Feasibility of an At-Home Adult Stool Specimen Collection Method in Rural Cambodia

The human microbiome has received significant attention over the past decade regarding its potential impact on health. Epidemiological and intervention studies often rely on at-home stool collection methods designed for high-resource settings, such as access to an improved toilet with a modern toilet seat. However, this is not always appropriate or applicable to low-resource settings. Therefore, the design of a user-friendly stool collection kit for low-resource rural settings is needed. We describe the development, assembly, and user experience of a simple and low-cost at-home stool collection kit for women living in rural Cambodia as part of a randomized controlled trial in 2020. Participants were provided with the stool collection kit and detailed verbal instruction. Enrolled women (n = 480) provided two stool specimens (at the start of the trial and after 12 weeks) at their home and brought them to the health centre that morning in a sterile collection container. User specimen collection compliance was high, with 90% (n = 434) of women providing a stool specimen at the end of the trial (after 12 weeks). This feasible and straightforward method has strong potential for similar or adapted use among adults residing in other rural or low-resource contexts.

ROLE OF ABSOLUTE EOSINOPHIL COUNT AS A MODALITY OF INVESTIGATION IN SUSPECTED CASES OF HELMINTHIC INFESTATION IN CHILDREN AGED BETWEEN 5 TO 15 YEARS WITH COMPARISON TO STOOL ROUTINE AND MICROSCOPY

Introduction: Eosinophilia is a well-known host immune response in helminthic infestation. This study was carried out to investigate whether a correlation exists between absolute eosinophil count in children with stool specimen positive for helminthic infestation. Aim And Objective: To determine average raised absolute eosinophil count in children who were having various helminthic infestation with stool specimen positive. Materials And Methods: A hospital based cross sectional study done at Navodaya medical college hospital and research centre during 2019 October to 2020 September for a period of 12 months. Both blood and stool samples were obtained from a total of 69 suspected children after examined, categorised into 2 groups. Among them 24 cases were positive of helminthic infestation and 45 cases were negative. T-test and descriptive statistics were used to analyse the data. Results: The study revealed that children infested with helminths found to have higher absolute eosinophil count (mean 4644/µl (95%CI; 3474-4212) than not infested (mean 801/µl (95%CI; 616-945) with p value <0.001. In this helminth's family; nematodes, trematodes, cestodes showed mean values of absolute eosinophil count were 5206/µl (4806-7151/µl), 4930/µl (4500-5382/µl) and 2179/µl (2500-1669/µl) respectively. Conclusions: Raised absolute eosinophil count has been observed in stool specimen positive cases of helminths, especially more in nematodes. Therefore, should be considered for a single dose of albendazole.

Blautia intestinalis sp. nov., isolated from human feces

A strictly anaerobic bacterial strain (27-44T) was isolated from a stool specimen from an autistic child collected in PR China. The strain was Gram-stain-positive, non-motile, non-pigmented, non-spore-forming, and cells were oval to rod-shaped. Strain 27-44T grew at 20–40 °C (optimal at 37 °C) and at pH 6.0–10 (optimal at 6.0–8.0). The major polar lipids were one phospholipid, two glycolipids, two aminophospholipids and one unidentified lipid. The major cellular fatty acids of strain 27-44T were C16 : 0 and C17 : 0 2-OH. The end product of glucose fermentation was mainly butyric acid. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain 27-44T was a member of the genus Blautia and phylogenetically closely related to Blautia obeum ATCC 29174T (with 97.8 % seque nce similarity). The genome of strain 27-44T was 3.5 Mbp with a DNA G+C content of 42.36 mol%. A total of 3436 genes were predicted and, of these, 3133 genes were annotated by KEGG. On the basis of phenotypic, chemotaxonomic and phylogenetic comparisons, strain 27-44T represents a novel species within the genus Blautia , for which the name Blautia intestinalis sp. nov. is proposed. The type strain is 27-44T= CGMCC 1.5285T=NBRC 113774T.

Characterization of enteric disease in children using a low-cost specimen preservation method

BACKGROUND: Diarrhea is a leading cause of death in children under five. Molecular methods exist for the rapid detection of enteric pathogens; however, the logistical costs of storing stool specimens limit applicability. We sought to demonstrate that dried filter paper specimen preservation can identify diarrheal diseases causing significant morbidity among children in resource constrained countries. METHODS: A sub-study was nested into cholera surveillance in Cameroon. Enrollment criteria included: enrollment between 8/1/16 - 10/1/18; age < 18 years; a stool specimen; ≥ three loose stools within 24hours with the presence of dehydration and/or blood. 7227 persons were enrolled, for which 2746 met enrollment criteria and 337 were included in this analysis using the enteric TaqMan Array Card. Bacterial pathogens were compared to severity of diarrhea, age and sex, among other variables. RESULTS: 107 were ETEC positive of which: 40.2% (N=43) LT-STh, 19.6% (N=21) LT-STp; and 49.5% (53) LT-only. Major CFs were present in 43.9% of ETEC-positives. 96 were positive for Shigella, of which 14 (14.6%) reported dysentery. Model-derived quantitative cutoffs identified 116 (34.4%) with one highly diarrhea-associated pathogen and 16 (4.7%) with ≥ two. Shigellae and rotavirus were most strongly associated with diarrhea in children with mixed infections. CONCLUSION: Dried filter paper preserved specimens eliminate the need for frozen stool specimens and will facilitate enteric surveillance and contribute to the understanding of disease burden, which is needed to guide vaccine development and introduction. This study confirms Rotavirus, Shigella and ETEC as major contributors to pediatric diarrheal disease in two regions of Cameroon.

Discovery and Full Genome Characterization of SARS-CoV-2 in Stool Specimen from a Recovered Patient, China

SARS-CoV-2 was found in a recovered patient’s stool specimen by combining quantitative reverse transcription PCR (qRT-PCR) and genome sequencing. The patient was virus positive in stool specimens for at least an additional 15 days after he was recovered, whereas respiratory tract specimens were negative. The discovery of the complete genome of SARS-CoV-2 in the stool sample of the recovered patient demonstrates a cautionary warning that the potential mode of the virus transmission cannot be excluded through the fecal-oral route after viral clearance in the respiratory tract.

Intra-specimen and day-to-day variations of Fasciola egg counts in human stools

Intra-specimen and day-to-day variations of Fasciola egg counts in stools were investigated for 16 cases of established fascioliasis. For each case six Kato slides from a single stool sample were examined daily for 5 consecutive days. The results indicated the presence of significant intra-specimen variations in more than one-third of the examined series, while the inter-specimen variation was almost negligible. The sensitivity of the Kato-Katz test for diagnosing Fasciola infection with three Kato slides from the same specimen or on different days ranged from 96.0%-99.1%. The examination of three Kato smears from a single stool specimen, which is more feasible in field studies, would give an accurate diagnosis of fascioliasis. Used as such, the Kato-Katz technique is highly sensitive in the diagnosis of fascioliasis

Closed Genome Sequence of Clostridium botulinum Strain IBCA10-7060 Type Bh

Clostridium botulinum strain IBCA10-7060 was isolated from a stool specimen from an infant botulism patient and is the only Clostridium botulinum strain known that produces botulinum toxin type H. We present here its 4.09-Mbp closed genome sequence.

Stool specimen for diagnosis of pulmonary tuberculosis in adults: protocol for a systematic review and meta-analysis

IntroductionTuberculosis (TB) continues to be a significant health burden, most commonly affecting the lungs and referred to as pulmonary TB (PTB). Diagnostic techniques of PTB primarily rely on expectorated sputum samples. However, the diagnostic yields are often hindered due to insufficient volume and quality of the sputum specimens. Moreover, some individuals are unable to provide sputum samples due to scanty sputum production or difficulty in coughing up and require an invasive procedure to obtain a respiratory sample, such as bronchoscopic or gastric aspiration. Thus, challenges in the acquisition of respiratory specimens warrant an alternate specimen. Therefore, this systematic review aims to evaluate the diagnostic accuracy of a stool specimen for the diagnosis of PTB in adults.Methods and analysisWe will search MEDLINE (Ovid), Embase (Ovid), Web of Science and Cochrane database from inception to April 2021 using a comprehensive search strategy. Two reviewers will independently perform screening, data extraction and quality assessment. The risk of bias assessment and applicability of results of eligible studies will be performed using the Quality of Diagnostic Accuracy Studies-2 tool. Bivariate random-effects models will be performed to calculate pooled sensitivity, specificity, positive likelihood ratio and negative likelihood ratio and diagnostic odds ratio along with 95% CI of stool specimen for each reported diagnostic method against any of the reference standard test (ie, mycobacterial culture or smear microscopy or Xpert assay using respiratory specimens). Heterogeneity between studies will be assessed by I2 statistics and Q statistic of the χ2 test.Ethics and disseminationThe results will be disseminated through publishing in a peer-reviewed medical journal and public presentations in relevant national and international conferences. As this is a systematic review of publicly available data, ethics approval is not required.PROSPERO registration numberCRD42021245203.

Water, sanitation, and hygiene conditions and prevalence of intestinal parasitosis among primary school children in Dessie City, Ethiopia

Background Intestinal parasitosis is a major public health problem that affects the health of primary school children in low- and middle-income countries where water, sanitation, and hygiene (WASH) conditions are deficient. Since there is a paucity of information on the prevalence and associated factors of this problem among primary school children in Dessie City in Ethiopia, this study was designed to address these gaps. Methods A school-based cross-sectional study was conducted among 407 stratified-sampled primary school children in five primary schools at Dessie City from April to June 2018. Data were collected using a pretested structured questionnaire, an observation checklist and laboratory analysis of stool samples. Stool specimen from each study participant was collected using clean, properly labeled and leak-proof stool cup. A portion of stool from each study participant collected sample was processed using saline wet mount technique and examined by microscope. The remaining specimens were preserved with 10% formalin and transported to Dessie Comprehensive Specialized Hospital laboratory to be processed by using formol-ether concentration technique. Then, slide smears were prepared from each processed stool specimen and finally, it was microscopically examined with 10x as well as 40x objectives for the presence or absence of intestinal parasites. Factors significantly associated with intestinal parasitosis were determined using binary logistic regression model at 95% CI (confidence interval). Thus, bivariate (COR [crude odds ratio]) and multivariable (AOR [adjusted odds ratio]) logistic regression analyses were carried out. From the multivariable analysis, variables having a p-value of less than 0.05 were declared as factors significantly associated with intestinal parasitosis among primary school children. Main findings The overall prevalence of intestinal parasitosis was found to be 16.0% (95% CI: 12.5–19.4%), of these, 50.8% were positive for protozoa, 32.2% for helminth infections and 16.9% for double co-infections. Entamoeba histolytica was the most prevalent parasite (29.2%), followed by Giardia lamblia (21.5%), Ascaris lumbricoides (18.5%), Hymenolepis nana (9.2%) and Enterobius vermicularis (4.6%). Prevalence rates were similar among government (16.3%) and private (15.7%) school children. Water consumption was less than 5 liters per capita per day in 4 of the 5 schools. Thirty-eight (9.3%) of primary school students reported that they practiced open defecation. About two-thirds (285, 70.0%) said they always washed their hands after defecation. Mother’s education (illiterate) (AOR = 3.3; 95% CI: 1.20–9.37), father’s education (illiterate) (AOR = 3.9; 95% CI: 1.40–10.82), fathers who could read and write (AOR = 3.3; 95% CI: 1.25–7.86), handwashing before meal (sometimes) (AOR = 2.2; 95% CI: 1.11–4.17) and poor knowledge of WASH (AOR = 9.3; 95% CI: 2.17–16.70) were statistically associated with presence of intestinal parasitic infections. Conclusion We concluded that the prevalence of intestinal parasitosis in the study area among Grades 4–8 primary school children had public health significance. Factors significantly associated with intestinal parasitosis among primary school children’s were illiterate mothers and fathers, irregular handwashing of children before meals, and poor knowledge of WASH. Health education to improve students’ WASH knowledge and mass deworming for parasites are recommended as preventive measures; and improvements to the quality of WASH facilities in primary schools are strongly recommended to support these measures.

Norovirus and Other Viral Causes of Medically Attended Acute Gastroenteritis Across the Age Spectrum: Results from the MAAGE Study in the United States

Background Acute gastroenteritis (AGE) causes a substantial burden in the United States, but its etiology frequently remains undetermined. Active surveillance within an integrated healthcare delivery system was used to estimate the prevalence and incidence of medically attended norovirus, rotavirus, sapovirus, and astrovirus. Methods Active surveillance was conducted among all enrolled members of Kaiser Permanente Northwest during July 2014 – June 2016. An age-stratified, representative sample of AGE-associated medical encounters were recruited to provide a stool specimen to be tested for norovirus, rotavirus, sapovirus, and astrovirus. Medically attended AGE (MAAGE) encounters for a patient occurring within 30 days were grouped into one episode, and all-cause MAAGE incidence was calculated. Pathogen- and healthcare setting-specific incidence estimates were calculated using age-stratified bootstrapping. Results The overall incidence of MAAGE was 40.6 episodes per 1000 person-years (PY), with most episodes requiring no more than outpatient care. Norovirus was the most frequently detected pathogen, with an incidence of 5.5 medically attended episodes per 1000 PY. Incidence of norovirus MAAGE was highest among children aged &lt;5 years (20.4 episodes per 1000 PY), followed by adults aged ≥65 years (4.5 episodes per 1000 PY). Other study pathogens showed similar patterns by age, but lower overall incidence (sapovirus: 2.4 per 1000 PY, astrovirus: 1.3 per 1000 PY, rotavirus: 0.5 per 1000 PY). Conclusions Viral enteropathogens, particularly norovirus, are an important contributor to MAAGE, especially among children &lt;5 years of age. The present findings underline the importance of judicious antibiotics use for pediatric AGE and suggest that an effective norovirus vaccine could substantially reduce MAAGE.