Search Results (1,169)

The human microbiome influences health and disease through diverse biochemical and functional outputs (e.g., enzymes, structural proteins, metabolites, and other cellular components) that affect nearly every aspect of human physiology. Metatranscriptomics (MT), an unbiased RNA sequencing approach, is a high-throughput and high-content method that quantifies both gut microbial taxonomy and active biochemical functions. Because microbial community composition and gene expression are dynamic, understanding temporal variation in the gut metatranscriptome across multiple time scales is essential. Here, we report the temporal dynamics of gut microbiome species and functions using a large cohort (n = 6157) with a clinically validated stool MT test. We quantified microbiome stability from hours to years and assessed taxonomic and functional resilience to major luminal perturbations, such as colonoscopy bowel preparation. Longitudinal analyses of samples collected within the same day, and across days, weeks, months, and years, revealed consistently high stability in both composition and gene expression within a single day and, importantly, across an approximate six-month period. Among individuals reporting stable diets and no antibiotic exposure, taxonomic and functional profiles remained stable for up to three years. Following colonoscopy preparation, our preliminary study of the microbiome demonstrated strong resilience, returning to its pre-procedure state within one week. Overall, these findings demonstrate that the gut microbiome is generally stable over a six-month time frame, with longer-term changes occurring gradually. These findings support the robustness of stool-based MT profiling for species-level and pathway-resolved functional analysis in longitudinal research and health applications. Full article

Salmonella is a major cause of foodborne illness worldwide, posing significant risks to public health and food safety. This study investigated the prevalence, serovar distribution, genotypic characteristics, and antimicrobial resistance (AMR) profiles of Salmonella. A total of 2515 food samples were collected from retail markets, supermarkets, and food processing facilities, and 13,670 stool samples were obtained from sentinel hospitals across 14 cities in Liaoning. The Kruskal–Wallis test was used to compare genetic features among serovars, followed by Dunn’s post hoc test for pairwise comparisons. A total of 314 Salmonella strains were identified, with raw poultry showing the highest detection rate (28.88%) among food sources and children aged 0–6 years (3.47%) the highest among the clinical age groups. Among food samples, S. Enteritidis was the most prevalent serovar (42.6%), and it was also the most common in clinical samples (35.8%); in contrast, S. 4,[5],12:i:- was dominant in pediatric clinical cases. According to AMR analysis, 90.13% of strains were resistant to at least one antibiotic and 67.83% were multidrug-resistant (MDR), with the highest resistance to ampicillin (68.47%). Analysis revealed that S. 4,[5],12:i:- harbored the ASSuT resistance module (blaTEM-1B, aph(3″)-Ib/aph(6)-Id, sul2, tet(B)). Extensive MDR phenotypes were observed in S. Indiana and S. Kentucky, associated with abundant insertion sequences (IS) and resistance genes (ARGs), including clinically critical determinants (blaNDM-9, mcr-1.1, rmtB). The highest mean virulence factor (VF) count (111.17) was observed in S. Enteritidis, contributing to its epidemiological success. Conversely, S. Indiana and S. Kentucky, predominantly food-associated serovars, exhibited reduced virulence but served as critical AMR reservoirs. These findings highlight the epidemiological characteristics and AMR risks of Salmonella in food and clinical settings, providing critical data for food safety and clinical antimicrobial stewardship. Full article

This study aimed to explore the efficacy of oral supplementation with 11 probiotic strains, combined in the strain mix Probatech™ (Centro Sperimentale del Latte S.r.l Strada Provinciale per Merlino, 326839 Zelo Buon Persico (LO), Italy) and delivered through the food supplement PROBAFLOR and how it plays a positive role in maintaining normal intestinal function, providing benefits in healthy adult subjects. A 16-week, randomized, double-blind, placebo-controlled clinical study was conducted starting with 60 participants. Participants were randomly assigned to either the probiotic or placebo group. Participants were asked to provide one faecal sample at the beginning of the study, another one after 12 weeks of supplementation and the final one after 16 weeks. Amplicon 16S rRNA gene sequencing and GC-MS Short-Chain Fatty Acid (SCFA) profiling were performed on the faecal samples. Participants filled out questionnaires to assess their gastrointestinal health and psychological well-being. The overall mean GIQLI scores increased in both groups over time. The increases were significant within both groups but not between groups. Following the administration of PROBAFLOR, Shannon and Simpson diversity indices showed a significant increase at day 90 (week 12) (p < 0.05), demonstrating that the intervention effectively enhanced gut microbiota diversity. A shift in the intestinal microbiota towards SCFA-producing families and genera was observed. Moreover, the change in total and single SCFAs was significantly different between probiotic and placebo groups at the end of the supplementation period. Once-daily consumption of the PROBAFLOR probiotics formula regulated gut microbiota balance by modulating SCFA production. It may be beneficial for gut health, improving defecation habits and satisfaction, normalizing stool frequency, and promoting bacterial metabolism. Full article

Background: Functional constipation (FC) in children is characterized by infrequent bowel movements and straining or painful defecation, often accompanied by abdominal pain and fecal incontinence. Typically, in cases without organic pathology, it is diagnosed as this disease. When diagnosed according to Rome IV criteria, the global prevalence of this condition is 14.4%. Additionally, one study reported that in the UK, one in seven adults and one in three children suffer from constipation. In just one year (2018–2019), the National Health Service incurred £168 million in costs related to constipation, with over 175,000 hospital days consumed and numbers continuing to rise. Massage therapy is viewed as a promising treatment modality, though systematic evaluation evidence for its use in combination therapies remains insufficient. Research Objective: This study aims to systematically evaluate the efficacy and safety of tuina massage as an adjunct to conventional treatment for pediatric functional constipation through meta-analysis. Materials and Methods: Eight databases were searched to collect relevant randomized controlled trials (RCTs) from their inception to 23 January 2026. Primary outcomes included clinical total effective rate and BSFS score. Secondary outcomes comprised adverse reaction incidence, recurrence rate, defecation difficulty, and improvement in bowel movement frequency. Quality assessment was performed using the Cochrane Risk of Bias tool (RoB 2). Meta-analysis was conducted using RevMan 5.4 and Stata 13 software. Results: A total of 16 RCTs involving 1387 pediatric patients were included. Meta-analysis revealed that compared with conventional therapy alone, the combination of Tuina and conventional therapy significantly improved the overall clinical response rate (RR = 1.18, 95% CI 1.10 to 1.25, p < 0.00001; Tau² = 0.01), improved the Bristol Stool Form Scale (BSFS) score (MD = 0.55, 95% CI 0.32 to 0.78, p < 0.00001; Tau² = 0.05), and reduced the defecation difficulty score (MD = −1.36, 95% CI −1.75 to −0.98, p < 0.00001; Tau² = 0.18). However, substantial heterogeneity was observed across these outcomes (I² = 75%, 71%, and 96%, respectively). The 95% prediction intervals crossed the null value for all three primary outcomes (treatment success rate: 0.95–1.47; BSFS score: −0.25 to 1.35; defecation difficulty score: –2.85 to 0.13), indicating that the true effect may vary substantially across future study settings. Regarding recurrence, Tuina-assisted therapy resulted in a lower recurrence rate compared to conventional therapy alone (RR = 0.27, 95% CI 0.16 to 0.47, p < 0.00001). While improvements in weekly bowel movement frequency were reported, they could not be confirmed due to insufficient sample size. It remains unclear whether Tuina can mitigate adverse effects associated with control group treatments. Conclusions: Current evidence suggests that tuina as an adjunct to conventional treatment may offer improvements in treatment success, stool consistency, defecation difficulty, and recurrence rates in children with functional constipation. However, given the substantial heterogeneity (I² up to 96%), wide prediction intervals that crossed the null value for all three primary outcomes, methodological limitations of the included studies (e.g., lack of blinding, unclear randomization), and short follow-up periods (most ≤2 months), these findings should be interpreted as exploratory rather than definitive. Evidence on weekly stool frequency and adverse reactions remains inconclusive. High-quality, long-term trials with standardized outcome measures and rigorous blinding are needed to validate these preliminary findings. Full article

Background/Objectives: Mental health conditions, including stress, anxiety, depression, and sleep problems, represent a significant health concern globally. Mounting evidence suggests a link between mental health and the gut microbiome via the gut–brain axis. However, discrepancies in human microbiome data exist due to the heterogeneity in study design and analytical approaches. Thus, this study aimed to explore the gut microbial characteristics associated with self-reported mental health symptoms using multiple analytical methods. Methods: A total of 44 healthy adults, defined as individuals without any major chronic medical conditions, were assessed for mental health symptoms using self-reported questionnaire data. To evaluate gut microbial characteristics, stool samples were collected at six time points over 28 days and underwent 16S rRNA gene sequencing. Differential abundance was assessed via ANCOM-BC, and a random forest classifier was implemented to rank features important for the classification of mental health symptoms. Participants who did not report anxiety, stress, depression, or sleep problems served as the reference group for microbiome comparisons. Results: The proportion of participants with self-reported mental health symptoms was 11.4% (stress), 27.3% (depression), 31.8% (anxiety), and 15.9% (sleep problems). Participants reporting mental health symptoms showed differences in gut microbiome composition compared to asymptomatic participants, including variation in alpha- and beta-diversity. Differential analysis identified specific taxa with higher or lower relative abundance in participants reporting specific mental health symptoms. Random forest feature ranking identified partially overlapping taxa across methods, suggesting candidate associations warranting further investigation. Conclusions: These exploratory findings suggest that self-reported mental health symptoms in otherwise healthy adults are associated with differences in the gut microbiome. The taxa identified in this study represent candidates for validation in larger, independent cohorts. Full article

Background and Aims: Severe alcohol-associated hepatitis (SAH) can trigger unstable decompensations in cirrhosis patients. They experience high rates of emergency department visits and hospitalization. We evaluated real-world clinical outcomes following palliative-faecal microbiota transplantation (pFMT) compared to best supportive care (BSC) in this critically ill population. Patients and Methods: From July 2021 to April 2024, 28 patients on pFMT were compared with 37 on BSC. Patients on pFMT received nasoduodenal healthy donor stool infusion daily for 5-days. Patients were followed up for portal hypertension-related events, infections, hospitalizations, extrahepatic organ failure and 6- and 12-months survival. 16S rRNA sequencing on stool samples collected at baseline and on follow up were analysed for changes in relative abundance (RA) of bacterial communities. Results: Patients were matched for age, type of decompensation and liver disease severity at enrolment. Twelve-month survival was 64.3% in pFMT versus 51.4% in BSC groups. pFMT dramatically reduced hospital readmissions (mean 0.76 ± 0.76 vs. 2.29 ± 1.27, p < 0.001). Unstable decompensations beyond 3 months occurred in 14.3% of pFMT versus 64.9% of BSC (p < 0.001). Organ failures were lesser with pFMT: acute kidney injury 7.7% versus 93.8% (p < 0.001), hepatic encephalopathy 7.1% versus 68.2% (p < 0.001). Infection burden was significantly lower (53.6% vs. 83.8%, p = 0.008), particularly infections requiring admission (17.4% vs. 66.7%, p < 0.001) with pFMT. Microbiome analysis revealed progressive expansion of Gram-negative genera in BSC, and beneficial Actinobacteria in pFMT-treated patients at 3, 6, and 12 months. Conclusions: Palliative FMT represents a unique disease-modifying intervention in end-stage alcohol-related cirrhosis, preventing organ failure progression, reducing healthcare utilization, and improving survival trajectories. Full article

Background: The Firmicutes/Bacteroidetes (F/B) ratio has been proposed as a microbial biomarker of obesity and metabolic alterations; however, its reliability remains controversial, particularly in young populations. Methods: This study evaluated the relationship between the F/B ratio, body fat percentage, and metabolic markers in 70 university students aged 18–25 years, classified as normal weight (29.5%), overweight (27.4%), or obese (43.2%). Anthropometric measurements and biochemical parameters (glucose, triglycerides, total cholesterol, and HDL-C) were obtained using standard methods, and stool samples were analyzed to determine the F/B ratio. Results: Mean glucose and cholesterol were within normal ranges, whereas triglycerides showed high variability, and HDL-C was lower in men. Although the F/B ratio increased across nutritional groups, regression analyses showed weak correlations (R < 0.5) and no significant associations (p > 0.05). Conclusions: The F/B ratio is not an adequate standalone indicator of metabolic status in Mexican young adults. Full article

Background: Schistosoma spp. are trematodes occurring in tropical endemic areas but can be imported to non-endemic regions as causes of travel-associated infections. In this study, two pan-trematode-specific real-time PCR assays were evaluated for their diagnostic sensitivity in detecting Schistosoma spp. DNA in diagnostic human samples. Methods: Two previously described pan-trematode-specific real-time PCR assays were comparatively assessed using diagnostic samples containing DNA of either the S. haematobium complex or the S. mansoni complex, as confirmed by Schistosoma species complex-specific real-time PCR. Results: Out of a total of 655 samples containing Schistosoma spp. DNA, positive signals in at least one of the two pan-trematode real-time PCR assays were recorded for 17 (2.6%) nucleic acid extractions. Although sensitivity was in the >90% range for stool samples, only a few individual blood plasma and serum samples, and none of the Schistosoma spp. DNA-containing tissue or urine samples, tested positive by pan-trematode PCR. The lower sensitivity of pan-trematode PCR compared with Schistosoma spp.-specific PCR was semi-quantitatively confirmed by higher cycle threshold (Ct) values in the former. When comparing samples with concordant versus discordant positive results for Schistosoma spp.-specific and pan-trematode PCR, Ct values of the Schistosoma spp.-specific PCR were lower in concordantly positive samples than in discordantly positive samples. Conclusions: While the assessed pan-trematode PCR assays showed insufficient sensitivity as screening tools for blood plasma, blood serum, tissue, and urine samples from individuals with suspected schistosomiasis, they were sufficiently sensitive when applied to stool samples, in which substantial amounts of target DNA, as indicated by low Ct values in the Schistosoma species complex-specific real-time PCR assays, can be expected. For screening for Schistosoma spp. DNA in sample materials other than stool, the use of highly sensitive target-specific PCR remains necessary. Full article

Background: This study evaluated the inter-method agreement of an in-house qualitative CMV real-time PCR assay for the detection of cytomegalovirus (CMV) DNA in various non-plasma clinical specimen types, in comparison with a commercially available comparator assay. Methods: In this prospective comparative study, 186 clinical specimens—including bronchoalveolar lavage fluid (BALF), stool, urine, colonoscopic biopsy, amniotic fluid, and intraocular fluid—were analyzed. A total of 166 samples with valid results from both test systems were included in the inter-method comparison. CMV DNA was detected using the in-house qualitative PCR assay in parallel with the comparator assay (artus^® CMV QS-RGQ kit). Agreement was assessed using positive percent agreement (PPA), negative percent agreement (NPA), overall percent agreement (OPA), and Cohen’s kappa coefficient (κ), in accordance with CLSI EP12-A2 recommendations. Results: Substantial overall inter-method agreement was observed when all specimens were evaluated collectively (κ = 0.66). Agreement metrics were highest in stool, urine, and invasive specimens, whereas BALF samples demonstrated comparatively lower agreement, reflecting potential matrix-related analytical variability. Conclusion: The laboratory-developed qualitative CMV PCR assay demonstrated substantial inter-method agreement with the comparator assay across multiple non-plasma specimen types. The findings highlight specimen-specific variability in qualitative CMV DNA detection and represent analytical concordance between two molecular assays rather than definitive clinical diagnostic accuracy or viral load quantification. Full article

Objectives: We aimed to compare the treatment success and patient compliance of triple therapy with levofloxacin, high-dose dual therapy with amoxicillin, and quadruple therapy with bismuth and levofloxacin as first-line treatment for Helicobacter pylori (H. pylori) eradication using amoxicillin 875 mg + clavulanic acid 125 mg instead of amoxicillin 1 g. Methods: Patients who tested positive for Helicobacter pylori in the histopathological examination of biopsies taken during upper gastrointestinal endoscopy were initially divided into three different treatment groups. There were 179 patients in the first group, 178 patients in the second group, and 182 patients in the third group. A total of 480 patients from these groups who came for follow-up were included in this study, with 160 patients in each group receiving one of three different treatment protocols. The first group received treatment with amoxicillin 875 mg + 125 mg clavulanic acid twice daily, levofloxacin 500 mg once daily, and pantoprazole 40 mg twice daily (Group 1). The second group received treatment with amoxicillin 875 mg + 125 mg clavulanic acid three times a day, along with pantoprazole 40 mg twice daily (Group 2) as treatment. The third group received treatment with amoxicillin 875 mg + 125 mg clavulanic acid, twice daily, levofloxacin 500 mg once daily, pantoprazole 40 mg twice daily, and bismuth subsalicylate 262 mg2 pieces four times a day (Group 3). H. pylori was checked with a stool antigen test 45 days after the 14-day treatment. The groups were compared in terms of treatment success and treatment compliance. Results: In Group 1, 150 (90.6%) of 160 patients tested negative for an H. pylori antigen in stool samples on day 45 after treatment. This rate was 139 (86.9%) in Group 2 and 148 (92.5%) in Group 3. There were no statistically significant differences between the three groups in terms of treatment success (p = 0.233). Side effects were observed in 10 (6.2%) patients in Group 1. Side effects were present in nine (5.6%) patients in Group 2. Side effects were observed in 12 (7.5%) patients in Group 3. There was no significant difference between the groups in terms of patient compliance (p = 0.786). Conclusions: Treatment success and side effects were similar in all three groups, with no statistical difference. The combination of amoxicillin 875 mg + clavulanic acid 125 mg is at least as effective as amoxicillin 1 g alone. Full article

Background: Fecal calprotectin (FC) is a well-established, non-invasive biomarker of intestinal inflammation and is widely used to differentiate inflammatory bowel disease (IBD) from functional gastrointestinal disorders. Although enzyme-linked immunosorbent assays (ELISA) remain the reference method, rapid immunochromatographic tests (ICTs) offer important operational advantages for point-of-care (POC) diagnostics. However, variability in analytical performance among available ICTs remains a concern. Objective: This study aimed to evaluate the diagnostic accuracy of the CerTest Calprotectin one-step card (CerTest Biotec S.L., Zaragoza, Spain) in comparison with the Actim^® Calprotectin lateral flow assay and the reference Calprest^® ELISA (Eurospital Diagnostics, Italy). Methods: A total of 128 fresh stool samples from patients clinically suspected of IBD were analyzed in parallel using all three assays. For the reference ELISA (Calprest^®), a cutoff value of >40 µg/g was applied according to the manufacturer’s instructions. For discrepant results between assays, a cutoff of 200 ng/mL (equivalent to 200 µg hCp/g stool) was employed for ELISA Calprest^® to resolve inconsistencies. The results of the lateral flow assays (CerTest^® Calprotectin ICT and Actim^® Calprotectin) were interpreted using their respective manufacturer-recommended thresholds. Diagnostic sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated using ELISA as the reference standard. Agreement between methods was assessed using Cohen’s κ coefficient. Results: Using ELISA, 47 of 128 samples (36.7%) exceeded the 40 µg/g cutoff. Compared with the Actim^® assay, the CerTest card demonstrated a sensitivity of 88.0% (95% CI: 75.7–95.5), a specificity of 100.0% (95% CI: 95.4–100), and a strong agreement (κ = 0.90). When compared with ELISA, the CerTest assay showed a sensitivity of 87.2% (95% CI: 74.3–95.2), a specificity of 96.3% (95% CI: 89.6–99.2), a PPV of 93.2%, an NPV of 93.2%, and a strong agreement (κ = 0.85). Conclusions: The CerTest Calprotectin one-step card provides a rapid and reliable detection of fecal calprotectin, demonstrating a high sensitivity and specificity that are comparable to both other lateral flow assays and the ELISA reference method. These findings support the use of rapid immunochromatographic testing as a valuable tool for preliminary screening and clinical decision-making in patients suspected of IBD, while acknowledging that histology remains the gold standard for definitive diagnosis. Full article

Background: In this study, we aimed to compare metabolomic profiles, biodistribution, and detoxification patterns of the novel SN-38 derivative NMe with irinotecan (IR), and to identify NMe-specific metabolites to evaluate its preclinical pharmacokinetic advantages. Methods: In vivo ADME studies were conducted for NMe, a 9-aminomethyl SN-38 derivative, and IR following a single intraperitoneal dose of 40 mg/kg in mice. Additionally, ADMET properties were predicted using ADMETlab and SwissADME tools for comparison. Levels of NMe and irinotecan absorbed into plasma, distributed to tissues, and metabolized were monitored in liver, lung, spleen, kidney, and stool samples at 15, 30, and 60 min post-administration. Tissue extracts were analysed using high-performance liquid chromatography (HPLC), liquid chromatography–electrospray ionization quadrupole time-of-flight-tandem mass spectrometry (LC-ESI-QTOF-MS), and nuclear magnetic resonance (NMR) techniques after lyophilization and reconstitution. We compared the metabolomic profiles of irinotecan and NMe. Results: We identified and confirmed NMe-specific metabolites, including 9-CH₂-S-cysteine conjugate, 9-CH₂OH, and NMe-formyl. Notably, novel irinotecan metabolites (IR-OH and IR-ΔE) were detected in small amounts in kidney samples. In some cases, two literature-known photodegradation products of irinotecan were present. NMe was found to quickly metabolize with different distribution to tissues, significantly greater to kidney and liver. Two SN-38 glucuronides, SN-38G(α) and SN-38G(β), were detected corresponding to α- and β-anomers. Where it was possible, NMe, IR and SN-38 were quantified using external calibration curves. In IR group, controlled and prolonged release of SN-38 was confirmed in all samples, yet SN-38G was observed in minority only in plasma, kidney, or lungs. In NMe groups, great relative amounts of SN-38 and SN-38G were detected. Greater content of SN-38G in NMe group than in irinotecan is expected to contribute to modulation and alleviation of some side effects in irinotecan-involved therapies, such as gastrointestinal toxicities (GIT). Conclusions: NMe shows a distinct metabolic profile characterized by rapid biotransformation, higher systemic glucuronidation of SN-38, and formation of unique metabolites, suggesting a potentially wider therapeutic window and reduced toxicity compared with IR. Full article

Background: Monitoring inflammatory bowel disease (IBD) during pregnancy is challenging due tolimited use of invasive tools. While fecal calprotectin is considered reliable, its use is limited by patient adherence and availability. Blood-based markers, such as serum chitinase-3-like-1 (CHI3L1), offer a promising alternative. Aim: To evaluate whether serum CHI3L1 reflects disease activity in pregnant IBD patients, compared to standard markers and clinical questionnaires. Methods: Pregnant IBD patients were recruited from a multidisciplinary clinic. Blood samples were collected to assess inflammatory markers. Stool samples were used to measure calprotectin levels. Each visit was classified as a distinct sample for analysis. Correlations between CHI3L1 and disease activity markers were examined. Results: A total of 124 samples from 80 pregnant IBD patients were analyzed: 90 from Crohn’s disease (CD) patients and 34 from ulcerative colitis (UC) patients. CHI3L1 levels showed a significant positive correlation with fecal calprotectin (r_p = 0.366, p = 0.008), ESR (r_p = 0.358, p = 0.001), CRP (r_p = 0.478, p < 0.001) and standardized clinical scoring questionnaires. Elevated CHI3L1 (>56.6 ng/mL) is a risk factor for active disease (OR 8.78, 95% CI 1.54–49.83, p = 0.014). Conclusions: Serum CHI3L1 is positively associated with established markers of inflammation and may serve as a useful non-invasive biomarker for monitoring IBD activity during pregnancy, a medical condition in which invasive procedures are not recommended. Based on CHI3L1 levels, personalized treatment for pregnant IBD patients can be tailored. However, further validation is recommended. Full article

Cancer is a major health concern, with its incidence rate continuing to increase. There is growing interest in the microbiota and its role in carcinogenesis, as it significantly influences physiological and pathological processes. Various aspects of the microbiome have been shown to have both anti-tumor and pro-tumor effects. Advances in techniques such as high-throughput DNA sequencing have greatly improved our understanding of microbial populations in the human and canine gut. We aimed to (1) characterize the intestinal microbiota of healthy dogs and dogs with cutaneous mast cell tumors (MCTs), (2) assess changes in the intestinal microbiota of dogs undergoing electrochemotherapy (ECT) combined with gene electrotransfer (GET) of the IL-12 plasmid (IL-12), and (3) explore possible associations with the expression of immune markers Programmed cell death protein 1 (PD-1), Programmed death-ligand 1 (PD-L1), and Granzyme B (GZMB) in MCT tissue. Stool samples were collected from healthy dogs (n = 24) and dogs with MCTs (n = 24) before and after ECT and IL-12 GET. DNA was extracted from the samples, and shallow shotgun sequencing was performed. Immunohistochemistry was performed on the tumors to assess the expression of PD-1, PD-L1, and GZMB. The dysbiosis index, alpha diversity, and beta diversity did not differ between groups. Regarding microbial composition, Bifidobacterium animalis, Corynebacterium variabile, Lactobacillus johnsonii, Pediococcus pentosaceus, Streptococcus anginosus, Streptococcus equinus, Streptococcus intermedius, Clostridium thermobutyricum, Megasphaera elsdenii, and Anaerobiospirillum sp. were found in lower relative abundance in feces of dogs with MCTs, while Bacteroides togonis, Lactobacillus amylolyticus, Prevotella sp. CAG:279, and Megamonas hypermegale were more abundant compared to healthy dogs. Our study provides further insight into the composition of the gut microbiota in dogs with MCTs, where ECT and IL-12 GET did not lead to major shifts. We were unable to establish any association between the expression of immune markers and the microbiota. Full article

Colorectal cancer (CRC) continues to represent a substantial worldwide health burden. Accurate risk classification and early detection have a significant impact on prognosis. There is still a significant percentage of patients who are diagnosed at advanced stages, notwithstanding the progress that has been made in screening and treatment. Thus, improved molecular tools that encompass the biological complexity of CRC are needed. High-throughput technologies have expanded the biomarker array for CRC screening, prognosis, and therapeutic prediction. This review summarizes evidence on established and emerging molecular tools from tumor tissue, blood, and stool samples, such as DNA mutations, methylation markers, RNA signatures, circulating tumor DNA (ctDNA), circulating cell-free DNA (cfDNA), extracellular vesicles, and multi-omic composite assays. These provide alternatives to conventional approaches that are relatively less invasive and more sensitive. Prognostic biomarkers—such as RAS, BRAF, HER2 alterations, mismatch repair deficiency, tumor mutational burden, methylation signatures, and non-coding RNAs—provide insight into tumor behavior and recurrence risk. To guide targeted therapies, immunotherapies, and chemotherapy response, predictive biomarkers such as RAS/BRAF mutations, HER2 amplification, MSI-H/dMMR status, POLE/POLD1 mutations, DNA methylation panels, miRNAs, lncRNAs, and liquid biopsy markers are crucial. Emerging technologies such as multi-omics, AI-enhanced biomarker discovery, and novel liquid biopsy components (evDNA, circRNAs) pave the way to precision oncology. These molecular tools have the potential to change how CRC is managed by earlier detection and more precise predictive biomarkers. However, large-scale validation and clinical standardization are still crucial for their extensive utilization. Full article