Comparative analysis of gut microbiota composition in the fecal samples from type 2 diabetes mellitus patients and healthy individuals: a case control study
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Abstract
Background and Objectives: Insulin resistance and elevated blood glucose levels are the hallmarks of Type 2 Diabetes Mellitus (T2DM), a chronic metabolic condition. Emerging research suggests that gut microbiota may play a causal role in T2DM. This study compares T2DM patients' gut microbiota to healthy controls, focusing on Lactobacillus, Bifidobacterium, Akkermansia muciniphila, Prevotella, Bacteroidetes, and Firmicutes.
Materials and Methods: This case-control research involved 50 T2DM patients and 50 healthy controls, aged 39-75. Quantitative real-time PCR (qPCR) employing 16S rRNA gene primers was used to detect and quantify bacterial diversity in fecal samples. Statistical analyses were performed to compare the microbiota composition between groups.
Results: The gut microbiome of patients with Type 2 Diabetes Mellitus differed significantly from that of healthy controls. In T2DM patients, Lactobacillus spp. and the Firmicutes phylum had higher relative fold differences, while A. muciniphila had lower abundance. No substantial alterations were seen in Bifidobacterium spp., Prevotella, or Bacteroidetes. T2DM patients had more Lactobacillus spp. and Firmicutes and less A. muciniphila in their gut microbiome.
Conclusion: While gut microbiota is linked to T2DM, this study analyzes the bacterial composition to identify taxa that change significantly. Further research is essential to unravel the complex relationships between gut microbiota and T2DM pathogenesis, particularly through species-level analysis and genomic studies to identify the primary associated clades.
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13. Tanase DM, Gosav EM, Neculae E, Costea CF, Ciocoiu M, Hurjui LL, et al. Role of gut microbiota on onset and progression of microvascular complications of type 2 diabetes (T2DM). Nutrients 2020; 12: 3719.
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15. Zhao S, Xiang J, Abedin M, Wang J, Zhang Z, Zhang Z, et al. Characterization and anti-inflammatory effects of Akkermansia muciniphila-derived extracellular vesicles. Microorganisms 2025; 13: 464.
16. Koval S, Snihurska I. Role of gut microbiota in the pathogenesis of type 2 diabetes mellitus (literature review). Med Perspekt 2021; 26: 22-30.
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18. Carding S, Verbeke K, Vipond DT, Corfe BM, Owen LJ. Dysbiosis of the gut microbiota in disease. Microb Ecol Health Dis 2015; 26: 26191.
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20. Fong W, Li Q, Yu J. Gut microbiota modulation: a novel strategy for prevention and treatment of colorectal cancer. Oncogene 2020; 39: 4925-4943.
21. Sedighi M, Razavi S, Navab-Moghadam F, Khamseh ME, Alaei-Shahmiri F, Mehrtash A, et al. Comparison of gut microbiota in adult patients with type 2 diabetes and healthy individuals. Microb Pathog 2017; 111: 362-369.
22. Candela M, Biagi E, Soverini M, Consolandi C, Quercia S, Severgnini M, et al. Modulation of gut microbiota dysbioses in type 2 diabetic patients by macrobiotic Ma-Pi 2 diet. Br J Nutr 2016; 116: 80-93.
23. Lê KA, Li Y, Xu X, Yang W, Liu T, Zhao X, et al. Alterations in fecal Lactobacillus and Bifidobacterium species in type 2 diabetic patients in Southern China population. Front Physiol 2013; 3: 496.
24. Wu T, Zhang Y, Li W, Zhao Y, Long H, Muhindo EM, et al. Lactobacillus rhamnosus LRa05 ameliorate hyperglycemia through a regulating glucagon-mediated signaling pathway and gut microbiota in type 2 diabetic mice. J Agric Food Chem 2021; 69: 8797-8806.
25. Gulnaz A, Nadeem J, Han J-H, Lew L-C, Son J-D, Park Y-H, et al. Lactobacillus SPS in reducing the risk of diabetes in high-fat diet-induced diabetic mice by modulating the gut microbiome and inhibiting key digestive enzymes associated with diabetes. Biology (Basel) 2021; 10: 348.
26. Remely M, Hippe B, Zanner J, Aumueller E, Brath H, Haslberger AG. Gut microbiota of obese, type 2 diabetic individuals is enriched in Faecalibacterium prausnitzii, Akkermansia muciniphila and Peptostreptococcus anaerobius after weight loss. Endocr Metab Immune Disord Drug Targets 2016; 16: 99-106.
27. Qin J, Li Y, Cai Z, Li S, Zhu J, Zhang F, et al. A metagenome-wide association study of gut microbiota in type 2 diabetes. Nature 2012; 490: 55-60.
28. El-Sabbagh A, El Sayed Zaki M, Motawea MM, Alkasaby NM. Molecular study of lactobacilli species in patients with Type 2 Diabetes Mellitus. Open Microbiol J 2022; 16: 10.2174/18742858-v16-e2205090.
29. Larsen N, Vogensen FK, van den Berg FW, Nielsen DS, Andreasen AS, Pedersen BK, et al. Gut microbiota in human adults with type 2 diabetes differs from non-diabetic adults. PloS One 2010; 5(2): e9085.
30. Tsai HJ, Tsai YC, Hung WW, Hung WC, Chang CC, Dai CY. Gut microbiota and non-alcoholic fatty liver disease severity in type 2 diabetes patients. J Pers Med 2021; 11: 238.
31. Ebrahimzadeh Leylabadlo H, Ghotaslou R, Samadi Kafil H, Feizabadi MM, Moaddab SY, Farajnia S, et al. Non-alcoholic fatty liver diseases: from role of gut microbiota to microbial-based therapies. Eur J Clin Microbiol Infect Dis 2020; 39: 613-627.
32. Murri M, Leiva I, Gomez-Zumaquero JM, Tinahones FJ, Cardona F, Soriguer F, et al. Gut microbiota in children with type 1 diabetes differs from that in healthy children: a case-control study. BMC Med 2013; 11: 46.
33. Zhang J, Ni Y, Qian L, Fang Q, Zheng T, Zhang M, et al. Decreased abundance of Akkermansia muciniphila leads to the impairment of insulin secretion and glucose homeostasis in lean type 2 diabetes. Adv Sci (Weinh) 2021; 8(16): e2100536.
34. Zhang L, Qin Q, Liu M, Zhang X, He F, Wang G. Akkermansia muciniphila can reduce the damage of gluco/lipotoxicity, oxidative stress and inflammation, and normalize intestine microbiota in streptozotocin-induced diabetic rats. Pathog Dis 2018; 76: 10.1093/femspd/fty028.
35. Sanjiwani MID, Aryadi IPH, Semadi IMS. Review of literature on Akkermansia muciniphila and its possible role in the etiopathogenesis and therapy of type 2 diabetes mellitus. J ASEAN Fed Endocr Soc 2022; 37: 69-74.
36. Rodrigues VF, Elias-Oliveira J, Pereira ÍS, Pereira JA, Barbosa SC, Machado MSG, et al. Akkermansia muciniphila and gut immune system: a good friendship that attenuates inflammatory bowel disease, obesity, and diabetes. Front Immunol 2022; 13: 934695.
2. Al-Kuwari SH (2024). Diabetes in Cross-Cultural Perspective, in Arab Americans in the United States: Immigration, Culture and Health. pp. 9-23.
3. DeFronzo RA, Ferrannini E, Groop L, Henry RR, Herman WH, Holst JJ, et al. Type 2 diabetes mellitus. Nat Rev Dis Primers 2015; 1: 15019.
4. Yang X, Sun J, Zhang W. Global trends in burden of type 2 diabetes attributable to physical inactivity across 204 countries and territories, 1990-2019. Front Endocrinol (Lausanne) 2024; 15: 1343002.
5. Pires L, González-Paramás AM, Heleno SA, Calhelha RC. The role of gut microbiota in the etiopathogenesis of multiple chronic diseases. Antibiotics (Basel) 2024; 13: 392.
6. Doroszkiewicz J, Mroczko J, Winkel I, Mroczko B. Metabolic and Immune System Dysregulation: Unraveling the connections between alzheimer’s disease, diabetes, inflammatory Bowel diseases, and Rheumatoid arthritis. J Clin Med 2024; 13: 5057.
7. Zhou Z, Sun B, Yu D, Zhu C. Gut microbiota: an important player in type 2 diabetes mellitus. Front Cell Infect Microbiol 2022; 12: 834485.
8. Dmytriv TR, Storey KB, Lushchak VI. Intestinal barrier permeability: the influence of gut microbiota, nutrition, and exercise. Front Physiol 2024; 15: 1380713.
9. Peña-Durán E, García-Galindo JJ, López-Murillo LD, Huerta-Huerta A, Balleza-Alejandri LR, Beltrán-Ramírez A, et al. Microbiota and inflammatory markers: a review of their interplay, clinical implications, and metabolic disorders. Int J Mol Sci 2025; 26: 1773.
10. Ejtahed HS, Hoseini-Tavassol Z, Khatami S, Zangeneh M, Behrouzi A, Ahmadi Badi S, et al. Main gut bacterial composition differs between patients with type 1 and type 2 diabetes and non-diabetic adults. J Diabetes Metab Disord 2020; 19: 265-271.
11. Slouha E, Rezazadah A, Farahbod K, Gerts A, Clunes LA, Kollias TF. Type-2 diabetes mellitus and the gut microbiota: Systematic review. Cureus 2023; 15(11): e49740.
12. Noori P, Sotoodehnejadnematalahi F, Rahimi P, Siadat SD. Akkermansia muciniphila and its extracellular vesicles affect Endocannabinoid system in in vitro model. Digestion 2025; 106: 338-348.
13. Tanase DM, Gosav EM, Neculae E, Costea CF, Ciocoiu M, Hurjui LL, et al. Role of gut microbiota on onset and progression of microvascular complications of type 2 diabetes (T2DM). Nutrients 2020; 12: 3719.
14. Fliegerová KO, Mahayri TM, Sechovcová H, Mekadim C, Mrázek J, Jarošíková R, et al. Diabetes and gut microbiome. Front Microbiol 2025; 15: 1451054.
15. Zhao S, Xiang J, Abedin M, Wang J, Zhang Z, Zhang Z, et al. Characterization and anti-inflammatory effects of Akkermansia muciniphila-derived extracellular vesicles. Microorganisms 2025; 13: 464.
16. Koval S, Snihurska I. Role of gut microbiota in the pathogenesis of type 2 diabetes mellitus (literature review). Med Perspekt 2021; 26: 22-30.
17. Wu J, Yang K, Fan H, Wei M, Xiong Q. Targeting the gut microbiota and its metabolites for type 2 diabetes mellitus. Front Endocrinol (Lausanne) 2023; 14: 1114424.
18. Carding S, Verbeke K, Vipond DT, Corfe BM, Owen LJ. Dysbiosis of the gut microbiota in disease. Microb Ecol Health Dis 2015; 26: 26191.
19. Liu Y, Lou X. Type 2 diabetes mellitus-related environmental factors and the gut microbiota: emerging evidence and challenges. Clinics (Sao Paulo) 2020; 75: e1277.
20. Fong W, Li Q, Yu J. Gut microbiota modulation: a novel strategy for prevention and treatment of colorectal cancer. Oncogene 2020; 39: 4925-4943.
21. Sedighi M, Razavi S, Navab-Moghadam F, Khamseh ME, Alaei-Shahmiri F, Mehrtash A, et al. Comparison of gut microbiota in adult patients with type 2 diabetes and healthy individuals. Microb Pathog 2017; 111: 362-369.
22. Candela M, Biagi E, Soverini M, Consolandi C, Quercia S, Severgnini M, et al. Modulation of gut microbiota dysbioses in type 2 diabetic patients by macrobiotic Ma-Pi 2 diet. Br J Nutr 2016; 116: 80-93.
23. Lê KA, Li Y, Xu X, Yang W, Liu T, Zhao X, et al. Alterations in fecal Lactobacillus and Bifidobacterium species in type 2 diabetic patients in Southern China population. Front Physiol 2013; 3: 496.
24. Wu T, Zhang Y, Li W, Zhao Y, Long H, Muhindo EM, et al. Lactobacillus rhamnosus LRa05 ameliorate hyperglycemia through a regulating glucagon-mediated signaling pathway and gut microbiota in type 2 diabetic mice. J Agric Food Chem 2021; 69: 8797-8806.
25. Gulnaz A, Nadeem J, Han J-H, Lew L-C, Son J-D, Park Y-H, et al. Lactobacillus SPS in reducing the risk of diabetes in high-fat diet-induced diabetic mice by modulating the gut microbiome and inhibiting key digestive enzymes associated with diabetes. Biology (Basel) 2021; 10: 348.
26. Remely M, Hippe B, Zanner J, Aumueller E, Brath H, Haslberger AG. Gut microbiota of obese, type 2 diabetic individuals is enriched in Faecalibacterium prausnitzii, Akkermansia muciniphila and Peptostreptococcus anaerobius after weight loss. Endocr Metab Immune Disord Drug Targets 2016; 16: 99-106.
27. Qin J, Li Y, Cai Z, Li S, Zhu J, Zhang F, et al. A metagenome-wide association study of gut microbiota in type 2 diabetes. Nature 2012; 490: 55-60.
28. El-Sabbagh A, El Sayed Zaki M, Motawea MM, Alkasaby NM. Molecular study of lactobacilli species in patients with Type 2 Diabetes Mellitus. Open Microbiol J 2022; 16: 10.2174/18742858-v16-e2205090.
29. Larsen N, Vogensen FK, van den Berg FW, Nielsen DS, Andreasen AS, Pedersen BK, et al. Gut microbiota in human adults with type 2 diabetes differs from non-diabetic adults. PloS One 2010; 5(2): e9085.
30. Tsai HJ, Tsai YC, Hung WW, Hung WC, Chang CC, Dai CY. Gut microbiota and non-alcoholic fatty liver disease severity in type 2 diabetes patients. J Pers Med 2021; 11: 238.
31. Ebrahimzadeh Leylabadlo H, Ghotaslou R, Samadi Kafil H, Feizabadi MM, Moaddab SY, Farajnia S, et al. Non-alcoholic fatty liver diseases: from role of gut microbiota to microbial-based therapies. Eur J Clin Microbiol Infect Dis 2020; 39: 613-627.
32. Murri M, Leiva I, Gomez-Zumaquero JM, Tinahones FJ, Cardona F, Soriguer F, et al. Gut microbiota in children with type 1 diabetes differs from that in healthy children: a case-control study. BMC Med 2013; 11: 46.
33. Zhang J, Ni Y, Qian L, Fang Q, Zheng T, Zhang M, et al. Decreased abundance of Akkermansia muciniphila leads to the impairment of insulin secretion and glucose homeostasis in lean type 2 diabetes. Adv Sci (Weinh) 2021; 8(16): e2100536.
34. Zhang L, Qin Q, Liu M, Zhang X, He F, Wang G. Akkermansia muciniphila can reduce the damage of gluco/lipotoxicity, oxidative stress and inflammation, and normalize intestine microbiota in streptozotocin-induced diabetic rats. Pathog Dis 2018; 76: 10.1093/femspd/fty028.
35. Sanjiwani MID, Aryadi IPH, Semadi IMS. Review of literature on Akkermansia muciniphila and its possible role in the etiopathogenesis and therapy of type 2 diabetes mellitus. J ASEAN Fed Endocr Soc 2022; 37: 69-74.
36. Rodrigues VF, Elias-Oliveira J, Pereira ÍS, Pereira JA, Barbosa SC, Machado MSG, et al. Akkermansia muciniphila and gut immune system: a good friendship that attenuates inflammatory bowel disease, obesity, and diabetes. Front Immunol 2022; 13: 934695.
| Files | ||
| Issue | Vol 17 No 6 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i6.20354 | |
| Keywords | ||
| Diabetes mellitus Gut microbiota Real-time polymerase chain reaction Case-control | ||
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How to Cite
1.
Saiedi E, Shapouri R, Haghi F, Zeighami H. Comparative analysis of gut microbiota composition in the fecal samples from type 2 diabetes mellitus patients and healthy individuals: a case control study. Iran J Microbiol. 2025;17(6):875-884.
