Predictive association of gut microbiome and NLR in anemic low middle-income population of Odisha- a cross-sectional study

BackgroundIron is abundant on earth but not readily available for colonizing bacteria due to its low solubility in the human body. Hosts and microbiota compete fiercely for iron. <15% Supplemented Iron is absorbed in the small bowel, and the remaining iron is a source of dysbiosis. The gut microbiome signatures to the level of predicting anemia among low-middle-income populations are unknown. The present study was conducted to identify gut microbiome signatures that have predictive potential in association with Neutrophil to lymphocytes ratio (NLR) and Mean corpuscular volume (MCV) in anemia.MethodsOne hundred and four participants between 10 and 70 years were recruited from Odisha’s Low Middle-Income (LMI) rural population. Hematological parameters such as Hemoglobin (HGB), NLR, and MCV were measured, and NLR was categorized using percentiles. The microbiome signatures were analyzed from 61 anemic and 43 non-anemic participants using 16 s rRNA sequencing, followed by the Bioinformatics analysis performed to identify the diversity, correlations, and indicator species. The Multi-Layered Perceptron Neural Network (MLPNN) model were applied to predict anemia.ResultsSignificant microbiome diversity among anemic participants was observed between the lower, middle, and upper Quartile NLR groups. For anemic participants with NLR in the lower quartile, alpha indices indicated bacterial overgrowth, and consistently, we identified R. faecis and B. uniformis were predominating. Using ROC analysis, R. faecis had better distinction (AUC = 0.803) to predict anemia with lower NLR. In contrast, E. biforme and H. parainfluenzae were indicators of the NLR in the middle and upper quartile, respectively. While in Non-anemic participants with low MCV, the bacterial alteration was inversely related to gender. Furthermore, our Multi-Layered Perceptron Neural Network (MLPNN) models also provided 89% accuracy in predicting Anemic or Non-Anemic from the top 20 OTUs, HGB level, NLR, MCV, and indicator species.ConclusionThese findings strongly associate anemic hematological parameters and microbiome. Such predictive association between the gut microbiome and NLR could be further evaluated and utilized to design precision nutrition models and to predict Iron supplementation and dietary intervention responses in both community and clinical settings

Common NOD2 mutations are absent in patients with Crohn's disease in India

Background: Crohn's disease is being increasingly diagnosed in the Indian subcontinent. Three apparently common mutations in the NOD2 gene are found in up to 30% of sporadic patients with Crohn's disease in western countries. We examined whether such mutations are also found in Indian patients with Crohn's disease. Methods: Venous blood was collected from 82 patients (age range: 7-65 years, 53 men) with Crohn's disease and 149 control subjects; DNA was extracted and subjected to polymerase chain reaction using specific primers. The amplified fragments of size 185, 163 and 151 bp for R702W, G908R and 1007fs, respectively, were digested with MspI, HhaI and ApaI, and the restriction pattern noted after electrophoresis. Results: Twenty-eight patients had ileocolonic disease, 26 ileal disease, 20 colonic disease and 8 had disease limited to proximal small bowel or stomach. None of the 82 patients showed any of the three NOD2 mutations. The control subjects (93 men) had a variety of chronic gastrointestinal disorders (ulcerative colitis 52, irritable bowel syndrome 30, intestinal tuberculosis 20, colon cancer 7, miscellaneous 37). None of the control subjects showed a mutation in any of the three NOD2 mutation analyses. Conclusion: The three NOD2 gene mutations described above are uncommon in Indian patients with Crohn's disease. This study complements information provided by recent studies on NOD2 mutations in Indians

Bacterial succession in the colon during childhood and adolescence: molecular studies in a Southern Indian village

Background: The colonic bacterial flora, largely anaerobic, is believed to establish and stabilize in the first 2 y of life. Objective: This study was undertaken to determine whether the bacterial flora of the colon undergoes further changes (succession) during childhood and adolescence. Design: This cross-sectional study examined fecal samples from 130 healthy children and adolescents in the age group 2-17 y and from 30 healthy adults (median age: 42 y) residing in a single village in southern India. DNA was extracted and subjected to 16S rDNA-targeted real-time polymerase chain reaction to determine the relative predominance of Bifidobacterium genus, Bacteroides-Prevotella-Porphyromonas group, Lactobacillus acidophilus group, Eubacterium rectale, and Faecalibacterium prausnitzii. Results: Bifidobacterium species and Bacteroides-Prevotella group were dominant fecal bacteria overall. E. rectale and Lactobacillus species were considerably less abundant. Clear age-related differences emerged, with a steep decline in Bifidobacterium species in adults (P < 0.0001), a steep decline of Lactobacillus species >5 y of age (P < 0.0001), an increase in Bacteroides during late adolescence and in adults (P = 0.0040), an increase in E. rectale during childhood and adolescence followed by a steep decline in adults (P < 0.0001), and a late childhood peak of F. prausnitzii with decline in adolescents and adults (P < 0.0001). Conclusions: Changes in the bacterial flora occur during childhood and adolescence characterized by reduction in Lactobacillus and Bifidobacterium species and an increase in Bacteroides, E rectale, and F. prausnitzii peaked during late childhood in this population

Faecal bifidobacteria in Indian neonates & the effect of asymptomatic rotavirus infection during the first month of life

Background & objectives: Bifidobacteria colonize the gut after the first week of life and remain an important component of the gut microbiota in infancy. This study was carried out to characterize the diversity and number of bifidobacteria colonizing the gut in Indian neonates and to investigate whether asymptomatic infection with rotavirus in the first month of life affected gut colonization by bidifobacteria. Methods: DNA was isolated from faeces of 14 term-born neonates who were under surveillance for rotavirus infection. Bacterial and bifidobacterial diversity was evaluated by temporal temperature gradient electrophoresis (TTGE) of 16S rDNA amplified using total bacteria and bifidobacteria-specific primers. Real time PCR, targeting 16S rDNA, was used to quantitate faecal bifidobacteria and enterobacteria. Results: TTGE of conserved bacterial 16S rDNA showed 3 dominant bands of which Escherichia coli (family Enterobacteriaceae) and Bifidobacterium (family Bifidobacteriaceae) were constant. TTGE of Bifidobacterium genus-specific DNA showed a single band in all neonates identified by sequencing as Bifidobacterium longum subsp. infantis. Faecal bifidobacterial counts (log10 cfu/g faeces) ranged from 6.1 to 9.3 and enterobacterial counts from 6.3 to 9.5. Neonates without and with rotavirus infection in the first week of life did not show significant differences in the median count of bifidobacteria (log10 count 7.48 vs. 7.41) or enterobacteria (log10 count 8.79 vs. 7.92). Interpretation amp; conclusions: B. longum subsp. infantis was the sole bifidobacterial species colonizing the gut of Indian neonates. Asymptomatic rotavirus infection in the first month of life was not associated with alteration in faecal bifidobacteria or enterobacteria

Molecular studies of fecal anaerobic commensal bacteria in acute diarrhea in children

Background and Objective: The commensal bacterial flora of the colon may undergo changes during diarrhea, owing to colonization of the intestine by pathogens and to rapid intestinal transit. This study used molecular methods to determine changes in the composition of selected commensal anaerobic bacteria during and after acute diarrhea in children. Materials and Methods: Fecal samples were obtained from 46 children with acute diarrhea in a rural community during an episode of acute diarrhea, immediately after recovery from diarrhea, and 3 months after recovery. DNA was extracted and quantitative polymerase chain reaction using SYBR green and genus- and species-specific primers targeting 16S rDNA were undertaken to quantitate the following groups of bacteria: Bifidobacterium spp., Bifidobacterium longum group, Bacteroides-Prevotella group, Bacteroides fragilis, Lactobacillus acidophilus group, Faecalibacterium prauznitzii, and Eubacterium rectale, relative to amplification of universal bacterial domain 16S rDNA. Results: Bacteria belonging to the Bacteroides-Prevotella-Porphyromonas group, E rectale, L acidophilus, and F prauznitzii groups were low during acute diarrhea compared with their levels after recovery from diarrhea. The pattern was similar in rotavirus diarrhea and nonrotavirus diarrhea. Administration of amylase-resistant maize starch as adjuvant therapy was associated with lower levels of F prauznitzii at the time of recovery but did not lead to other changes in the floral pattern. Conclusions: Specific classes of fecal bacteria are lower during episodes of acute diarrhea in children than during periods of normal gastrointestinal health, suggesting specific alterations in the flora during diarrhea

Drug resistant epilepsy and ketogenic diet: A narrative review of mechanisms of action

Drug-resistant epilepsy (DRE) poses a significant global challenge, impacting the well-being of patients. Anti-epileptic drugs often fail to effectively control seizures in individuals with DRE. This condition not only leads to persistent seizures but also induces neurochemical imbalances, elevating the risk of sudden unexpected death in epilepsy and comorbidities. Moreover, patients experience mood and personality alterations, educational and vocational setbacks, social isolation, and cognitive impairments. Ketogenic diet has emerged as a valuable therapeutic approach for DRE, having been utilized since 1920. Various types of ketogenic diets have demonstrated efficacy in controlling seizures. By having a multimodal mechanism of action, the ketogenic diet reduces neuronal excitability and the frequency of seizure episodes. In our narrative review, we have initially provided a concise overview of the factors contributing to drug resistance in epilepsy. Subsequently, we have discussed the different available ketogenic diets. We have reviewed the underlying mechanisms through which the ketogenic diet operates. These mechanisms encompass decreased neuronal excitability, enhanced mitochondrial function, alterations in sleep patterns, and modulation of the gut microbiome. Understanding the complex mechanisms by which this diet acts is essential as it is a rigorous diet and requires good compliance. Hence knowledge of the mechanisms may help to advance research on achieving similar therapeutic effects through other less stringent approaches

Fecal polymerase chain reaction for Mycobacterium tuberculosis IS6110 to distinguish Crohn's disease from intestinal tuberculosis

Background: We have previously shown that amplification of Mycobacterium tuberculosis specific DNA (TB PCR) from feces reliably diagnosed intestinal tuberculosis. This study was undertaken to determine how well this test would distinguish intestinal tuberculosis from Crohn's disease in a country endemic for tuberculosis. Methods: Consecutive patients with diagnoses of Crohn's disease and intestinal tuberculosis were enrolled, and the diagnoses confirmed by follow up. DNA was extracted from fecal samples and subjected to polymerase chain reaction TB PCR for IS6110 sequence which is specific for M. tuberculosis. Results: Twenty one of 24 patients with intestinal tuberculosis and 5 of 44 patients with Crohn's disease tested positive by TB PCR. The sensitivity, specificity, positive predictive and negative predictive values for TB PCR in distinguishing tuberculosis from Crohn's disease were 0.79 (95% confidence interval 0.57-0.92), 0.88 (0.75-0.96), 0.79 (0.57-0.92) and 0.88 (0.75-0.96), respectively. A combination of fecal TB PCR with mycobacterial culture of mucosal biopsy specimens identified 23 of 24 (96.2%) of patients with intestinal TB, with sensitivity, specificity, positive predictive and negative predictive values (95% CI) of 0.95 (0.78-0.99), 0.88 (0.75-0.96), 0.82 (0.63-0.93) and 0.97 (0.86-0.99), respectively. Conclusion Fecal TB PCR is a good screening test to distinguish intestinal tuberculosis from Crohn's disease

Supplementation of higher doses of Iron in programmes to control anaemia is a double edged sword

Anemia is a major public health problem globally. Due to loss of 42.2 million disability-adjusted life years (DALYs) in 2011, Iron-deficiency anemia (IDA) was ranked among the top three major causes of disability in the world (1,2). IDA is responsible for at least 50% of anemia and is highly prevalent in India especially among women, children below 5 years, and adolescent girls (3) perhaps due to increased needs, insufficient dietary supplies, poor intervention coverage etc (4). Anemic girls will grow up to anemic mothers and in turn give birth to anemic children. Childhood anemia can have long term consequences. IDA has been associated with developmental deficits, impaired memory and neurodevelopment, diminished physical function, depression, fatigue, loss of vitality, preterm delivery, and lower infant birth weight (5–10). These, effects attributable to anemia remain invisible but are substantial (11), as it has serious health and economic costs and may hinder nation’s development. Therefore, effective and safe interventions are urgently needed across the lifecycle. The issue has attracted global attention and for the first time targets have been suggested like reducing the proportion of anemia among women of reproductive age (WRA) by 50% by 2025 in comparison to the baseline year 2011 (12). This is likely to add impetus to global efforts in reducing anemia and monitor the progress towards it