Bacteroides Microbial Source Tracking Markers Perform Poorly in Predicting Enterobacteriaceae and Enteric Pathogen Contamination of Cow Milk Products and Milk-Containing Infant Food

Consumption of microbiologically contaminated food is one of the leading causes of diarrheal diseases. Understanding the source of enteric pathogens in food is important to guide effective interventions. Enterobacteriaceae bacterial assays typically used to assess food safety do not shed light on the source. Source-specific Bacteroides microbial source tracking (MST) markers have been proposed as alternative indicators for water fecal contamination assessment but have not been evaluated as an alternative fecal indicator in animal-derived foods. This study tested various milk products collected from vendors in urban Kenyan communities and infant foods made with the milk (n = 394 pairs) using conventional culture methods and TaqMan qPCR for enteric pathogens and human and bovine-sourced MST markers. Detection profiles of various enteric pathogens and Bacteroides MST markers in milk products differed from that of milk-containing infant foods. MST markers were more frequently detected in infant food prepared by caregivers, indicating recent contamination events were more likely to occur during food preparation at home. However, Bacteroides MST markers had lower sensitivity in detecting enteric pathogens in food than traditional Enterobacteriaceae indicators. Bacteroides MST markers tested in this study were not associated with the detection of culturable Salmonella enterica and Shigella sonnei in milk products or milk-containing infant food. The findings show that while Bacteroides MST markers could provide valuable information about how foods become contaminated, they may not be suitable for predicting the origin of the enteric pathogen contamination sources.

Environmental and behavioral exposure pathways associated with diarrhea and enteric pathogen detection in twenty-six week old urban Kenyan infants: a cross-sectional study

The prevalence of enteric pathogen detection in children in low-income countries climbs rapidly between birth and 6 months of age. Few studies have tested whether improved household environmental and behavioral hygiene conditions protects infants from exposure to enteric pathogens spread via unhygienic human and animal sanitation conditions, especially during this early window of infancy. This cross-sectional study utilized enrollment survey data among households with 6 month old infants in Kisumu, Kenya participating in the Safe Start cluster-randomized controlled trial to estimate associations between household water access and treatment, animal vectors, sanitation access, hand washing practices, supplemental feeding, and flooring, with the outcomes of caregiver-reported 7-day diarrhea prevalence and sum count of different enteric viruses, bacteria, and parasites pathogens in infant stool. Then, we tested whether household environmental hygiene and behavioral practices moderated associations between infant exposure outcomes and latrine access and domestic animal co-habitation. We found that reported handwashing after handling animals and before eating were strongly associated with lower risk of caregiver-reported diarrhea, while owning and co-habitating with animals (versus no animals), living in a household with vinyl covered dirt floors (versus finished floors), and feeding infants cow milk (versus no milk) were strongly associated with pathogen detection in infants. Caregiver handwashing after child or self-defecation moderated the relationship between shared sanitation (vs private) sanitation access and infant exposure to pathogens such that handwashing had the greatest benefit for preventing pathogen exposure of infants in households with private latrines. In the absence of handwashing, access to private sanitation posed no benefits over shared latrines for protecting infants from exposure. Our evidence highlights eliminating animal co-habitation, improving flooring, improving post-defecation and food-related handwashing, and improving safety of cow milk sources and/or safe household storage of milk as interventions to prevent enteric pathogen exposure of infants less than 6 months age.

Enteric pathogen among children under five years old with diarrheal diseases in Indonesia

Diarrheal diseases are the second cause of the high morbidity and mortality in children under five years old. According to the Basic Health Survey 2018 conducted by the Ministry of Health, the prevalence of diarrheal diseases among children under five years old that were diagnosed by healthcare workers was 11.0%. The aim of this study was to describe the enteric pathogen isolated from children with diarrhea. The study was conducted in five cities in Indonesia: Jakarta, Serang, Denpasar, Makassar, and Mataram. The Inclusion criteria were children aged one month to five years old, with diarrhea that was diagnosed by a healthcare worker. The rectal swabs were sent to the Centre for Research and Development for Biomedical and Basic Health Technology, National Institute of Health Research and Development, Ministry of Health in Jakarta. Virus and Enterotoxigenic Escherichia coli (ETEC) identification by using multiplex PCR from Seegene, meanwhile bacteria identified by conventional method. As many as 2626 children under five years old participated in this study. The highest viral pathogen that causes diarrhea is viral 1.807 (68,81%) and 486 (18,56%). The virus etiology was Rotavirus 982 (54,34%) cases, followed by Adenovirus 916 (50.69) cases, Norovirus II 444 (24,57%) cases, meanwhile the bacteria pathogen were Enterotoxigenic Escherichia coli detected in 262 (9,98%) followed by Campylobacter jejuni and Shigella spp. This study described Rotavirus is the prevalence etiology of diarrhea among children under five years old followed by Adenovirus and Norovirus, some other cases reported the cause of diarrhea were bacteria ETEC E. coli followed Campylobacter jejuni, Shigella spp, etc.

Effect of scheduled antimicrobial and nicotinamide treatment on linear growth in children in rural Tanzania: A factorial randomized, double-blind, placebo-controlled trial

Background Stunting among children in low-resource settings is associated with enteric pathogen carriage and micronutrient deficiencies. Our goal was to test whether administration of scheduled antimicrobials and daily nicotinamide improved linear growth in a region with a high prevalence of stunting and enteric pathogen carriage. Methods and findings We performed a randomized, 2 × 2 factorial, double-blind, placebo-controlled trial in the area around Haydom, Tanzania. Mother–child dyads were enrolled by age 14 days and followed with monthly home visits and every 3-month anthropometry assessments through 18 months. Those randomized to the antimicrobial arm received 2 medications (versus corresponding placebos): azithromycin (single dose of 20 mg/kg) at months 6, 9, 12, and 15 and nitazoxanide (3-day course of 100 mg twice daily) at months 12 and 15. Those randomized to nicotinamide arm received daily nicotinamide to the mother (250 mg pills months 0 to 6) and to the child (100 mg sachets months 6 to 18). Primary outcome was length-for-age z-score (LAZ) at 18 months in the modified intention-to-treat group. Between September 5, 2017 and August 31, 2018, 1,188 children were randomized, of whom 1,084 (n = 277 placebo/placebo, 273 antimicrobial/placebo, 274 placebo/nicotinamide, and 260 antimicrobial/nicotinamide) were included in the modified intention-to-treat analysis. The study was suspended for a 3-month period by the Tanzanian National Institute for Medical Research (NIMR) because of concerns related to the timing of laboratory testing and the total number of serious adverse events (SAEs); this resulted in some participants receiving their final study assessment late. There was a high prevalence of stunting overall (533/1,084, 49.2%). Mean 18-month LAZ did not differ between groups for either intervention (mean LAZ with 95% confidence interval [CI]: antimicrobial: −2.05 CI −2.13, −1.96, placebo: −2.05 CI −2.14, −1.97; mean difference: 0.01 CI −0.13, 0.11, p = 0.91; nicotinamide: −2.06 CI −2.13, −1.95, placebo: −2.04 CI −2.14, −1.98, mean difference 0.03 CI −0.15, 0.09, p = 0.66). There was no difference in LAZ for either intervention after adjusting for possible confounders (baseline LAZ, age in days at 18-month measurement, ward, hospital birth, birth month, years of maternal education, socioeconomic status (SES) quartile category, sex, whether the mother was a member of the Datoga tribe, and mother’s height). Adverse events (AEs) and SAEs were overall similar between treatment groups for both the nicotinamide and antimicrobial interventions. Key limitations include the absence of laboratory measures of pathogen carriage and nicotinamide metabolism to provide context for the negative findings. Conclusions In this study, we observed that neither scheduled administration of azithromycin and nitazoxanide nor daily provision of nicotinamide was associated with improved growth in this resource-poor setting with a high force of enteric infections. Further research remains critical to identify interventions toward improved early childhood growth in challenging conditions. Trial registration ClinicalTrials.gov NCT03268902.

Deprivation of dietary fiber in specific-pathogen-free mice promotes susceptibility to the intestinal mucosal pathogen Citrobacter rodentium

The change of dietary habits in Western societies, including reduced consumption of fiber, is linked to alterations in gut microbial ecology. Nevertheless, mechanistic connections between diet-induced microbiota changes that affect colonization resistance and enteric pathogen susceptibility are still emerging. We sought to investigate how a diet devoid of soluble plant fibers impacts the structure and function of a conventional gut microbiota in specific-pathogen-free (SPF) mice and how such changes alter susceptibility to a rodent enteric pathogen. We show that absence of dietary fiber intake leads to shifts in the abundances of specific taxa, microbiome-mediated erosion of the colonic mucus barrier, a reduction of intestinal barrier-promoting short-chain fatty acids, and increases in markers of mucosal barrier integrity disruption. Importantly, our results highlight that these low fiber diet-induced changes in the gut microbial ecology collectively contribute to a lethal colitis by the mucosal pathogen Citrobacter rodentium, which is used as a mouse model for enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively). Our study indicates that modern, low-fiber Western diets might make individuals more prone to infection by enteric pathogens via the disruption of mucosal barrier integrity by diet-driven changes in the gut microbiota, illustrating possible implications for EPEC and EHEC infections.