLR2/6 and hTLR4). Heat/UV-C treatment retained Bacillus morphology and enhanced immune activation compared to 10-100% formalin treatment. Spores and vegetative cells activated key immune transcription factors (nuclear factor kappa B and interferon regulatory factors) in human monocytes, with spores inducing 1.5- to 8-fold lower responses compared to vegetative cells, and strain- and species-dependent effects observed. All tested Bacillus isolates significantly activated Toll-like receptor TLR2/6, but not TLR4. Genome analysis identified Bacillus spore envelope components (SpsAEIK, CotBCGQ) that may influence differences in immune responses. Our findings improve understanding of Bacillus-human immune interactions, revealing strain- and species-specific immunostimulatory activity. These results support further exploration of Bacillus isolates for immune-related and environmental applications. © 2025. The Author(s).

Bacillus coagulans GBI-30, 6086, is a lactic acid-producing, spore-forming bacteria with diverse characteristics that lend it resiliency through commercial pet food processing, storage, and chemical and enzymatic digestion through the gastrointestinal tract. The objective of this study was to determine the effects of B. coagulans on dietary apparent total tract nutrient digestibility (ATTD) and the hematology, immunoglobulin concentrations and fecal characteristics, metabolites, and microbiota populations of healthy adult dogs. Adult English Pointer dogs (n = 12; age = 5.9 ± 2.5 yr; body weight = 26.6 ± 6.1 kg) were fed the same diet but supplemented with B. coagulans or a placebo via gelatin capsules in a replicated 3 × 3 Latin square design. Capsules were administered daily before each feeding, with the following treatments tested: (1) basal diet + placebo (control; 250 mg maltodextrin); (2) basal diet + B. coagulans [low dose; 5 × 108 colony-forming units (CFU)/d]; and (3) basal diet + B. coagulans (high dose; 2.5 × 109 CFU/d). Fecal, blood, nasal, and pinnae samples were collected after a 22-d adaptation phase in each period. Data were analyzed using the Mixed Models procedure of SAS 9.4, with P < 0.05 being significant and P < 0.10 being trends. Bacillus coagulans supplementation did not affect ATTD, food intake, fecal metabolites, immunoglobulin concentrations, or hematology, but did lower fecal scores (P < 0.05; firmer stool). Using qPCR, fecal Faecalibacterium spp. abundance was greater (P < 0.05) and fecal Bacteroides spp., Bifidobacterium spp., and Ruminococcus gnavus abundances tended to be greater (P < 0.10) in dogs fed the low B. coagulans dose than those fed the placebo. Overall, our results demonstrate that this bacterial species can be safely supplemented to healthy dogs without a negative impact on nutrient digestibility or canine health parameters, and supplementation with the low dose treatment may help maintain fecal consistency and positively influence gastrointestinal microbiota. Plain Language Summary: Bacillus coagulans GBI-30, 6086, is a commercially available, lactic-acid-producing, spore-forming bacteria that exhibits probiotic charac

The spore-forming probiotic Bacillus coagulans JBI-YZ6.3 interacts with the gut epithelium via its secreted metabolites as well as its cell walls, engaging pattern-recognition receptors on the epithelium. We evaluated its effects on human T84 gut epithelial cells using in vitro co-cultures, comparing metabolically active germinated spores to the isolated metabolite fraction and cell wall fraction under unstressed versus inflamed conditions. Germinated spores affected epithelial communication via chemokines interleukin-8, interferon gamma-induced protein-10, and macrophage inflammatory protein-1 alpha and beta after 2 and 24 h of co-culture. Non-linear dose responses confirmed that bacterial density affected the epigenetic state of the epithelial cells. In contrast, the cell wall fraction increased cytokine and chemokine levels under both normal and inflamed conditions, demonstrating that the intact bacterium had anti-inflammatory properties, regulating pro-inflammatory signals from its cell walls. During recovery from mechanical wounding, germinated spores accelerated healing, both in the absence and presence of LPS-induced inflammation; both the metabolite and cell wall fractions contributed to this effect. The release of zonulin, a regulator of tight junction integrity, was reduced by germinated spores after 2 h. These findings suggest that B. coagulans JBI-YZ6.3 modulates epithelial chemokine signaling, supports barrier integrity, and enhances epithelial resilience, highlighting its potential as an efficacious multi-faceted probiotic for gut health.

Antibiotics are among the most prescribed medicines in children globally. Antibiotic-associated diarrhoea (AAD) and associated abdominal pain are common side effects, caused by alterations to the intestinal microbiota composition. Most research on probiotic interventions involves prophylactic use of Saccharomyces, lactobacilli or bifidobacteria. Less is known about spore-forming strains administered after AAD onset. Bacillus subtilis HU58™ was found to improve AAD symptoms in adults. This randomised, double-blind, placebo-controlled trial in India investigated the effects of a dietary supplement containing B. subtilis HU58 (2 × 109 cfu/day) versus placebo for seven days on diarrhoea duration (Bristol Stool Scale, BSS), abdominal pain intensity (Visual Analogue Scale, VAS), and overall gastrointestinal wellbeing (adapted Gastrointestinal Restoration Questionnaire, GIRQ; Physician Global Assessment, PGA) in 68 children (1-12 years) with AAD. Between-group comparison of data from each timepoint was conducted for the intention to treat population (Chi-squared test of independence for distribution of BSS types, Wilcoxon rank sum test for mean BSS types, mixed model for repeated measures for VAS, GIRQ and PGA scores). By day 3, 93.5% of the probiotic group had normal BSS types, versus 22.6% in the placebo group ( P < 0.001). Almost all subjects in both groups had normal stool types by day 7. Significantly greater decreases in abdominal pain VAS scores were observed among those receiving the probiotic versus placebo at day 3 (-7.4 [SE 0.5] versus -1.9 [SE 0.3], P < 0.001) and day 7 (-9.1 [SE 0.3] versus -8.5 [SE 0.2], P < 0.001). Greater improvements for both adapted GIRQ and PGA scores assessing GI wellbeing were observed in the probiotic group compared to the placebo group at days 3 and 7 (all P < 0.001). Spore-forming B. subtilis HU58 administered to children after onset of AAD may lead to faster resolution of diarrhoea and associated abdominal pain. Further research with daily clinical assessment and faecal microbiome analysis is warranted. The trial is registered at https://ctri.nic.in/Clinicaltrials (CTRI/2022/02/040138).

BACKGROUND AND AIM: The pursuit of sustainable alternatives to antibiotic growth promoters has intensified interest in spore-forming probiotics with fiber-degrading capabilities. This study aimed to isolate, characterize, and evaluate the safety and functional properties of Bacillus spp. from native Thai swine, focusing on strains with probiotic potential and enzymatic activity for application in livestock nutrition. MATERIALS AND METHODS: Spore-forming Bacillus isolates were obtained from fecal samples of backyard-raised native pigs. Isolates were screened for acid and bile tolerance, autoaggregation, hydrophobicity, biofilm formation, adhesion to Caco-2 cells, antimicrobial activity, and co-aggregation with pathogens. Enzyme production (cellulase, xylanase, and pectinase), hemolytic activity, and antibiotic susceptibility were also assessed. The most promising strain, Bacillus amyloliquefaciens NL1.2, was subjected to in vivo safety and efficacy evaluations in a mouse model, including assessments of toxicity, histopathology, secretory immunoglobulin A (IgA) levels, and gut microbiome modulation through full-length 16S ribosomal RNA sequencing. RESULTS: B. amyloliquefaciens NL1.2 exhibited robust probiotic traits including high acid (115.05%) and bile (75.16%) tolerance, strong autoaggregation (65.99%), moderate hydrophobicity (34.13%), and effective adhesion (2.0%) to intestinal epithelial cells. It produced fiber-degrading enzymes (cellulase: 0.015 U/mL; xylanase: 0.522 U/mL; and pectinase: 0.374 U/mL) showed antimicrobial activity against Enterohemorrhagic Escherichia coli, Enteropathogenic E. coli, and Salmonella Typhimurium, and was non-hemolytic and antibiotic-sensitive. In vivo, NL1.2 induced no adverse effects and significantly elevated intestinal secretory IgA levels (p < 0.05). Microbiome analysis revealed enrichment of beneficial taxa (e.g., Bacteroidetes and Barnesiella) and reduction of potentially pathogenic taxa (e.g., Helicobacter and Deferribacteres). CONCLUSION: B. amyloliquefaciens NL1.2 is a safe, multifunctional probiotic with fiber-degrading, immunomodulatory, and gut microbiota-modulating properties. Its origin from native swine and broad

Spore-forming Bacilli have been explored due to their potential biotechnological features and applications in human health and functional food research. This study focuses on the genetic and phenotypical characterization of the functional probiotic properties of Bacillus velezensis P45, a strain isolated from fish intestines. B. velezensis P45 exhibited antimicrobial activity against Gram-positive and Gram-negative pathogens and demonstrated strong autoaggregation and biofilm formation properties in vitro. The strain also showed tolerance to gastrointestinal conditions and ability to metabolize and adhere to mucin. In silico analysis confirmed the absence of virulence factors and antibiotic resistance genes, reinforcing its safety as a probiotic candidate. Genome mining revealed the presence of genes related to adhesion, such as fibronectin-binding protein and enolases, and for the synthesis of secondary metabolites, including the antimicrobial lipopeptides fengycin, surfactin, and bacillibactin. In addition, phylogenetic comparison using the yloA (rqcH) gene associated with gut adhesion clustered strain P45 with other probiotic Bacillus and B. velezensis strains, while separating it from pathogenic bacteria. Thus, the strain B. velezensis P45 could be a valuable candidate as a probiotic due to its functional properties and safety.

Sulfoquinovose, a sulfonated sugar derived from the thylakoid membrane lipid sulfoquinovosyl diacylglycerol, is abundant in photosynthetic organisms and plays a key role in global sulfur cycling. Its degradation in nature is mediated by specialized bacteria, many of which rely on the enzyme sulfoquinovosidase (YihQ) to release sulfoquinovose from sulfoquinovosyl (diacyl)glycerol. Despite its ecological importance, the diversity and functional roles of sulfoquinovose-degrading microorganisms remain poorly characterized in natural environments. Here, we developed a yihQ-targeted amplicon sequencing approach to investigate the richness and distribution of SQ-degrading bacteria across selected environments. We revealed high richness of yihQ-containing microorganisms in the analyzed cow rumen samples, far exceeding that observed in human and mouse gut microbiomes, suggesting an important role of sulfoquinovose metabolism in ruminant digestion. Anoxic microcosm experiments with sulfoquinovose-amended rumen fluid revealed cooperative microbial degradation of sulfoquinovose to sulfide via isethionate cross-feeding. Amplicon sequencing and genome-resolved metagenomics and metatranscriptomics identified yet undescribed and uncultured sulfoquinovose-degrading taxa. Members of Caproiciproducens (Acutalibacteraceae), Candidatus Limivicinus (Oscillospiraceae), and Sphaerochaetaceae transcribed the isethionate-producing sulfo-transketolase pathway, whereas isethionate was likely respired by a Candidatus Mailhella bacterium (Desulfovibrionaceae). This study presents a functional gene-based assay for tracking environmental yihQ richness, highlights sulfoquinovose degradation as a central metabolic process in the cow rumen, describes previously unknown sulfoquinovose-metabolizing bacteria, and advances understanding of sulfur physiology in complex microbial communities. © The Author(s) 2026. Published by Oxford University Press on behalf of the International Society for Microbial Ecology.

The microbiome, also considered the hidden organ, is a fundamental ecosystem directly associated with the disease and health status of the human body. With the availability of high-throughput DNA sequencing technologies, a growing number of studies from clinical and experimental (observation and intervention) samples are constantly revealing new findings on the relationship between human organs and their microbiomes. In such a context, diet and nutrition are among the key factors influencing microbiome composition, richness, and functional behavior. In this review, we illustrate how microbiome-related data and associated metadata are in recent times scattered across primary and specialized databases with different levels of curation, annotation, and standardization, limiting, to some extent, the possibility of deep data discovery, reuse, alignment, and harmonization. Therefore, we describe the way Findable, Accessible, Interoperable, and Reusable (FAIR) data principles would enhance the onset of novel scientific hypotheses and potential microbiome-targeted therapies by improving the standardization policies in data sources. Accordingly, using advanced semantic classification and data mining technologies based on suitable and comprehensive ontologies, annotations of studies present in source databases or in scientific literature would further improve the data and metadata enrichment, integration and alignment relevant to microbiome data associated with health, disease and nutrition.

Hepatic encephalopathy (HE), a severe neurological complication of liver dysfunction, has long been regarded as a clinical issue confined to liver disease. However, recent clinical observations and basic research have revealed complex pathophysiological connections between HE and Parkinson's disease (PD), 2 seemingly independent conditions. Patients with HE often exhibit irreversible extrapyramidal symptoms that closely resemble the motor disorders of PD; meanwhile, epidemiological studies suggest that individuals with liver disease-particularly non-alcoholic fatty liver disease (NAFLD)-may face an increased risk of developing PD. From the perspective of the gut-liver-brain axis, this study systematically explores the molecular mechanisms linking HE and PD, proposing a core hypothesis: HE creates a unique "neurotoxic soil" through ammonia toxicity, systemic neuroinflammation, and gut-liver-brain axis dysfunction. This soil may trigger PD in susceptible individuals, accelerate subclinical PD progression, or mimic PD-like pathology. The study analyzes in depth the direct regulatory role of ammonia in α-synuclein (α-syn) aggregation, the impact of liver disease-driven neuroinflammation on microglial activation and α-syn propagation, and the hypothesis of liver-derived α-syn transmission via the gut-liver-brain axis. It further discusses synergistic mechanisms such as manganese deposition, neurotransmitter imbalance, and gut microbiota metabolites. Based on these mechanisms, the study prospects translational medical applications, including the development of diagnostic biomarkers and novel therapeutic strategies such as "ammonia clearance plus" and gut-liver-brain axis targeting. This work provides new insights into how environmental metabolic factors contribute to neurodegenerative diseases and offers a theoretical basis for the combined prevention and treatment of HE and PD.

Microplastics (MPs) derived from degraded agricultural mulching films frequently co-occur with arsenic in cropland soils, yet their joint toxic effects on soil fauna remain insufficiently understood. This study compared conventional non-biodegradable microplastics (polyethylene, PE) with biodegradable microplastics (polylactic acid, PLA; and polybutylene adipate-co-terephthalate, PBAT) co-exposed with arsenate (As(V)) in a 28-day soil exposure experiment using earthworm Eisenia fetida as model organism. Results showed that microplastic abundance in the earthworm gut increased over time, whereas co-exposure consistently reduced arsenic bioaccumulation in tissues, with stronger reductions observed for PLA and PBAT than for PE. Across multiple physiological and oxidative stress-related biomarkers, As(V) induced growth inhibition and oxidative injury, while the presence of microplastics generally mitigated these adverse responses. Integrated Biomarker Response (IBR) analysis indicated lower overall stress in the combined treatments than in the corresponding single As(V) treatments, and interaction assessment based on the IBR-Effect Addition Index (EAI) model further suggested antagonistic effects between As(V) and all three microplastics. Overall, biodegradable microplastics (PLA MPs and PBAT MPs) exhibited a greater capacity to attenuate arsenic toxicity than conventional PE MPs. These findings demonstrate polymer-dependent interactions in arsenic-microplastic co-contamination and provide a scientific basis for ecological risk assessment of conventional and biodegradable mulching films in agricultural soils.

As global water scarcity intensifies, water reuse has become a critical alternative source for agricultural irrigation, ecological recharge, and domestic use. However, considerable debate exists regarding the potential ecological risks posed by reclaimed water, particularly its impact on soil ecosystems. This study investigates the effects of reclaimed water at varying concentrations on physiological and biochemical responses, as well as gut microbiota and metabolites of earthworm Eisenia fetida. The results demonstrated that reclaimed water induced oxidative stress and damage to the gut microstructure and digestive function in earthworms. 16S rRNA sequencing and metabolomics analyses further revealed that reclaimed water exposure resulted in dysbiosis of earthworm gut microbiota, characterized by an increased abundance of pathogens and degradation-associated bacteria, and significant alterations in metabolites associated with pyrimidine and purine pathways were observed. Furthermore, reclaimed water led to reproductive toxicity in earthworms. By integrating physiological indicators and analysis of cross-talk patterns among the effects, we concluded that exposure to reclaimed water instigates systemic toxicity in earthworms. These findings demonstrate that reclaimed water induces toxic effects in soil-dwelling organisms and potentially impair the reproductive success of subsequent generations, underscoring the need for attention to the ecological risks associated with reclaimed water application in terrestrial environments.

The use of pesticides in agriculture can affect the biodiversity of soil ecosystems and interfere with the ecological services they provide. The gut microbiome of earthworms serves as a bioindicator for changes in diversity within one of the most representative groups of soil macrofauna. In this study, we defined a gradient of soil and pesticide use regimes within a horticultural area, including conventional management, good agricultural practices, organic production and forest. Earthworms (A. gracilis) from these four conditions were sampled, and their microbiomes were assessed by sequencing the V4 region from the 16S rRNA gene. Comparison of ASVs indicated the presence of 142 bacterial genera among all the samples, with Actinobacteriota, Proteobacteria, Firmicutes and Bacteroidota being dominant phyla. Simpson's diversity index revealed diminished biodiversity in the gut microbiomes of earthworms from conventionally managed soil, correlating negatively with the presence of the insecticide chlorpyrifos. Furthermore, a seasonal shift in the dominance of bacterial taxa was observed between dry and rainy seasons. These shifts were evident in the gut microbiome of organisms from organically managed and good practices soils but not in the conventional site. Our results demonstrate that earthworm's gut microbiome serves as a responsive bioindicator for biodiversity changes in agricultural landscapes, suggesting that key features of the microbial community can be altered by pesticide exposure.

Conjugation accelerates bacterial evolution by enabling bacteria to acquire genes horizontally from their neighbors. Plasmid donors must physically encounter and connect with recipients to allow plasmid transfer, and different environments are characterized by vastly different encounter rates between cells, based on mechanisms ranging from simple diffusion to fluid flow. However, how the environment affects the conjugation rate by setting the encounter rate has been largely neglected, mostly because existing experimental setups do not allow for direct control over cell encounters. Here, we describe the results of conjugation experiments in Escherichia coli in which we systematically varied the magnitude of shear flow using a cone-and-plate rheometer to control the encounter rate. We found that the conjugation rate increases with shear until it peaks at an optimal shear rate ([Formula: see text]), reaching a conjugation rate fivefold higher than the baseline set by diffusion-driven encounters. This optimum marks the transition from a regime in which shear promotes conjugation by increasing the rate of cell-cell encounters to a regime in which shear disrupts conjugation. Regions of high fluid shear are widespread in aquatic systems, in the gut of host organisms, and in soil, and our results indicate that these regions could be hot spots of bacterial conjugation in the environment.

The widespread use of plastic agricultural films necessitates a thorough evaluation of environmental risks posed by soil microplastics (MPs). While the intestinal tract is a critical site for MP interactions in soil organisms, current research predominantly focuses on overall physiological responses, overlooking organ-specific toxic mechanisms. To address this gap, we exposed earthworms (Eisenia fetida) to polyethylene (PE) and biodegradable polylactic acid (PLA) MPs sourced from agricultural films at an environmentally realistic concentration of 1.0 g/kg. Incorporating natural earthworm mobility, we designed two exposure scenarios: migration from clean to contaminated soil (scenario A) and vice versa (scenario B). Machine learning-driven image analysis and phenotypic profiling revealed that PE induced more severe intestinal lesions than PLA, adversely affecting intestinal immune functions. Furthermore, PE resulted in greater oxidative damage and significantly activated immune proteins such as melanin and antimicrobial peptides through reprograming immune-related gene and protein pathways. Conversely, PLA predominantly disrupted intestinal digestive and absorptive functions, though the gut microbial community partially mitigated damage through structural and compositional adaptation. Compared with scenario A, earthworms in scenario B exhibited reduced tissue damage, enhanced digestive enzyme activity, and upregulated energy-related metabolites and cell proliferation genes, indicating partial recovery from MP-induced intestinal dysfunction. These findings elucidate the distinct toxicity mechanisms of conventional and biodegradable agricultural MPs on soil organisms, while the scenario-based approach advances risk assessment by aligning experimental design with real-world ecological behaviors.

Benzophenone-type ultraviolet filters recently received significant attention to overlook regulatory agencies' safety potential due to their toxicological implications on humans and the environment. The present study has been carried out to explore the toxicity of Benzophenone-3 (BP-3) in earthworm Eisenia fetida. Low-level long-term exposure defiles earthworm health through elevated ROS and its detrimental impact on reproductive organs and reproduction. Based on KEGG and GO analysis, global transcriptomics reveals differentially expressed gene transcripts affecting key signaling pathways. Further validation by q-PCR showed significant upregulated expression of genes involved in stress (CuZn-SOD, CAT), metabolism (GST), reproduction and gametogenesis (ANN and Piwi-2), and endocrine (EcR) functions. Interestingly, lower concentrations of BP-3 are biologically effective in exhibiting a non-linear concentration-response pattern towards the expression of reproduction and endocrine function genes. In addition, BP-3, through soil exposure, significantly alters the gut microbiome by inducing changes in bacterial diversity, while fungal diversity remains unaffected. Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes have significantly affected phyla, whereas Ascomycota and Basidiomycota remain dominant, suggesting their potential role in metabolizing or tolerating the BP-3 contamination. The findings highlight the molecular consequences of BP-3 exposure in earthworms and indicate the broader environmental impacts of benzophenone-type organic UV filters on terrestrial biota. The information could also be helpful for chemical risk assessment in soil ecotoxicology.

Similar to many other organisms, insects like Drosophila melanogaster, Hypothenemus hampei, and Cockroaches harbor diverse bacterial communities in their gastrointestinal systems. These bacteria, along with other microorganisms like fungi and archaea, are essential to the physiology of their insect hosts, forming intricate symbiotic relationships. These gut-associated microorganisms contribute to various vital functions, including digestion, nutrient absorption, immune regulation, and behavioral modulation. Notably, gut microbiota facilitates the breakdown of complex plant materials, synthesizes essential vitamins and amino acids, and detoxifies harmful substances, including pesticides. Furthermore, these microorganisms are integral to modulating host immune responses and enhancing disease resistance. This review examines the multifaceted roles of gut microbiota in insect physiology, with particular emphasis on their contributions to digestion, detoxification, reproduction, and environmental adaptability. The potential applications of gut microbiota in integrated pest management (IPM) are also explored. Understanding the microbial dynamics within insect pest species opens new avenues for pest control, including developing microbial biocontrol agents, microbial modifications to reduce pesticide resistance, and implementing microbiome-based genetic strategies. In particular, manipulating gut microbiota presents a promising approach to pest management, offering a sustainable and eco-friendly alternative to conventional chemical pesticides. © 2025. The Author(s), under exclusive licence to Springer Nature B.V.

BACKGROUND: Global warming is redrawing the map for invasive species, spotlighting the globally harmful giant African snail as a major ecological disruptor and public health threat. Known for harboring extensive antibiotic resistance genes (ARGs) and human pathogens, it remains uncertain whether global warming exacerbates these associated health risks. METHODS: We use phenotype-based single-cell Raman with D2O labeling (Raman-D2O) and genotype-based metagenomic sequencing to investigate whether soil warming increases active antibiotic-resistant bacteria (ARBs) in the gut microbiome of giant African snails. RESULTS: We show a significant increase in beta-lactam phenotypic resistance of active ARBs with rising soil temperatures, mirrored by a surge in beta-lactamase genes such as SHV, TEM, OCH, OKP, and LEN subtypes. Through a correlation analysis between the abundance of phenotypically active ARBs and genotypically ARG-carrying gut microbes, we identify species that contribute to the increased activity of antibiotic resistome under soil warming. Among 299 high-quality ARG-carrying metagenome-assembled genomes (MAGs), we further revealed that the soil warming enhances the abundance of "supercarriers" including human pathogens with multiple ARGs and virulence factors. Furthermore, we identified elevated biosynthetic gene clusters (BGCs) within these ARG-carrying MAGs, with a third encoding at least one BGC. This suggests a link between active ARBs and secondary metabolism, enhancing the environmental adaptability and competitive advantage of these organisms in warmer environments. CONCLUSIONS: The study underscores the complex interactions between soil warming and antibiotic resistance in the gut microbiome of the giant African snail, highlighting a potential escalation in environmental health risks due to global warming. These findings emphasize the urgent need for integrated environmental and health strategies to manage the rising threat of antibiotic resistance in the context of global climate change. Video Abstract. © 2025. The Author(s).

Microplastics (MPs) are continuously found in soil and water environments. Within aquatic ecosystems, filter-feeding organisms are unable to discriminate MPs from food particles while fish may intentionally ingest MPs by mistaking them for prey. In both cases, MPs can accumulate in tissues with subsequent implications for human and environmental health. The modes of action of MPs are still not fully understood and hence the toxicological effects of these pollutants cannot be fully evaluated. This study aims to improve our understanding of the modes of action and toxicological effects of MPs using a multimodal approach. In the present study, Daphnia magna was deployed as a model to investigate the acute effects of MPs by exposing D. magna specimens for 24 h to fluorophore-coated polyethylene MPs. A multimodal approach, combining fluorescence imaging and mass spectrometry imaging (MSI), was employed to assess the implications of MPs exposures. Fluorescent microscopy revealed a significant accumulation of MPs in the gut of D. magna after acute exposure. Secondary ion mass spectrometry (SIMS) and matrix-assisted laser desorption/ionization (MALDI) imaging were used to study the distribution and potential metabolic effects in exposed D. magna. ToF-SIMS revealed specific fragmentation patterns for polyethylene MPs, with the m/z 43 ion being the most suitable for identifying polyethylene MPs in biological tissue samples. MALDI-MSI showed specific ion types for the eye, gut, optical ganglion, first antennae, and egg tissues of D. magna. MSI data revealed minor alterations in specific regions of D. magna, such as eggs and gut, of D. magna after MPs exposure. The local changes were mainly found in the nucleotide and lipid metabolism within the eggs. In the gut, changes between control and MPs-exposed groups were potentially linked to plastic additives. Overall, the results of this work underline the potential of multimodal approaches based on MSI to study challenging pollutants, such as MPs, and their interactions with tissues.

OBJECTIVES: We report a unique case of an 88-year-old man presenting with symptoms consistent with a urinary tract infection, whose diagnostic evaluation led to the identification of a previously unrecognized motile flagellated protozoan. This case highlights the importance of considering emerging parasitic agents in cases of hematuria and complex urinary tract infections and underscores the role of molecular diagnostics in identifying atypical and rare pathogens. DESIGN: This is a case report describing the clinical presentation, laboratory findings, and molecular identification of an unusual kinetoplastid organism in a patient's urine. The case is contextualized within the broader and expanding clinical spectrum of human trypanosomatid infections, emphasizing the significance of molecular techniques in detecting emerging and potentially pathogenic organisms. METHODS/RESULTS: Urine microscopy revealed the presence of a motile flagellated protozoan, prompting further investigation. Molecular identification using PCR and sequencing confirmed the organism as Dimastigella trypaniformis, a free-living kinetoplastid from the Rhynchomonadidae family. Previously, D. trypaniformis had only been reported in soil samples from Scotland and termite gut contents in Australia and Germany, with no known associations with vertebrate hosts. This case represents the first potential documented instance of D. trypaniformis in human urine. CONCLUSIONS: The identification of D. trypaniformis in a clinical specimen expands the spectrum of potential urinary pathogens and raises questions about its clinical significance and pathogenic potential. This report underscores the need for heightened awareness of rare and emerging parasitic infections, particularly in patients with atypical presentations. It also highlights the crucial role of molecular diagnostics in identifying novel organisms and guiding appropriate clinical management.

Cadmium (Cd) is a highly toxic heavy metal contaminant found in soil and water due to human activities such as mining and industrial discharge. Cd can accumulate in the body, leading to various health risks such as organ injuries, osteoporosis, renal dysfunction, Type 2 diabetes (T2DM), reproductive diseases, hypertension, cardiovascular diseases, and cancers. The gut is particularly sensitive to Cd toxicity as it acts as the primary barrier against orally ingested Cd. Even at low concentrations, Cd can cause oxidative stress, inflammation, and intestinal bleeding. Cd also disrupts the gut microbiota, affecting its structure, taxonomic composition, and metabolic functions. Cd exposure alters the structure of the gut microbial community, reducing diversity and upregulating certain phyla and genera. This disturbance can lead to physiological and metabolic imbalances, including disruptions in energy homeostasis, amino acid, lipid, nucleotide, and short-chain fatty acid (SCFAs) metabolism. The effects of Cd on the gut microbiota depend on the duration of exposure, the dose of Cd, and can vary based on sex and age. Cd-induced gut dysbiosis has been linked to various diseases, including diabetes, adiposity, atherosclerosis, liver damage, infections, cancer, and neurodegenerative diseases. Interventions targeting the gut microbiota, such as probiotics, specific diets, melatonin, selenium, vitamin D3, and certain compounds, have shown potential in reducing the health risks associated with Cd exposure. However, combined exposure to Cd and other toxicants, such as microplastics (MPs), heavy metals, and antibiotics, can amplify the toxicity and dysbiosis in the gut microbiota.

Polyunsaturated fatty acids (PUFAs) play a crucial role in aiding bacteria to adapt to extreme and stressful environments. While there is a well-established understanding of their production, accrual, and transfer within marine ecosystems, knowledge about terrestrial environments remains limited. Investigation of the intestinal microbiome of earthworms has illuminated the presence of PUFAs presumably of microbial origin, which contrasts with the surrounding soil. To comprehensively study this phenomenon, a multi-faceted approach was employed, combining fatty acid analysis with amplicon sequencing of the PfaA-KS domain of the anaerobic fatty acid synthase gene (pfa), as well as the 16S rRNA and 18S rRNA genes. This methodology was applied to scrutinize the gut microbiome of Eisenia fetida, its compost-based dietary source, and the resultant castings. This study unveiled a distinct gut soil ecosystem from input compost and output castings in fatty acid profile as well as type and abundance of organisms. 16S sequencing provided insights into the microbial composition, showing increased relative abundance of certain Pseudomonadota, including Shewanellaceae, and Planctomycetota, including Gemmataceae within the gut microbiome compared to input bulk soil compost, while Actinomycetota and Bacillota were relatively enriched compared to the casted feces. Sequencing of the PfaA-KS domain revealed amplicon sequence variants (ASVs) belonging primarily to Shewanella. Intriguingly, the 20C PUFAs were identified only in gut soil samples, though PfaA-KS sequence abundance was highest in output castings, indicating a unique metabolism occurring only in the gut. Overall, the results indicate that Shewanella can explain PUFA enrichment in the gut environment because of the pfa gene presence detected via PfaA-KS sequence data.IMPORTANCEPrior research has demonstrated that earthworm microbiomes can potentially harbor polyunsaturated fatty acids (PUFAs) that are not found within their residing soil environment. Moreover, distinct indicator species have been pinpointed for various microbial genera in earthworm microbiomes. Nevertheless, none of these studies have integrated metataxono

Bacterial communities are ubiquitous, found in natural ecosystems, such as soil, and within living organisms, like the human microbiome. The dynamics of these communities in diverse environments depend on factors such as spatial features of the microbial niche, biochemical kinetics, and interactions among bacteria. Moreover, in many systems, bacterial communities are influenced by multiple physical mechanisms, such as mass transport and detachment forces. One example is gut mucosal communities, where dense, closely packed communities develop under the concurrent influence of nutrient transport from the lumen and fluid-mediated detachment of bacteria. In this study, we model a mucosal niche through a coupled agent-based and finite-volume modeling approach. This methodology enables us to model bacterial interactions affected by nutrient release from various sources while adjusting individual bacterial kinetics. We explored how the dispersion and abundance of bacteria are influenced by biochemical kinetics in different types of metabolic interactions, with a particular focus on the trade-off between growth rate and yield. Our findings demonstrate that in competitive scenarios, higher growth rates result in a larger share of the niche space. In contrast, growth yield plays a critical role in neutralism, commensalism, and mutualism interactions. When bacteria are introduced sequentially, they cause distinct spatiotemporal effects, such as deeper niche colonization in commensalism and mutualism scenarios driven by species intermixing effects, which are enhanced by high growth yields. Moreover, sub-ecosystem interactions dictate the dynamics of three-species communities, sometimes yielding unexpected outcomes. Competitive, fast-growing bacteria demonstrate robust colonization abilities, yet they face challenges in displacing established mutualistic systems. Bacteria that develop a cooperative relationship with existing species typically obtain niche residence, regardless of their growth rates, although higher growth yields significantly enhance their abundance. Our results underscore the importance of bacterial niche dynamics in shaping community properties and success

The estimation of postmortem interval (PMI) has long been a focal point in the field of forensic science. Following the death of an organism, microorganisms exhibit a clock-like proliferation pattern during the course of cadaver decomposition, forming the foundation for utilizing microbiology in PMI estimation. The establishment of PMI estimation models based on datasets from different seasons is of great practical significance. In this experiment, we conducted microbiota sequencing and analysis on gravesoil and mouse intestinal contents collected during both the winter and summer seasons and constructed a PMI estimation model using the Random Forest algorithm. The results showed that the MAE of the gut microbiota model in summer was 0.47 ± 0.26 d, R2 = 0.991, and the MAE of the gravesoil model in winter was 1.04 ± 0.22 d, R2 = 0.998. We propose that, in practical applications, it is advantageous to selectively build PMI estimation models based on seasonal variations. Additionally, through a combination of morphological observations, gravesoil microbiota sequencing results, and soil physicochemical data, we identified the time of cadaveric rupture for mouse cadavers, occurring at around days 24-27 in winter and days 6-9 in summer. This study not only confirms previous research findings but also introduces novel insights, contributing to the foundational knowledge necessary to advance the utilization of microbiota for PMI estimation. ©2024 Zhao et al.

The abuse of chemical insecticides has led to strong resistance in cockroaches, and biopesticides with active ingredients based on insect pathogens have good development prospects; however, their slow effect has limited their practical application, and improving their effectiveness has become an urgent problem. In this study, the interaction between Serratia marcescens and Metarhizium anisopliae enhanced their virulence against Blattella germanica and exhibited a synergistic effect. The combination of S. marcescens and M. anisopliae caused more severe tissue damage and accelerated the proliferation of the insect pathogen. The results of high-throughput sequencing demonstrated that the gut microbiota was dysbiotic, the abundance of the opportunistic pathogen Weissella cibaria increased, and entry into the hemocoel accelerated the death of the German cockroaches. In addition, the combination of these two agents strongly downregulated the expression of Imd and Akirin in the IMD pathway and ultimately inhibited the expression of antimicrobial peptides (AMPs). S. marcescens released prodigiosin to disrupted the gut homeostasis and structure, M. anisopliae released destruxin to damaged crucial organs, opportunistic pathogen Weissella cibaria overproliferated, broke the gut epithelium and entered the hemocoel, leading to the death of pests. These findings will allow us to optimize the use of insect pathogens for the management of pests and produce more effective biopesticides.

The transfer of antibiotics and antibiotic resistance (AR) to the soil systems poses ecological hazards to various organisms, including earthworms. Understanding the complex interactions between earthworms, antibiotics, and AR in the soil system requires a comprehensive assessment. Hence, the present review investigates the behaviour, fate, impacts, and mechanisms involved in the interaction of earthworms with antibiotics and AR. The antibiotics and AR detected in earthworms and their associated media, such as vermicompost, are presented, but several other antibiotics and AR widely detected in soils remain understudied. As receptors and bioassay organisms, earthworms are adversely affected by antibiotics and AR causing (1) acute and chronic toxicity, and (2) emergence of AR in previously susceptible earthworm gut microbiota, respectively. The paper also highlights that, apart from this toxicity, earthworms can also mitigate against antibiotics, antibiotic-resistant bacteria and antibiotic-resistance genes by reducing bacterial diversity and abundance. The behaviour and fate processes, including biodegradation pathways, biomarkers of antibiotics and AR in earthworms, are discussed. In addition, the factors controlling the behaviour and fate of antibiotics and AR and their interactions with earthworms are discussed. Overall, earthworms mitigate antibiotics and AR via various proximal and distal mechanisms, while dual but contradictory functions (i.e., mitigatory and facilitatory) were reported for AR. We recommend that future research based on the One-World-One-Health approach should address the following gaps: (1) under-studied antibiotics and AR, (2) degradation mechanisms and pathways of antibiotics, (3) effects of environmentally relevant mixtures of antibiotics, (4) bio-augmentation in earthworm-based bioremediation of antibiotics, (5) long-term fate of antibiotics and their metabolites, (6) bio-transfers of antibiotics and AR by earthworms, (7) development of earthworm biomarkers for antibiotics and AR, (8) application of earthworm-based bioremediation of antibiotics and AR, (9) cascading ecological impacts of antibiotics and AR on earthworms, and (10) pilot

BACKGROUND: A low-clean living environment (LCLE) can increase gut microbial diversity and prevent allergic diseases, whereas gut microbial dysbiosis is closely related to the pathogenesis of asthma. Our previous studies suggested that soil in the LCLE is a key factor in shaping intestinal microbiota. We aimed to explore whether sterilized soil intake as a prebiotic while being incubated with microbes in the air can attenuate mouse asthma inflammation by modifying gut microbiota. METHODS: 16S rRNA gene sequencing was used to analyze the gut microbial composition, in combination with immune parameters measured in the lung and serum samples. RESULTS: 16S rRNA gene sequencing results showed significant differences in the fecal microbiota composition between the test and control mice, with a higher abundance of Allobaculum, Alistipes, and Lachnospiraceae_UCG-001, which produce short-chain fatty acids and are beneficial for health in the test mice. Soil intake significantly downregulated the concentrations of IL-4 and IL-9 in serum and increased the expression of IFN-γ, which regulated the Th1/Th2 balance in the lung by polarizing the immune system toward Th1, alleviating ovalbumin-induced asthma inflammation. The effect of sensitization on gut microbiota was greater than that of air microbes and age together but weaker than that of soil. CONCLUSIONS: Soil intake effectively reduced the expression of inflammatory cytokines in asthmatic mice, possibly by promoting the growth of multiple beneficial bacteria. The results indicated that the development of soil-based prebiotic products might be used for allergic asthma management, and our study provides further evidence for the hygiene hypothesis. © 2024 The Authors.