Alpha Diversity Analysis of Microbiota Dysbiosis in Normal and Colorectal Cancer of Mice Feces
DOI:
https://doi.org/10.6000/1929-6029.2025.14.46Keywords:
Colorectal cancer, gut microbiota, Next Generation Sequencing, dysbiosis, alpha diversityAbstract
Background: Colorectal cancer development is influenced by both environmental and genetic factors, with the gut microbiota playing a significant role. This research investigates how alterations in gut microbiota are associated with the incidence, progression, prognosis, and early detection of CRC.
Methods: An experimental laboratory study was carried out using Sprague Dawley rats that were induced with azoxymethane (AOM) and Dextran Sodium Sulfate (DSS). The thirty rats were divided into three groups: normal, cancer-induced, and treatment. The fecal microbiota profiles were examined through Next Generation Sequencing (NGS), and the data were analyzed for alpha diversity, highlighting the dynamics of the microbial community.
Results: The cancer-induced group (K2 Plus) exhibited the highest microbial diversity across Shannon, Simpson, Chao1, and PD Whole Tree indices, while the treatment group (P2 Plus) demonstrated the lowest.
Conclusion: These findings suggest that the increase in diversity observed in cancer-induced mice reflects disruption of community stability and blooming of pathobionts. Conversely, treatment with Lactococcus lactis D4 reduced diversity, potentially by selectively suppressing pro-inflammatory or pathogenic taxa, indicating a beneficial probiotic effect in mitigating dysbiosis associated with colorectal cancer.
References
Hossain MS, Karuniawati H, Jairoun AA, et al. Colorectal Cancer: A Review of Carcinogenesis, Global Epidemiology, Current Challenges, Risk Factors, Preventive and Treatment Strategies. Cancers (Basel) 2022; 14. DOI: https://doi.org/10.3390/cancers14071732
Gupta A, Madani R, Mukhtar H. Streptococcus bovis endocarditis, a silent sign for colonic tumour. Colorectal Dis 2010; 12: 164-71. DOI: https://doi.org/10.1111/j.1463-1318.2009.01814.x
Kostic AD, Gevers D, Pedamallu CS, et al. Genomic analysis identifies association of Fusobacterium with colorectal carcinoma. Genome Res 2012; 22: 292-8. DOI: https://doi.org/10.1101/gr.126573.111
Viljoen KS, Dakshinamurthy A, Goldberg P, et al. Quantitative profiling of colorectal cancer-associated bacteria reveals associations between fusobacterium spp, enterotoxigenic Bacteroides fragilis (ETBF) and clinicopathological features of colorectal cancer. PLoS One 2015. 10: p. e0119462. DOI: https://doi.org/10.1371/journal.pone.0119462
Buc E, Dubois D, Sauvanet P, et al. High prevalence of mucosa-associated E. coli producing cyclomodulin and genotoxin in colon cancer. PLoS One 2013; 8: e56964. DOI: https://doi.org/10.1371/journal.pone.0056964
Wang T, Cai G, Qiu Y, et al. Structural segregation of gut microbiota between colorectal cancer patients and healthy volunteers. ISME J 2012; 6: 320-9. DOI: https://doi.org/10.1038/ismej.2011.109
Porter RJ, Arends MJ, Churchhouse AMD, et al. Inflammatory Bowel Disease-Associated Colorectal Cancer: Translational Risks from Mechanisms to Medicines. J Crohns Colitis 2021; 15: 2131-2141. DOI: https://doi.org/10.1093/ecco-jcc/jjab102
Lucafò M, Curci D, Franzin M, et al. Inflammatory Bowel Disease and Risk of Colorectal Cancer: An Overview From Pathophysiology to Pharmacological Prevention. Front Pharmacol 2021; 12: 772101. DOI: https://doi.org/10.3389/fphar.2021.772101
Nardone OM, Zammarchi I, Santacroce G, et al. Inflammation-Driven Colorectal Cancer Associated with Colitis: From Pathogenesis to Changing Therapy. Cancers (Basel) 2023; 15. DOI: https://doi.org/10.3390/cancers15082389
Tanaka T, Kohno H, Suzuki R, et al. A novel inflammation-related mouse colon carcinogenesis model induced by azoxymethane and dextran sodium sulfate. Cancer Sci 2003; 94: 965-73. DOI: https://doi.org/10.1111/j.1349-7006.2003.tb01386.x
Tian R, Zuo X, Jaoude JC, et al. ALOX15 as a Suppressor of Inflammation and Cancer: Lost in the Link. Prostaglandins & Other Lipid Mediators 2017. DOI: https://doi.org/10.1016/j.prostaglandins.2017.01.002
Ohnishi S, Hiramoto K, Ma N, et al. Chemoprevention by Aspirin Against Inflammation-Related Colorectal Cancer in Mice. Journal of Clinical Biochemistry and Nutrition 2021. DOI: https://doi.org/10.3164/jcbn.20-189
Yang X, Wang Q, Zhang X, et al. Purple Yam Polyphenol Extracts Exert Anticolitis and Anticolitis-Associated Colorectal Cancer Effects Through Inactivation of NF-κB/p65 and STAT3 Signaling Pathways. Journal of Agricultural and Food Chemistry 2023. DOI: https://doi.org/10.1021/acs.jafc.3c00346
Stolfi C, Rizzo A, Franzè E, et al. Involvement of interleukin-21 in the regulation of colitis-associated colon cancer. Journal of Experimental Medicine 2011; 208: 2279-2290. DOI: https://doi.org/10.1084/jem.20111106
Zhang B, Xu Y, Liu S, et al. Dietary Supplementation of Foxtail Millet Ameliorates Colitis-Associated Colorectal Cancer in Mice via Activation of Gut Receptors and Suppression of the STAT3 Pathway. Nutrients 2020; 12. DOI: https://doi.org/10.3390/nu12082367
Ock CY, Kim E-H, Hong H, et al. Prevention of Colitis-Associated Colorectal Cancer with 8-Hydroxydeoxyguano-sine. Cancer Prevention Research 2011; 4: 1507-1521. DOI: https://doi.org/10.1158/1940-6207.CAPR-11-0161
Meana C, García-Rostán G, Peña L, et al. The phosphatidic acid phosphatase lipin-1 facilitates inflammation-driven colon carcinogenesis. JCI Insight 2018; 3. DOI: https://doi.org/10.1172/jci.insight.97506
Pandurangan AK, Saadatdoust Z, Mohd NE, et al. Dietary cocoa protects against colitis-associated cancer by activating the Nrf2/Keap1 pathway. BioFactors 2015; 41: 1-14. DOI: https://doi.org/10.1002/biof.1195
Khalyfa AA, Punatar S, Aslam R, et al. Exploring the Inflammatory Pathogenesis of Colorectal Cancer. Diseases 2021; 9. DOI: https://doi.org/10.3390/diseases9040079
Rosenberg E, DeLong EF, Thompson F, et al. The prokaryotes: Human Microbiology 2013; 1-554. DOI: https://doi.org/10.1007/978-3-642-30144-5
Cheng Y, Ling Z, Li L. The Intestinal Microbiota and Colorectal Cancer. Front Immunol 2020; 11: 615056. DOI: https://doi.org/10.3389/fimmu.2020.615056
Rausch P, Rühlemann M, Hermes BM, et al. Comparative analysis of amplicon and metagenomic sequencing methods reveals key features in the evolution of animal metaorganisms. Microbiome 2019; 7: 133. DOI: https://doi.org/10.1186/s40168-019-0743-1
Heidrich V, Inoue LT, Asprino PF, et al. Choice of 16S Ribosomal RNA Primers Impacts Male Urinary Microbiota Profiling. Front Cell Infect Microbiol 2022; 12: 862338. DOI: https://doi.org/10.3389/fcimb.2022.862338
Hoffman C, Siddiqui NY, Fields I, et al. Species-Level Resolution of Female Bladder Microbiota from 16S rRNA Amplicon Sequencing. mSystems 2021; 6: e0051821. DOI: https://doi.org/10.1128/msystems.00518-21
Höyhtyä M, Korpela K, Saqib S, et al. Quantitative Fecal Microbiota Profiles Relate to Therapy Response During Induction With Tumor Necrosis Factor α Antagonist Infliximab in Pediatric Inflammatory Bowel Disease. Inflamm Bowel Dis 2023; 29: 116-124. DOI: https://doi.org/10.1093/ibd/izac182
Zhu Q, Huang S, Gonzalez A, et al. Phylogeny-Aware Analysis of Metagenome Community Ecology Based on Matched Reference Genomes while Bypassing Taxonomy. mSystems 2022; 7: e00167-22. DOI: https://doi.org/10.1128/msystems.00167-22
Peterson D, Bonham KS, Rowland S, et al. Comparative Analysis of 16S rRNA Gene and Metagenome Sequencing in Pediatric Gut Microbiomes. Front Microbiol 2021; 12: 670336. DOI: https://doi.org/10.3389/fmicb.2021.670336
Jovel J, Patterson J, Wang W, et al. Characterization of the Gut Microbiome Using 16S or Shotgun Metagenomics. Front Microbiol 2016; 7: 459. DOI: https://doi.org/10.3389/fmicb.2016.00459
Poretsky R, Rodriguez RL, Luo C, et al. Strengths and limitations of 16S rRNA gene amplicon sequencing in revealing temporal microbial community dynamics. PLoS One 2014; 9: e93827. DOI: https://doi.org/10.1371/journal.pone.0093827
Rezasoltani S, Ahmadi Bashirzadeh D, Nazemalhosseini Mojarad E, et al. Signature of Gut Microbiome by Conventional and Advanced Analysis Techniques: Advantages and Disadvantages. Middle East J Dig Dis 2020; 12: 5-11. DOI: https://doi.org/10.15171/mejdd.2020.157
Zheng X, Zhu Q, Qin M, et al. The Role of Feeding Characteristics in Shaping Gut Microbiota Composition and Function of Ensifera (Orthoptera). Insects 2022; 13. DOI: https://doi.org/10.3390/insects13080719
Sharpton TJ. An introduction to the analysis of shotgun metagenomic data. Front Plant Sci 2014; 5: 209. DOI: https://doi.org/10.3389/fpls.2014.00209
Willis AD. Rarefaction, Alpha Diversity, and Statistics. Front Microbiol 2019; 10: 2407. DOI: https://doi.org/10.3389/fmicb.2019.02407
Moreno C, Barragan F, Pineda E, et al. Reanálisis de la diversidad alfa: alternativas para interpretar y comparar información sobre comunidades ecológicas. Revista Mexicana de Biodiversidad 2011; 82: 1249-1261. DOI: https://doi.org/10.22201/ib.20078706e.2011.4.745
McCoy CO, Matsen FAT Abundance-weighted phylogenetic diversity measures distinguish microbial community states and are robust to sampling depth. Peer J 2013; 1: e157. DOI: https://doi.org/10.7717/peerj.157
Walters KE, Martiny JBH. Alpha-, beta-, and gamma-diversity of bacteria varies across habitats. PLoS One 2020; 15: e0233872. DOI: https://doi.org/10.1371/journal.pone.0233872
Contoli L. Diversity Indices as ‘Magic’ Tools in Landscape Planning: A Cautionary Note on their Uncritical Use. Landscape Research 2011; 36: 111-117. DOI: https://doi.org/10.1080/01426397.2010.535896
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