Stool Samples Research Articles

Associations between brain morphology, inflammatory markers and symptoms of fatigue, depression or anxiety in active and remitted Crohn's disease.

Fatigue and psychosocial impairments are highly prevalent in IBD, especially during active disease. Disturbed brain-gut-interactions may contribute to these symptoms. This study examined associations between brain structure, faecal calprotectin and symptoms of fatigue, depression and anxiety in persons with Crohn's Disease (CD) in different disease states. In this prospective observational study, n=109 participants (n=67 persons with CD, n=42 healthy controls) underwent cranial magnetic resonance imaging, provided stool samples for analysis of faecal calprotectin and completed questionnaires to assess symptoms of fatigue, depression and anxiety. We analysed differences in grey matter volume (GMV) between patients and controls and associations between regional GMV alterations, neuropsychiatric symptoms and faecal calprotectin. Symptoms of fatigue, depression and anxiety were increased in patients with CD compared to controls, with highest scores in active CD. Patients exhibited regionally reduced GMV in cortical and subcortical sensorimotor regions, occipitotemporal and medial frontal areas. Regional GMV differences showed a significant negative association with fatigue, but not with depression or anxiety. Subgroup analyses revealed symptom-GMV-associations for fatigue in remitted, but not in active CD, while fatigue was positively associated with faecal calprotectin in active, but not remitted disease. Our findings support disturbed brain-gut-interactions in CD which may be particularly relevant for fatigue during remitted disease. Reduced GMV in the precentral gyrus and other sensorimotor areas could reflect key contributions to fatigue pathophysiology in CD. A sensorimotor model of fatigue in CD could also pave the way for novel treatment approaches.

Prospective association of the infant gut microbiome with social behaviors in the ECHO consortium.

Identifying modifiable risk factors of autism spectrum disorders (ASDs) may inform interventions to reduce financial burden. The infant/toddler gut microbiome is one such feature that has been associated with social behaviors, but results vary between cohorts. We aimed to identify consistent overall and sex-specific associations between the early-life gut microbiome and autism-related behaviors. Utilizing the Environmental influences on Children Health Outcomes (ECHO) consortium of United States (U.S.) pediatric cohorts, we gathered data on 304 participants with fecal metagenomic sequencing between 6-weeks to 2-years postpartum (481 samples). ASD-related social development was assessed with the Social Responsiveness Scale (SRS-2). Linear regression, PERMANOVA, and Microbiome Multivariable Association with Linear Models (MaAsLin2) were adjusted for sociodemographic factors. Stratified models estimated sex-specific effects. Genes encoding pathways for synthesis of short-chain fatty acids were associated with higher SRS-2 scores, indicative of ASDs. Fecal concentrations of butyrate were also positively associated with ASD-related SRS-2 scores, some of which may be explained by formula use. The distribution of age at outcome assessment differed in the cohorts included, potentially limiting comparability between cohorts. Stool sample collection methods also differed between cohorts. Our study population reflects the general U.S. population, and thus includes few participants who met the criteria for being at high risk of developing ASD. Our study is among the first multicenter studies in the U.S. to describe prospective microbiome development from infancy in relation to neurodevelopment associated with ASDs. Our work contributes to clarifying which microbial features associate with subsequent diagnosis of neuropsychiatric outcomes. This will allow for future interventional research targeting the microbiome to change neurodevelopmental trajectories.

Rapid Management of Serratia marcescens Outbreak in Neonatology Unit in Singapore: Risk factors and Infection Control Measures.

To describe two consecutive Serratia marcescens outbreaks in neonatology unit of Singapore General hospital and determine risk factors associated with its acquisition in the second outbreak. Epidemiological investigations, environmental sampling and risk-factors analysis were used to guide infection control measures. Active surveillance sampling of nasopharyngeal aspirate and/or stool from neonates was conducted during both outbreaks. Whole-genome-sequencing was performed to determine clonal links. Retrospective case-control study was conducted in the second outbreak to identify risk factors for S. marcescens acquisition. In the one-year period in 2022, two S. marcescens outbreaks were managed involving five neonates in March 2022 outbreak and eight neonates in November 2022 outbreak. Both outbreaks were caused by genetically unrelated S. marcescens clones, with a link to positive isolates from sinks in intensive care units and milk preparation room during outbreak 1. Neonatal jaundice(OR, 16.46; p-value= 0.023) and non-formula milk feeding(OR, 13.88; p-value= 0.02) were identified as independent risk factors during second outbreak. Multiple interventions adopted were cohorting of positive cases, carriage-screening, enhanced environmental cleaning, and emphasis on alcohol-based handrubs for hand-hygiene. The two outbreaks in our institution were likely due to infection prevention practices lapses and favourable environmental conditions. Nosocomial S. marcescens outbreaks in neonatology units are difficult to control and require multidisciplinary approach with strict infection prevention measures to mitigate infection risks.

Magnitude of intestinal parasitic infections and its determinants among HIV/AIDS patients attending at antiretroviral treatment centers in East and West Gojam Zones, Northwest, Ethiopia: institution based cross-sectional study.

Intestinal parasitic infections (IP) are a major source of morbidity in people living with Human immunodeficiency virus (HIV), particularly in resource-limited settings, mostly as a result of high viral load. Hence, this study aimed to investigate the magnitude of intestinal parasitic infections and its determinants among patients with HIV/AIDS attending public health facilities in East and West Gojam Zones in Ethiopia. Institution-based cross-sectional study was conducted on 327people living with HIV visiting public health facilities from December 2022 to May 2023. A simple random sampling technique was used to recruit participants. Face-to-face interviews were used to collect socio-demographics and determinants. The fresh stool was collected from each patient, transported, and tested in accordance with laboratory standard operating procedures of wet mount, formol-ether concentration technique, and modified acid-fast staining. Data were entered and analyzed in the statistical package for Social Science (SPSS) version 20. A 95% CI with p-value < 0.05 was considered statistically significant. The overall prevalence of IP in patients with HIV/AIDS was 19.3% (63/327). Hookworm was the most identified parasite 33.3% (21/63) followed by E.histolytica 17% (11/63) and G.lamblia 14.3% (9/63). Parasitic infections were significantly higher among viral load > 1000cps/ml (p = 0.035), WHO stage 4 (p = 0.002), CD4 < 200cell/mm3 (p = 0.001), and bare foot walking (p = 0.001). IP infections are moderately high among patients with HIV/AIDS in the study area. The proportion of parasites was greatly affected by high viral load, WHO stage 4, CD4 < 200cell/mm3, and being barefoot; this gives valuable insight to health professionals, health planners and community health workers. As a result, viral load monitoring, and WHO stage controlling were periodically assessed in patients with HIV/AIDS. Health education, awareness creation, routine stool examination, and environmental hygiene were regularly advocated to increase the life of patients with HIV/AIDS.

Structural and functional alteration of the gut microbiota in elderly patients with hyperlipidemia

ObjectiveTo investigate the structure, composition, and functions of the gut microbiota in elderly patients with hyperlipidemia.MethodsSixteen older patients diagnosed with hyperlipidemia (M group) and 10 healthy, age-matched normal volunteers (N group) were included. These groups were further subdivided by sex into the male normal (NM, n = 5), female normal (NF, n = 5), male hyperlipidemia (MM, n = 8), and female hyperlipidemia (MF, n = 8) subgroups. Stool samples were collected for high-throughput sequencing of 16S rRNA genes. Blood samples were collected for clinical biochemical index testing.ResultsAlpha- and beta-diversity analyses revealed that the structure and composition of the gut microbiota were significantly different between the M and N groups. The relative abundances of Bacteroides, Parabacteroides, Blautia, Peptococcus, and Bifidobacterium were significantly decreased, while those of Lactobacillus, Helicobacter, and Desulfovibrio were significantly higher in the M group. There were also significant sex-related differences in microbial structure between the NM and NF groups, and between the MM and MF groups. Through functional prediction with PICRUSt 2, we observed distinct between-group variations in metabolic pathways associated with the gut microbiota and their impact on the functionality of the nervous system. Pearson’s correlation coefficient was used as a distance metric to build co-abundance networks. A hypergeometric test was used to detect taxonomies with significant enrichment in specific clusters. We speculated that modules with Muribaculaceae and Lachnospiraceae as the core microbes play an important ecological role in the intestinal microbiota of the M group. The relative intestinal abundances of Agathobacter and Faecalibacterium in the M group were positively correlated with serum triglyceride and low-density lipoprotein levels, while the relative abundance of Bifidobacterium was negatively correlated with the serum lipoprotein a level.

Distinct gut microbiota signatures associated with progression of atherosclerosis in people living with HIV.

The relationship of microbiota composition dynamics and the progression of subclinical atherosclerosis in people with HIV (PWH) remains unknown. 96-week, prospective, longitudinal study in virologically-suppressed PWH. Carotid intima-media thickness (cIMT) measurements and stool samples were obtained at baseline, 48-week and 96-week visits. cIMT progression was defined as an increase >10% and/or detection of new carotid plaque. To profile the gut microbiome, amplification and sequencing of 16S ribosomal-RNA (V3-V4 variable regions) were carried out following the Illumina protocol. Sequencing was performed with MiSeq platform. 191, 190 and 167 patients had available fecal samples for microbiome analysis at the baseline, 48- and 96-week visits, respectively. 87 (43%) participants showed atherosclerosis progression, and 54 (26.7%) presented new carotid plaque. No significant differences were observed in adjusted α-diversity indices between groups defined by cIMT progression. Beta-diversity determined through principal coordinate analysis distances showed that the groups exhibited distinct microbial profiles (PERMANOVA p-value = 0.03). Longitudinal analysis with ANCOM-BC2 adjusted for traditional cardiovascular risk factors, MSM and nadir CD4 count revealed that cIMT progression was consistently associated with Agathobacter and Ruminococcus_2, while non-progression was consistently associated with Prevotella_7. Progression of atherosclerosis in PWH might be associated with distinctive signatures in the gut microbiota.

Prevalence of Intestinal Parasites among Diabetes Mellitus Patients in Tertiary Care Hospital

Intestinal parasitosis constitutes a serious problem of public health, especially in developing countries. Even though the population of immunocompromised individuals is growing, this is a primary cause of illness and mortality. Numerous studies have demonstrated that immunosuppressive medications, such as those used after transplantation, as well as immune suppression as a primary immunodeficiency in HIV infection, increase the possibility of infection and persistent states of carriage. People whose immune systems were weakened, like in the case of chronic internal disorders or abnormal metabolism, were thought to be more susceptible to infection. A group of metabolic non-communicable diseases known as diabetes mellitus (DM) are characterized by hyperglycemia or elevated blood sugar, which can be brought on by either inadequate secretion of insulin, insulin-resistant cells, or both. Parasitic infections in immunocompromised people are becoming more well-known as major opportunistic microorganisms that cause clinical illnesses. The goal of this research is to see how common intestinal parasite infections are in people with diabetes. The research was carried out at the Chandulal Chandrakar Memorial Medical College, Bhilai, Durg district of Chhattisgarh, India. From March to August 2020, diabetic patients who visited hospitals were studied in this cross-sectional study. This study comprised 188 diabetic individuals with type I or type II diabetes who were taking anti-diabetic medication. With a little spoon and a sterile, dry, leak-proof plastic container with a tight-fitting cover that had the patient’s name and identification number on it, the feces samples were taken. The container was then quickly sent to the parasitology lab of the microbiology department for processing. Stool samples were examined under three headings: macroscopic examination, microscopic examination, and formal ether concentration procedure. A total of 188 DM patients was included with data on socio-demographic characteristics, and from each patient stool sample were collected and processed. Out of which, 27 stool samples were positive for intestinal parasites; showed a prevalence of 14.36%. Female 16 (16.49%) revealed a greater frequency than that of men 11 (12.09%). The age range of 51 to 60 years old exhibited the highest incidence 8 (29.63 %). Stool samples were positive for intestinal parasites with one suffering from a dual infection. Various parasites were detected in which Entamoeba histolytica 10 (5.32%) was most common parasite to isolate followed by Ascaris lumbricoides 9 (4.79%) and Giardia lamblia 6 (3.19%) was least isolated parasite. The most prevalent isolated parasite was Entamoeba histolytica, which was followed by Cryptosporidium parvum. Diabetic patients have an increased risk of parasitic infections, particularly when they have the opportunistic infection Entamoeba histolytica. Patients with diabetes who have been diagnosed with Cryptosporidium parvum have a weaker immune system. Therefore, for the sake of societal welfare, people with diabetes mellitus should have routine stool examination screening for parasite infection.

Multiplex PCR for bacterial, viral and protozoal pathogens in persistent diarrhoea or persistent abdominal pain in Côte d’Ivoire, Mali and Nepal

In contrast to acute diarrhoea, the aetiology of persistent digestive disorders (≥ 14 days) is poorly understood in low-resource settings and conventional diagnostic approaches lack accuracy. In this multi-country study, we compared multiplex real-time PCR for enteric bacterial, parasitic and viral pathogens in stool samples from symptomatic patients and matched asymptomatic controls in Côte d’Ivoire, Mali and Nepal. Among 1826 stool samples, the prevalence of most pathogens was highest in Mali, being up to threefold higher than in Côte d’Ivoire and up to tenfold higher than in Nepal. In all settings, the most prevalent bacteria were EAEC (13.0–39.9%) and Campylobacter spp. (3.9–35.3%). Giardia intestinalis was the predominant intestinal protozoon (2.9–20.5%), and adenovirus 40/41 was the most frequently observed viral pathogen (6.3–25.1%). Significantly different prevalences between symptomatic and asymptomatic individuals were observed for Campylobacter, EIEC and ETEC in the two African sites, and for norovirus in Nepal. Multiple species pathogen infection was common in Côte d’Ivoire and Mali, but rarely found in Nepal. We observed that molecular testing detected multiple enteric pathogens and showed low discriminatory accuracy to distinguish between symptomatic and asymptomatic individuals. Yet, multiplex PCR allowed for direct comparison between different countries and revealed considerable setting-specificity.

Case Report: A rare infection of multidrug-resistant Aeromonas caviae in a pediatric case with acute lymphoblastic leukemia and review of the literature

Aeromonas caviae infection of the bloodstream and intestine is a rare and severe opportunistic infection in immunocompromised people. In Southwest China, we first reported a case of bloodstream and intestinal infection with multidrug-resistant (MDR) Aeromonas caviae in a 4-year-old child with T-cell acute lymphoblastic leukemia. Blood and stool cultures were used to identify the infection. The selection of antibiotics was based on clinical expertise and medication sensitivity tests. We used linezolid, levofloxacin, and polymyxin B to treat the patient aggressively. Aeromonas caviae infection is uncommon in juvenile acute lymphoblastic leukemia. Doctors should be aware of the likelihood of opportunistic infection during the post-chemotherapy bone marrow suppression period. We further conducted a review of the literature and performed a detailed analysis of Aeromonas infection in pediatric leukemia. It is becoming increasingly apparent that antibiotic is abused domestically and abroad, resulting in the sharp increase of MDR bacteria. In general, most of the Aeromonas isolates are susceptible to third- or fourth-generation cephalosporins, aminoglycosides, quinolones, and carbapenem, but drug-resistant strains are being reported increasingly. We summarized the drug resistance rate of Aeromonas caviae and Aeromonas hydrophila in China in the last 10 years. Early recognition and effective treatment will improve prognosis and reduce mortality.

The Impacts of the Low-FODMAP Diet on the Management of Irritable Bowel Syndrome Symptoms: Possibilities, Feasibility, and Alternatives

Irritable bowel syndrome (IBS) is a common gastrointestinal (GI) condition characterized by a myriad of GI symptoms (e.g., diarrhea, nutrient malabsorption) that vary in severity. Due to its complex and individual nature, there is no known cure for IBS, but many diets have been assessed for their viability in managing symptoms. The low-FODMAP diet has recently been investigated for its potential benefits for IBS patients. FODMAPs, or Fermentable Oligosaccharides, Disaccharides, Monosaccharides, and Polyols are short- and medium-chain carbohydrates that are poorly absorbed in the small intestine and are prone to absorb water and ferment in the colon. Consumption of these FODMAPs correlates with increasing severity of IBS symptoms. The purpose of this randomized controlled trial systematic review was to compile various evidence-based dietary recommendations for the low-FODMAP diet for adult patients suffering from IBS from randomized controlled and clinical trials. The possibilities, feasibilities, and potential alternatives are proposed to determine if there is sufficient evidence to recommend the low-FODMAP diet for this patient population. Over 1,000 studies were screened and 6 were reviewed to determine the benefits of this diet. It was found that the low-FODMAP diet has demonstrated efficacy in delivering IBS symptom relief by changing patients’ metabolomes, microbiomes, and physiology. Specifically, this diet showed a greater overall decrease in abdominal pain, bloating, stool consistency, frequency, and urgency. Greater diversity of actinobacteria was found in the stool samples of these patients. Nociceptive neurons were also shown to be less sensitive and GI gas production was also markedly decreased. Despite these benefits, the low-FODMAP diet is more recommended for acute flare-ups due to its difficult adherence and attrition. Instead, we conclude that the balanced Mediterranean diet contains aspects of the traditional Mediterranean and low-FODMAP diet that can relieve IBS symptoms for daily life, while the low-FODMAP diet can be an effective treatment to ameliorate conditions when severe symptoms are experienced.

The gut microbiome, resistome, and mycobiome in preterm newborn infants and mouse pups: lack of lasting effects by antimicrobial therapy or probiotic prophylaxis

BackgroundEnhancing our understanding of the underlying influences of medical interventions on the microbiome, resistome and mycobiome of preterm born infants holds significant potential for advancing infection prevention and treatment strategies. We conducted a prospective quasi-intervention study to better understand how antibiotics, and probiotics, and other medical factors influence the gut development of preterm infants. A controlled neonatal mice model was conducted in parallel, designed to closely reflect and predict exposures. Preterm infants and neonatal mice were stratified into four groups: antibiotics only, probiotics only, antibiotics followed by probiotics, and none of these interventions. Stool samples from both preterm infants and neonatal mice were collected at varying time points and analyzed by 16 S rRNA amplicon sequencing, ITS amplicon sequencing and whole genome shotgun sequencing.ResultsThe human infant microbiomes showed an unexpectedly high degree of heterogeneity. Little impact from medical exposure (antibiotics/probiotics) was observed on the strain patterns, however, Bifidobacterium bifidum was found more abundant after exposure to probiotics, regardless of prior antibiotic administration. Twenty-seven antibiotic resistant genes were identified in the resistome. High intra-variability was evident within the different treatment groups. Lastly, we found significant effects of antibiotics and probiotics on the mycobiome but not on the microbiome and resistome of preterm infants.ConclusionsAlthough our analyses showed transient effects, these results provide positive motivation to continue the research on the effects of medical interventions on the microbiome, resistome and mycobiome of preterm infants.

Collider Bias Assessment in Colombian Indigenous Wiwa and Kogui Populations with Chronic Gastroenteric Disorder of Likely Infectious Etiology Suggests Complex Microbial Interactions Rather Than Clear Assignments of Etiological Relevance

Multiple microbial detections in stool samples of indigenous individuals suffering from chronic gastroenteric disorder of a likely infectious origin, characterized by recurring diarrhea of variable intensity, in the rural north-east of Colombia are common findings, making the assignment of etiological relevance to individual pathogens challenging. In a population of 773 indigenous people from either the tribe Wiwa or Kogui, collider bias analysis was conducted comprising 32 assessed microorganisms including 10 bacteria (Aeromonas spp., Campylobacter spp., enteroaggregative Escherichia coli (EAEC), enteropathogenic Escherichia coli (EPEC), enterotoxigenic Escherichia coli (ETEC), Salmonella spp., Shiga toxin-producing Escherichia coli (STEC), Shigella spp./enteroinvasive Escherichia coli (EIEC), Tropheryma whipplei and Yersinia spp.), 11 protozoa (Blastocystis spp., Cryptosporidium spp., Cyclospora spp., Dientamoeba fragilis, Entamoeba coli, Entamoeba bangladeshi/dispar/histolytica/moshkovskii complex, Entamoeba histolytica, Endolimax nana, Giardia duodenalis, Iodamoeba buetschlii and Pentatrichomonas hominis), 8 helminths (Ascaris spp., Enterobius vermicularis, Hymenolepis spp., Necator americanus, Schistosoma spp., Strongyloides spp., Taenia spp. and Trichuris spp.), microsporidia (Encephalocytozoon spp.) and fungal elements (microscopically observed conidia and pseudoconidia). The main results indicated that negative associations potentially pointing towards collider bias were infrequent events (n = 14), while positive associations indicating increased likelihood of co-occurrence of microorganisms quantitatively dominated (n = 88). Microorganisms showing the most frequent negative associations were EPEC (n = 6) and Blastocystis spp. (n = 3), while positive associations were most common for Trichuris spp. (n = 16), Dientamoeba fragilis (n = 15), Shigella spp./EIEC (n = 12), Ascaris spp. (n = 11) and Blastocystis spp. (n = 10). Of note, positive associations quantitively dominated for Blastocystis spp. In conclusion, collider bias assessment did not allow clear-cut assignment of etiological relevance for detected enteric microorganisms within the assessed Colombian indigenous population. Instead, the results suggested complex microbial interactions with potential summative effects. Future studies applying alternative biostatistical approaches should be considered to further delineate respective interactions.

Epidemiological Characteristics of Shiga Toxin-Producing Escherichia coli Responsible for Infections in the Polish Pediatric Population.

Shiga toxin-producing Escherichia coli (STEC) are zoonotic pathogens causing hemorrhagic colitis and hemolytic uremic syndrome (HUS) in children and the elderly. Stool samples were collected from 180 children hospitalized in five pediatric centers in Poland in 2018-2022. Direct stx1/stx2 gene detection by PCR in feces and E. coli isolates was performed. Antibiotic susceptibility was tested according to EUCAST v.12. Randomly selected isolates were serotyped with O157 antiserum and genotyped by pulsed-field gel electrophoresis (PFGE). A total of 44 E. coli isolates were confirmed as STEC by PCR. Among them, 84.4% were positive for stx2, and equally 6,8% for only stx1 and both stx1 and stx2 genes. The stx1 gene was also found in one Citrobacter freundii isolate. E. coli serotype O157 was present in 97.6% of the isolates. STEC infections most often occurred between June-October with a peak in July and August (51%). The highest, 77.8% of STEC isolates were found in the 1-5 years old group. No extended-spectrum β-lactamases (ESBL) were found. Resistance only to amoxicillin/clavulanic acid (24.4%), piperacillin/tazobactam (3%), cefotaxime (6%), gentamicin (6%), ciprofloxacin (3%), azithromycin (3%), trimethoprim/sulfamethoxazole (24,2%) was detected. PFGE analysis showed 18 PFGE types with no clonal distribution. Eight isolates with A, B, and C PFGE types showed genetic relatedness in the type with no detection of transmission way of distribution. STEC strains pose a serious threat to human health, therefore demographic and epidemiological characteristics are crucial for their surveillance.

Prevalence of Intestinal Helminths Among Mogadishu Residents Attending Somali Sudanese Specialized Hospital, Mogadishu, Somalia

Intestinal helminths infection remains a public health concern in the developing countries. Over 1.5 billion people on a global scale are infected with one or more intestinal helminths. Sub - Saharan Africa is considered among the most affected areas with helminths infections. These infection rates are linked with factors such as socioeconomic status, personal hygiene, overcrowding and poor waste management. This study aimed to investigate the frequency of intestinal helminths infections among Mogadishu residents attending Somali Sudanese specialized hospital. The presented study was designed as a descriptive cross - sectional hospital - based study conducted at Somali Sudanese specialized hospital at Mogadishu, Somalia, during the period from November 2022 to April 2023. Stool samples were collected from 1865 Mogadishu residents attending Somali Sudanese Specialized Hospital. Helminths infections were confirmed by identification of the diagnostic stage of the parasite under light microscope. The study findings indicated that the prevalence of intestinal helminths was 2.0%. The distribution of the parasitic infections was 0.7%, 0.6%, 0.4%, 0.3% (Trichuris trichiura, Enterobius vermicularis, Hymenolepis nana, and Ascaris lumbricoides) respectively. This result showed that the most common infection is caused by Trichuris trichiura followed by Enterobius vermicularis with Ascaris lumbricoides being the least common. The high prevalence of intestinal helminths infections among Mogadishu residents highlights a poor personal hygiene.

Exploring biorepository donation patterns, experiences, and recommendations: a mixed-methods study among Appalachian adults enrolled in a sugary drink reduction program

BackgroundUnder-represented subgroups in biomarker research linked to behavioral health trials may impact the promise of precision health. This mixed methods study examines biorepository donations across an Appalachian sample enrolled in a sugary drink reduction intervention trial.MethodsParticipants enrolled in the behavioral trial were asked to join an optional biomarker study and were tracked for enrollment and biospecimen returns (stool and/or buccal sample). At 6 months, participants completed a summative interview on decision-making process, experiences collecting samples, and recommendations to encourage biospecimen donation. Return rates were analyzed across demographics (i.e., age, gender, race, education, income, health literacy status, and rurality status) using chi-squares. Qualitative data were content coded with differences compared by biomarker study enrollment and donation choices.ResultsOf the 249 invited participants, 171 (61%) enrolled, and 63% (n = 157) returned buccal samples and 49% (n = 122) returned stool samples. Metro residing participants were significantly more likely (56%) to return stool samples compared to non-metro (39%) counterparts [x2(1) = 6.61; p = 0.01]. Buccal sample return had a similar trend, 67 and 57%, respectively for metro vs. non-metro [x2(1) = 2.84; p = 0.09]. An additional trend indicated that older (≥40 years) participants were more likely (55%) to donate stool samples than younger (43%) participants [x2(1) = 3.39; p = 0.07]. No other demographics were significantly associated with biospecimen return. Qualitative data indicated that societal (66–81%) and personal (41–51%) benefits were the most reported reasons for deciding to donate one or both samples, whereas mistrust (3–11%) and negative perceptions of the collection process (44–71%) were cited the most by those who declined one or both samples. Clear instructions (60%) and simple collection kits (73%) were donation facilitators while challenges included difficult stool collection kits (16%) and inconveniently located FedEx centers (16%). Recommendations to encourage future biorepository donation were to clarify benefits to science and others (58%), provide commensurate incentives (25%), explain purpose (19%) and privacy protections (20%), and assure ease in sample collection (19%).ConclusionStudy findings suggest the need for biomarker research awareness campaigns. Researchers planning for future biomarker studies in medically underserved regions, like Appalachia, may be able to apply findings to optimize enrollment.

Seroprevalence of Helicobacter pylori IgG Antibodies and Associated Risk Factors among Asymptomatic Vegetable Sellers in Uyo, South Nigeria

Background: Vegetables are essential part of people’s diet and are sometimes consumed raw and often without heat treatment or thorough washing, hence some studies suggested it serves as vehicle for the transmission of pathogenic microorganisms like Helicobacter pylori. The source of contamination has also been traced to sellers of vegetables who are involved in unhygienic practices which contribute to the dissemination of pathogenic microorganisms. Therefore the aim of this study was to provide information on the seroprevalence of H. pylori IgG antibodies and associated risk factors among asymptomatic vegetable sellers in Uyo, Akwa Ibom State, Nigeria. Materials and Methods: Blood and stool samples from 142 participants were included in the study. Blood samples were analyzed using the rapid diagnostic method while the stool samples were analyzed using Enzyme linked immunosorbent Assays (ELISA). Results: A total of 85 (59.9%) were positive by rapid diagnostic test (RDT) while 53 (37.3%) were positive by stool ELISA test. Risk factors like age, marital status, residency family size, years in vegetable business, source of vegetable, wash before stalling etc were all statistically associated with (p&lt;0.05) with H. pylori prevalence. Conclusion: This study revealed that unhygienic behaviors by vegetable sellers are associated with high risk of H. pylori infection to vegetable consumers.

The diversity of the microbiome impacts chronic lymphocytic leukemia development in mice and humans.

The gut microbiota play a critical role in maintaining a healthy human body and their dysregulation is associated with various diseases. In this study, we investigated the influence of the gut microbiome diversity on chronic lymphocytic leukemia (CLL) development. Stool sample analysis of 59 CLL patients revealed individual and heterogeneous microbiome compositions, but allowed for grouping of patients according to their microbiome diversity. Interestingly, CLL patients with a lower microbiome diversity and an enrichment of bacteria linked to poor health suffered from a more advanced or aggressive form of CLL. In the Eμ-TCL1 mouse model of CLL, we observed a faster course of disease when mice were housed in high hygiene conditions. Shotgun DNA sequencing of fecal samples showed that this was associated with a lower microbiome diversity which was dominated by Mucispirillum and Parabacteroides genera in comparison to mice kept under lower hygiene conditions. In conclusion, we applied taxonomic microbiome analyses to demonstrate a link between the gut microbiome diversity and the clinical course of CLL in humans, as well as the development of CLL in mice. Our novel data serve as a basis for further investigations to decipher the pathological and mechanistic role of intestinal microbiota in CLL development.

Retrospective Genotyping of Enteroviruses Using a Diagnostic Nanopore Sequencing Workflow

Enteroviruses are among the most common viruses pathogenic to humans. They are associated with various forms of disease, ranging from mild respiratory illness to severe neurological diseases. In recent years, an increasing number of isolated cases of children developing meningitis or encephalitis as a result of enterovirus infection have been reported, as well as discrete enterovirus D68 outbreaks in North America in 2014 and 2016. We developed an assay to rapidly genotype enteroviruses by sequencing a region within the VP1 gene using nanopore Flongles. We retrospectively analyzed enterovirus-/rhinovirus-positive clinical samples from the Zurich, Switzerland area mainly collected during two seasons in 2019/2020 and 2021/2022. Respiratory, cerebrospinal fluid, and stool samples were analyzed. Whole-genome sequencing was performed on samples with ambiguous genotyping results and enterovirus D68-positive samples. Out of 255 isolates, a total of 95 different genotypes were found. A difference in the prevalence of enterovirus and rhinovirus infections was observed for both sample type and age group. In particular, children aged 0–4 years showed a higher frequency of enterovirus infections. Comparing the respiratory seasons, a higher prevalence was found, especially for enterovirus A and rhinovirus A after the SARS-CoV-2 pandemic. The enterovirus genotyping workflow provides a rapid diagnostic tool for individual analysis and continuous enterovirus surveillance.

Differential Composition and Structure of the Microbiota from Active and Inactive Stages of HLA-B27-associated Uveitis by Paired Fecal Metagenomes

ABSTRACT Purpose To compare the diversities and abundances of bacterial taxa in the microbiome of patients with HLA B27-positive acute anterior uveitis (AAU) in the active and inactive phases. Methods An observational descriptive prospective and comparative study was conducted in ten HLA-B27-positive AAU patients (44.6 ± 13.4 years). The microbiome of the stool samples obtained in the active and inactive stages was analyzed by sequencing the V3 region of the 16S rRNA gene. Results The differences in the bacteria profile between active and inactive stages in each individual were confirmed (p < 0.0001). Ten OTUs were found exclusively in the active phase of 90% of the individuals, suggesting a proinflammatory association. Blautia OUT_4 and Faecalibacterium OUT_2 abundances showed a direct relationship between abundance and severity of ocular inflammation. Two OTUs were exclusive of the inactive stage, suggesting an anti-inflammatory role. Conclusion The metagenomic profile of the fecal microbiota differs in the acute phase of the AAU compared to when the inflammation subsides, despite being the same individual and a short time-lapse. AAU is a fertile field for studying the connection between subtle rapid changes in microbiota and their systemic consequences.

Occurrence of Free-Living Amoebae in Non-Human Primate Gut

The gut microbiome reflects health and predicts possible disease in hosts. A holistic view of this community is needed, focusing on identifying species and dissecting how species interact with their host and each other, regardless of whether their presence is beneficial, inconsequential, or detrimental. The distribution of gut-associated eukaryotes within and across non-human primates is likely driven by host behavior and ecology. To ascertain the existence of free-living amoebae (FLA) in the gut of wild and captive non-human primates, 101 stool samples were collected and submitted to culture-dependent microscopy examination and DNA sequencing. Free-living amoebae were detected in 45.4% (46/101) of fecal samples analyzed, and their morphological characteristics matched those of Acanthamoeba spp., Vermamoeba spp., heterolobosean amoeboflagellates and fan-shaped amoebae of the family Vannellidae. Sequence analysis of the PCR products revealed that the suspected amoebae are highly homologous (99% identity and 100% query coverage) with Acanthamoeba T4 genotype and Vermamoeba vermiformis amoebae. The results showed a great diversity of amoebae in the non-human primate’s microbiome, which may pose a potential risk to the health of NHPs. To our knowledge, this is the first report of free-living amoebae in non-human primates that are naturally infected. However, it is unknown whether gut-borne amoebae exploit a viable ecological niche or are simply transient residents in the gut.