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Background: Limited prospective data are available on the acquisition of viral, bacterial and parasitic diarrhoeagenic agents by healthy individuals during travel.Methods: To determine the frequency of travel associated acquisition of 19 pathogens in 98 intercontinental travellers, qPCR was used to detect 8 viral pathogens, 6 bacterial enteric pathogens and 5 parasite species in faecal samples collected immediately before and after travel.Results: We found high pre-travel carriage rates of Blastocystis spp. and Dientamoeba fragilis of 32% and 19% respectively. Pre-travel prevalences of all other tested pathogens were below 3%. Blastocystis spp. (10%), Plesiomonas shigelloides (7%), D. fragilis (6%) and Shigella spp. (5%) were the most frequently acquired pathogens and acquisition of enteral viruses and hepatitis E virus in this relatively small group of travellers was rare or non-existent.Conclusions: Our findings suggest that the role of viruses as the cause of persisting traveller's diarrhoea is limited and bacterial pathogens are more likely as a cause of traveller's diarrhoea. The substantial proportion of travellers carrying Blastocystis spp. and D. fragilis before travel warrants cautious interpretation of positive samples in returning travellers with gastrointestinal complaints. (C) 2017 Elsevier Ltd. All rights reserved.

Summary Background International travel contributes to the dissemination of antimicrobial resistance. We investigated the acquisition of extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) during international travel, with a focus on predictive factors for acquisition, duration of colonisation, and probability of onward transmission. Methods Within the prospective, multicentre COMBAT study, 2001 Dutch travellers and 215 non-travelling household members were enrolled. Faecal samples and questionnaires on demographics, illnesses, and behaviour were collected before travel and immediately and 1, 3, 6, and 12 months after return. Samples were screened for the presence of ESBL-E. In post-travel samples, ESBL genes were sequenced and PCR with specific primers for plasmid-encoded β-lactamase enzymes TEM, SHV, and CTX-M group 1, 2, 8, 9, and 25 was used to confirm the presence of ESBL genes in follow-up samples. Multivariable regression analyses and mathematical modelling were used to identify predictors for acquisition and sustained carriage, and to determine household transmission rates. This study is registered with ClinicalTrials.gov, number NCT01676974. Findings 633 (34·3%) of 1847 travellers who were ESBL negative before travel and had available samples after return had acquired ESBL-E during international travel (95% CI 32·1–36·5), with the highest number of acquisitions being among those who travelled to southern Asia in 136 of 181 (75·1%, 95% CI 68·4–80·9). Important predictors for acquisition of ESBL-E were antibiotic use during travel (adjusted odds ratio 2·69, 95% CI 1·79–4·05), traveller's diarrhoea that persisted after return (2·31, 1·42–3·76), and pre-existing chronic bowel disease (2·10, 1·13–3·90). The median duration of colonisation after travel was 30 days (95% CI 29–33). 65 (11·3%) of 577 remained colonised at 12 months. CTX-M enzyme group 9 ESBLs were associated with a significantly increased risk of sustained carriage (median duration 75 days, 95% CI 48–102, p=0·0001). Onward transmission was found in 13 (7·7%) of 168 household members. The probability of transmitting ESBL-E to another household member was 12% (95% CI 5–18). Interpretation Acquisition and spread of ESBL-E during and after international travel was substantial and worrisome. Travellers to areas with a high risk of ESBL-E acquisition should be viewed as potential carriers of ESBL-E for up to 12 months after return. Funding Netherlands Organisation for Health Research and Development (ZonMw).

COMBAT consortium: et al., [Background]: We studied geographic distribution of diarrhoeagenic Escherichia coli virulence genes (DEC VGs) acquisition in travellers and investigated if they acquired highly virulent EAEC/STEC hybrid strains., [Methods]: From the prospective, multicentre COMBAT study among 2001 Dutch travellers, 491 travellers were selected based on travel destination to 7 subregions. Faecal samples taken directly before and after travel were screened for nine DEC VGs with real-time PCR. Incidence proportions and rates were calculated for each gene and subregion., [Results]: 479 travellers were analysed. 21.8% acquired aggR (EAEC), with highest acquisition rates in Northern and Western Africa and 15.3% acquired eae (STEC/EPEC) with highest rates in travellers to Western and Eastern Africa. ETEC (elt or est gene) was acquired by 4.2% of travellers and acquisition of est was associated with traveller's diarrhoea. Overall, the risk of acquiring DEC VGs was low in Southern Africa and South America. Although the combination of aggR (EAEC) and stx1/2 (STEC) was acquired by 3 travellers, these genes could not be detected together in a single E. coli strain., [Conclusions]: The risk of acquisition of DEC VGs strongly depends on the travel destination, with those travelling to Africa - except Southern Africa - having a higher risk., The COMBAT-study was funded by Netherlands Organisation for Health Research and Development (ZonMw, grant number 205200003). This work is a contribution to EU FP7 ANTIGONE (project number 278976).

SCOPE: Understanding biological functions of different free human milk oligosaccharides (HMOs) in shaping gastrointestinal tract microbiota during infancy is of great interest. We examined a link between HMOs in maternal milk and infant faecal microbiota composition and investigated the role of microbiota in degrading HMOs within the GI tract of healthy, breastfed, one-month old infants. METHODS AND RESULTS: Maternal breast milk and corresponding infant faeces originated from the KOALA Birth Cohort. HMOs were quantified in milk and infant faecal samples using PGC-UPLC-MS and HPAEC-PAD. Faecal microbiota composition was characterised using Illumina HiSeq amplicon 16S rRNA sequencing. The composition associated with gender, mode of delivery, and milk HMOs: Lacto-N-fucopentaose I and 2'-Fucosyllactose. Overall, Bifidobacterium, Bacteroides, Escherichia-Shigella and Parabacteroides were predominating genera. We detected three different patterns in the infant faecal microbiota structure. Gastrointestinal degradation of HMOs was strongly associated with faecal microbiota composition, and there was a link between utilisation of specific HMOs and abundance of various phylotypes (OTUs). CONCLUSIONS: HMOs in maternal milk are among important factors shaping GI tract microbiota composition in one-month old breastfed infants. Infant's ability to metabolise different HMOs strongly correlate with faecal microbiota composition, and with phylotypes within genera Bifidobacterium, Bacteroides and Lactobacillus. This article is protected by copyright. All rights reserved.

Figure S2. Gene count distribution in Dutch children. Black indicates all individuals, n = 281; red indicates lean children (BMI z-score

Figure S1. Compositional and functional comparison between Dutch children and adults. Relative abundance of major phyla in Dutch children (a) and adults (b). (c) Relative abundance of COG (clusters of orthologous groups) categories across each sample in Dutch children and adults. (PDF 298 kb)

Figure S4. Comparison of gut microbial functional potentials between enterotypes. (a) Differentially enrichment of KEGG modules between enterotypes. Dashed lines indicate a reporter score of 1.96, corresponding to 95% confidence in a normal distribution. (b) Heatmap showing that the relative abundance profiles of 8 selected KOs involved in key functions of metabolic pathways for carbohydrate metabolism (K00845, K01051 and K00873) and amino acid biosynthesis (K01738, K00928, K00058, K00651and K00765) distinguishes E3 from E1 and E2. (PDF 358 kb)

Figure S6. Correlations between continuous phenotypic parameters. (a) Spearman’s rank correlations between continuous phenotypic parameters in the entire cohort (n = 281). (b) Spearman’s correlations between continuous phenotypic parameters in E1 (n = 143). (c) Spearman’s rank correlations between continuous phenotypic parameters in E2 (n = 74). (d) Spearman’s rank correlations between continuous phenotypic parameters in E3 (n = 64). The “*” indicates significant correlation with adjusted P

Figure S8. Evaluation of enterotying protocols for Dutch children. (a-c) Evaluation of optimal cluster number by using the DMM protocol (a), the PAM-JSD protocol (b) and the PAM-BC protocol (c). The optimal number of clusters was calculated using Laplace approximation for the DMM protocol (a) and the Calinskiâ Harabasz index for the PAM-based protocols (b-c). Cluster stability using the DMM (d), the PAM-JSD (e) and the PAM-BC protocols (f). The X axis indicates resampling number and the Y axis indicates the consistency of resampling relative to the original result based on 281 samples. (PDF 221 kb)

Figure S5. Correlations between continuous phenotypic parameters and species profile in the entire cohort. (a) Spearman’s rank correlations between early events, pre-school lifestyle and species profile (n = 281). (b) Spearman’s rank correlations between blood parameters and species profiles (n = 281). P values were adjusted for each parameter. The “*” indicates significant correlation with adjusted P

Figure S7. Correlations between Streptococcus species profile and selected phenotypic parameters in enterotypes. Heatmap showing the Spearman’s rank correlations between Streptococcus species and selected phenotypic parameters including free fatty acids levels and the intake of total carbohydrate, total fat, dietary fiber, and plant-based protein. P values were adjusted for each parameter. The “*” indicates significant correlation with adjusted P 0.05. FFA, free fatty acids. (PDF 197 kb)

Figure S3. Comparison of gut microbial compositional and functional structure between enterotypes. (a-c) Comparison of gene count, gene-based Shannon diversity and reads mapping ratio to the taxonomic annotated genes between enterotypes. (d-f) Comparison of gene count, KO-based Shannon diversity and reads mapping ratio to the KO annotated genes between enterotypes. Dunn’s post hoc test, *, P

The gut microbiota not only influences host metabolism but can also affect brain function and behaviour through the microbiota-gut-brain axis. To explore the potential role of the intestinal microbiota in anorexia nervosa (AN), we comprehensively investigated the faecal microbiota and short-chain fatty acids in these patients before (n = 55) and after weight gain (n = 44) in comparison to normal-weight participants (NW, n = 55) along with dietary intake and gastrointestinal complaints. We show profound microbial perturbations in AN patients as compared to NW participants, with higher levels of mucin-degraders and members of Clostridium clusters I, XI and XVIII and reduced levels of the butyrate-producing Roseburia spp. Branched-chain fatty acid concentrations, being markers for protein fermentation, were elevated. Distinct perturbations in microbial community compositions were observed for individual restrictive and binge/purging AN-subtypes. Upon weight gain, microbial richness increased, however perturbations in intestinal microbiota and short chain fatty acid profiles in addition to several gastrointestinal symptoms did not recover. These insights provide new leads to modulate the intestinal microbiota in order to improve the outcomes of the standard therapy.

Aim: The aim was to study acquisition and persistence of carbapenemase-producing Enterobacteriaceae (CPE) among travelers. Materials & methods: Stools from 2001 travelers and 215 nontraveling household members, collected before and immediately post-travel as well as 1, 3, 6 and 12 months upon return, were screened for CPE. Results: Five travelers, all visiting Asia outside the Indian subcontinent, acquired CPE. One traveler persistently carried the same OXA-244 CPE up to 6 months post-travel. Three months after travel, her co-traveling spouse also became positive for this OXA-244 CPE strain, suggesting clonal transmission within this household. Conclusion: Acquisition of CPE is not restricted to travelers to the Indian subcontinent and/or to travelers seeking healthcare during travel and can persist up to at least 6 months post-travel.

Introduction: A lack of prospective and longitudinal data on pre- and post-travel carriage of Blastocystis spp. complicates interpretation of a positive WA post-travel. Therefore we studied dynamics of Blastocystis carriage in a cohort of Dutch travellers.Methods: From the prospective, multicentre COMBAT study among 2001 Dutch travellers, a subset of 491 travellers was selected based on travel destination to 7 subregions (70 or 71 travellers each). Faecal samples taken directly before and after travel were screened for Blastocystis with qPCR, followed, when positive, by sequence analysis to determine subtypes.Results: After exclusion of 12 samples with missing samples or inhibited qPCR-reactions, stool samples of 479 travellers were analysed. Before travel, 174 of them (36.3%) carried Blastocystis and in most of these, the same subtype was persistently carried. However, in 48/174 of those travellers (27.6%; CI95 20.8-36.6%) no Blastocystis or a different subtype was detected in the post-travel sample, indicating loss of Blastocystis during travel. Only 26 (5.4%; CI95 3.7%-8.0%) of all travellers acquired Blastocystis, including two individuals that were already positive for Blastocystis before travel but acquired a different subtype during travel.Discussion: This study shows that Blastocystis carriage in travellers is highly dynamic. The observed acquisition and loss of Blastocystis could either be travel-related or reflect the natural course of Blastocystis carriage. We demonstrate that the majority of Blastocystis detected in post-travel samples were already carried before travel.