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Abstract In Bangladesh, Vibrio cholerae lineages are undergoing genomic evolution, with increased virulence and spreading ability. However, our understanding of the genomic determinants influencing lineage transmission and disease severity remains incomplete. Here, we developed a computational framework using machine-learning, genome scale metabolic modelling (GSSM) and 3D structural analysis, to identify V. cholerae genomic traits linked to lineage transmission and disease severity. We analysed in-patients isolates from six Bangladeshi regions (2015-2021), and uncovered accessory genes and core SNPs unique to the most recent dominant lineage, with virulence, motility and bacteriophage resistance functions. We also found a strong correlation between V. cholerae genomic traits and disease severity, with some traits overlapping those driving lineage transmission. GSMM and 3D structure analysis unveiled a complex interplay between transcription regulation, protein interaction and stability, and metabolic networks, associated to lifestyle adaptation, intestinal colonization, acid tolerance and symptom severity. Our findings support advancing therapeutics and targeted interventions to mitigate cholera spread.

Objectives: The severity of the diarrhea disease is exacerbated by co-infections that involve Enterotoxigenic Escherichia coli (ETEC) and other enteric pathogens, which complicate the diagnosis and treatment. This study explores the prevalence, clinical manifestations, and risk factors of ETEC and its co-infections in Dhaka, Bangladesh. Methods: The study used data from the Diarrheal Disease Surveillance System at Dhaka Hospital, involving 16,276 patients from 2017 to 2022. We identified 1404 ETEC-positive patients from the 16,276 data points to investigate the association between ETEC infections, co-infections, and clinical outcomes. Results: ETEC was identified in 1404 (8.6%) of cases, representing the most common infection among adults (56.6%). ETEC co-infection occurred combined with V. cholerae (23%), Aeromonas (14.6%), rotavirus (11.8%), Campylobacter (6.5%), and Shigella spp. (1.7%), respectively. Adults were more likely to develop co-infections with ETEC and V. cholerae, while children under five were more likely to develop ETEC co-infections with rotavirus. Co-infections with V. cholerae, rotavirus, and Salmonella spp. increased the likelihood of fever, while ETEC co-infections with V. cholerae increased risks of vomiting, dehydration, and intravenous fluids. Conclusions: ETEC and co-infections exacerbate illness severity and overburden healthcare systems. Policymakers should prioritize resilient healthcare strategies for ETEC and co-infections.

BackgroundAge plays a significant role in susceptibility to enterotoxigenic Escherichia coli (ETEC) infections, yet the distribution of ETEC virulence factors across age groups remains understudied. This study investigated the differential pathogenic profiles ETEC across various age groups, emphasizing the importance of selecting potential ETEC antigens tailored to infection patterns in infants and adults in Bangladesh.MethodsThis study utilized the icddr,b’s 2% systematic hospital surveillance data of diarrheal patients (n = 14,515) from 2017 to 2022 to examine the age-specific pathogenesis and clinical manifestations of ETEC infections.ResultsIn total ETEC was identified in 1,371 (9.4%) of surveillance samples. ETEC-associated diarrhea was higher in children aged 0–2 years and decreased significantly in the 3–17 years age group. Among all ETEC cases, 56% were adults (p = 0.0079) with severe dehydration. Distinct age-specific distribution of ETEC toxin types and colonization factors (CFs) were observed: heat labile toxin (LT)-associated ETEC infections decreased with age (p

ABSTRACT The objective of this study was to assess the prevalence, antimicrobial resistance patterns, and risk factors linked to Shigella infections through a nationwide hospital-based diarrheal disease surveillance in Bangladesh. From May 2014 to May 2022, from a systematic sentinel surveillance of Shigella infections in over 10 hospitals across Bangladesh, stool specimens were collected from patients with acute watery diarrhea and tested for Shigella species by microbiological culture. The susceptibility to antibiotics was tested using the disk diffusion method. Structured questionnaires were used to collect participants’ socioeconomic status and clinical, sanitation, and food history. Out of 24,357 stool specimens, 1.8% were positive for Shigella species, with a higher prevalence among males (58%). Children in the 6–17 age group were found to be at the highest risk of Shigella infections. The most prevalent serotype was Shigella flexneri (79.5%), followed by Shigella sonnei. S. flexneri serotype 2a was the most common (63.3%) among all Shigella serotypes. Antibiotic susceptibility testing showed over 99% of isolates resistant or with intermediate susceptibility to one of the seven antibiotics tested. About 96% of S. flexneri and all S. sonnei isolates demonstrated resistance to at least one quinolone class of antibiotics, particularly ciprofloxacin or nalidixic acid. S. sonnei showed higher antibiotic resistance and multidrug resistance compared to S. flexneri. The high level of resistance to ciprofloxacin highlights the need for more prudent use of this antibiotic and improved hygiene and sanitation. The study emphasized the importance of regular monitoring of drug resistance to effectively manage Shigella infections. These findings may provide the epidemiological evidence for conducting future appropriate Shigella vaccine clinical trials in Bangladesh.IMPORTANCEThis nationwide study in Bangladesh assessed Shigella infections from 2014 to 2022 from clinical samples. S. flexneri was predominant, with concerning antibiotic resistance, notably to ciprofloxacin and nalidixic acid in over 96% of isolates. This emphasizes the urgency of prudent antibiotic use and improved hygiene. The findings provide crucial antimicrobial resistance patterns of Shigella species, highlighting the need for ongoing resistance monitoring and potentially informing future vaccine trials.

Diarrheal diseases, especially cholera, can be a serious threat to Rohingya refugees in Cox's Bazar due to overcrowding and inadequate hygiene infrastructure. Assessing the risk, cholera surveillance network was established with the aim to identify the outbreak of diarrhea and cholera and help to take appropriate preventive measures including a vaccination campaign. The surveillance network has been ongoing for 6 years (2017–2023) in 17 health facilities. Diarrhea patients from Rohingya Myanmar nationals matched with case definition were enrolled and stool samples were tested by Rapid diagnostic test (RDT) for early cholera detection Multiple Logistic regression models were fitted to examine the associations of risk factors among cholera cases. A total of 17,252 stool samples were collected through surveillance. Among the tested samples, 588 (3.5 %) were detected positive by RDT, and 239 (1.4 %) Vibrio cholerae were isolated by microbiological culture. Between 2021 and 2023, the number of culture-confirmed cases exceeded that in the period from 2017 to 2020. In addition to V. cholerae; high positivity was identified for ETEC (11.8 %) followed by Salmonella (3.9 %) and Shigella (2.7 %). Most of the cholera cases were presented with vomiting, dehydration and loose watery and rice watery nature of stool (p value =

Introducción. El “moho verde” causado por Penicillium digitatum es uno de los problemas fitosanitarios que limita la producción en los cítricos. Para su control se utilizan fungicidas sintéticos, que han perdido eficacia, debido a la resistencia generada por el hongo. Una alternativa, es el uso de hongos endófitos con acción biocontroladora. Objetivo. Evaluar el efecto de metabolitos secundarios (MS) de hongos endófitos (HE) en la inhibición micelial de Penicillium digitatum y la eficacia de control de la enfermedad, en frutos de mandarina (Citrus reticulata) después de la cosecha. Materiales y métodos. El estudio se realizó en 2022, bajo un diseño experimental completo al azar, en el laboratorio de Sanidad Vegetal, Universidad Nacional de Cañete (UNDC), Lima, Perú. Para la prueba de antibiosis se extrajeron los metabolitos secundarios de diez cepas de HE de los géneros Trichoderma sp., Fusarium sp. y Aspergillus sp., en donde se evaluó la inhibición micelial del patógeno (%). Se asperjaron MS a una concentración del 15 % (v/v) sobre frutos de mandarina que fueron desinfectados e inoculados con el patógeno por aspersión (1x105 UFC/cc). Adicionalmente, se utilizaron un biocontrol comercial, un aceite vegetal y un fungicida. Después de 6 y 9 días, se evaluaron la incidencia (%) y el índice de severidad (0,1,2,3) de la enfermedad. Resultados. Los metabolitos secundarios de las cepas HEA-111 (Aspergillus), HEA-109 (Fusarium) y HSA-1 (Trichoderma) presentaron la mayor inhibición micelial con 95,05 %, 87,51 % y 47,47 %, respectivamente. En los frutos de mandarina tratados con los MS de las cepas HEA-109 y HSA-1, hubo un control del 80 % y 55 %, respectivamente. Conclusiones. Los MS de HE tuvieron efecto en la inhibición micelial de P. digitatum y redujeron el avance de la enfermedad en frutos de mandarina después de la cosecha.

Currently approved repetitive transcranial magnetic stimulation (rTMS) protocols for the treatment of major depressive disorder (MDD) involve once-daily (weekday) stimulation sessions, with 10 Hz or intermittent theta burst stimulation (iTBS) frequencies, over 4–6 weeks. Recently, accelerated treatment protocols (multiple daily stimulation sessions for 1–2 weeks) have been increasingly studied to optimize rTMS treatments. Accelerated protocols might confer unique advantages for adolescents and young adults but there are many knowledge gaps related to dosing in this age group. Off-label, clinical practice frequently outpaces solid evidence as rigorous clinical trials require substantial time and resources. Murine models present an opportunity for high throughput dose finding studies to focus subsequent clinical trials in humans. This project investigated the brain and behavioural effects of an accelerated low-intensity rTMS (LI-rTMS) protocol in a young adult rodent model of chronic restraint stress (CRS). Depression and anxiety-related behaviours were induced in young adult male Sprague Dawley rats using the CRS model, followed by the 3-times-daily delivery of 10 Hz LI-rTMS, for two weeks. Behaviour was assessed using the Elevated Plus Maze and Forced Swim Test, and functional, chemical, and structural brain changes measured using magnetic resonance imaging techniques. CRS induced an agitated depression-like phenotype but therapeutic effects from the accelerated protocol were not detected. Our findings suggest that the age of rodents may impact response to CRS and LI-rTMS. Future studies should also examine higher intensities of rTMS and accelerated theta burst protocols.

Carbon nanomaterials, including carbon dots (CDs), form a growing family of engineered nanoparticles (NPs) with widespread applications. As the rapid expansion of nanotechnologies raises safety concerns, interaction of NPs with the immune system is receiving a lot of attention. Recent studies have reported that engineered NPs may induce macrophage death by pyroptosis. Therefore, this study investigated whether cationic CDs induce pyroptosis in human macrophages and assessed the role of inflammasome and lysosome in this process. Cationic CDs were synthetized by microwave-assisted pyrolysis of citric acid and high molecular weight branched polyethyleneimine. The NPs evoked a dose-dependent viability loss in THP-1-derived macrophages. A cell leakage, an increase in IL-1β secretion and an activation of caspase-1 were also observed in response to the NPs. Inhibition of caspase-1 decreased CD-induced cell leakage and IL-1β secretion, while restoring cell viability. Besides, CDs triggered swelling and loss of integrity of lysosome, and inhibition of the lysosomal enzyme cathepsin B decreased CD-induced IL-1β secretion. Thus, our data provide evidence that cationic CDs induce inflammasome-dependent pyroptosis in macrophages via lysosomal dysfunction.

Gut microbiome composition is associated with mood-relating behaviours, including those reflecting depression-like phenotypes. Repetitive transcranial magnetic stimulation (rTMS), a non-invasive neuromodulation technique, is an effective treatment for depression, but its effects on the gut microbiome remain largely unknown. This study assessed microbial changes from rat faecal samples longitudinally following chronic restraint stress (CRS) and 10 Hz low-intensity rTMS treatment. CRS increased abundance within the Proteobacteria (Deltaproteobacteria, Desulfovibrionales) and Firmicutes (Anaerostipes, Frinsingococcus), with decreases in Firmicutes family (Acidaminococcaceae) and genera (Roseburia, Phascolarctobacterium and Fusicatenibacter) persisting for up to 4 weeks post CRS. The decrease in Firmicutes was not observed in the handling control and LI-rTMS groups, suggesting that handling alone may have sustained changes in gut microbiome associated with CRS. Nonetheless, LI-rTMS was specifically associated with an increase in Roseburia genus that developed 2 weeks after treatment, and the abundance of both Roseburia and Fusicatenibacter genera was significantly correlated with rTMS behavioural and MRI outcomes. In addition, LI-rTMS treated rats had a reduction in apoptosis pathways and several indicators of reduced inflammatory processes. These findings provide evidence that the brain can influence the gut microbiome in a “top-down” manner, presumably via stimulation of descending pathways, and/or indirectly via behavioural modification.

Major cities in Indonesia generally have landmarks or special features that symbolize the uniqueness and beauty of the city in question. One of the most important landmarks of the colonial city is the societeit building as an entertainment center. This study examines the function and role of Societeit de Harmonie in Batavia in the 19th century in the midst of the heterogeneous and multiethnic life of the Batavian city of Batavia. Through the study of documents and archives, the results of this study show that Societeit de Harmonie is the greatest building compared to other societeit buildings in Batavia in the 19th century. Therefore, this building is known by all citizens of the city although who can access and utilize this building is only limited. Only the European elite can enter and use the Societeit de Harmonie building. The presence of the place of entertainment also represents the entry of new values (West) that color and becomes an important part of the cultural development of the next city Batavia. These Western values are coupled with local values practiced by indigenous peoples living in the corners of the city of Batavia. The lifestyles of Europeans who lived in Batavia during the nineteenth century can be identified from their activities when gathered in the Societeit de Harmonie. Kota-kota besar di Indonesia pada umumnya memiliki landmark atau ciri khusus yang menjadi lambang keunikan dan keindahan kota yang bersangkutan. Salah satu landmark kota colonial yang terpenting adalah gedung societeit sebagai pusat hiburan. Penelitian ini mengkaji fungsi dan peran Societeit de Harmonie di Batavia pada abad ke-19 di tengah-tengah kehidupan masyarakat kota Batavia yang heterogen dan multietnis. Melalui studi dokumen dan arsip-arsip sezaman, hasil penelitian ini menunjukkan bahwa Societeit de Harmoni merupakan bangunan termegah dibandingkan bangunan societeit lainnya di Batavia pada abad ke-19. Oleh karena itu, bangunan ini dikenal oleh seluruh warga kota meskipun yang dapat mengakses dan memanfaatkan bangunan ini hanya kalangan terbatas. Hanya golongan elit Eropa saja yang dapat masuk dan menggunakan gedung Societeit de Harmonie. Kehadiran tempat hiburan itu juga merepresentasikan masuknya nilai-nilai baru (Barat) yang mewarnai dan menjadi bagian penting dari perkembagan budaya kota Batavia selanjutnya. Nilai-nilai Barat ini bersanding dengan nilai-nilai local yang dipraktikkan masyarakat pribumi yang tinggal di sudut- sudut kota Batavia. Gaya hidup orang-orang Eropa yang tinggal di Batavia selama abad ke-19 dapat dikenali dari aktivitas mereka ketika berkumpul di Societeit de Harmonie.

Background: Immune thrombotic thrombocytopenic purpura (iTTP) is a rare, life-threatening disorder managed with plasma exchange (PLEX) and steroids. Addition of rituximab (RTX) to initial disease treatment has been shown to lower future relapse rates. Information as to whether upfront cyclophosphamide (CTX) treatment is helpful in reducing relapse is not known.Methods: In a retrospective cohort study, we identified all patients at our institution diagnosed with iTTP between 2010 and 2019. We analyzed outcomes of cumulative incidence of relapse (CIR) and duration of remission.Results: Thirty Nine patients were studied. Group A (n = 10) included patients who received upfront PLEX and steroids alone, and Group B (n = 28) included those who received either upfront RTX (n = 23) or CTX (n = 5) in addition to PLEX and steroids. The 2-year CIR was 50% in Group A and 27.7% in Group B, with a median duration of remission of 43.6 months vs. 108.3 months, respectively (p = 0.04). Group A was associated with a HR=8.7 (95% CI: 1.27, 59.45), p = 0.027 for duration of remission. There was no significant difference between CTX and RTX in both outcomes of CIR and duration of remission. We observed a potential impact on remission duration based on the presenting absolute neutrophil count (HR = 0.74, 95% CI: 0.58, 0.96) and serum creatinine (HR = 1.42, 95% CI: 1.03, 1.94).Conclusion: There was no significant difference in iTTP relapse outcomes between upfront RTX and CTX. Absolute neutrophil count and serum creatinine may have a role in predicting relapse. Larger, prospective studies are needed to evaluate these findings.

This study analyzes the main aspects of the stabilization of the anticipated guardianship required in an antecedent character brought by the Civil Procedure Code of 2015, highlighting the possibility to argue the material law as a matter of defense after the passage in albis of the two years term. Initially, its origins will be analyzed so that the technique that influenced this institute will be identified, and the study of its presuppositions and controversial points in the doctrine will be studied. After confirming its requirements and the legal nature of the decision, the study faces the doctrinal controversies about the modification of the anticipatory decision after the lapse of two years.

The present paper aims to present a proposal to incorporate sustainability and social responsibility in an apparel production company. The article describes the importance of the topic, as well as literature reviews about sustainability and the fashion chain in Brazil, and proposes a cohesive and effective model for the reuse of fabric leftovers in a manufacturing industry by using Bomfim’s (1975) product design methodology.

A 15-year-old boy 4th issue of a non-consanguineous parents presented with a history of multiple plaques and patches on the scalp, trunk, upper and lower extremities since birth. Initially, the plaques and patches were smaller in sizes, dark brown in color, painless and non-pruritic. Since 4th year of his age, these plaques start increasing in size and color turned into black gradually. The lesion on the scalp became a giant cerebriform plaque that involved left parietal region. Lesions on the trunk, upper and lower extremities became polysized and polyshaped hyperpigmented papules and nodules.

This study aimed at discriminating with sensitivity the toxicological effects of carbon dots (CDs) with various zeta potential (ζ) and charge density (Qek) in different cellular models of the human respiratory tract. One anionic and three cationic CDs were synthetized as follows: CD-COOH (ζ = −43.3 mV); CD-PEI600 (Qek = 4.70 µmol/mg; ζ = +31.8 mV); CD-PEHA (Qek = 3.30 µmol/mg; ζ = +29.2 mV) and CD-DMEDA (Qek = 0.01 µmol/mg; ζ = +11.1 mV). Epithelial cells (A549) and macrophages (THP-1) were seeded alone or as co-cultures with different A549:THP-1 ratios. The obtained models were characterized, and multiple biological responses evoked by CDs were assessed in the mono-cultures and the best co-culture model. With 14% macrophages, the 2:1 ratio co-culture best mimicked the in vivo conditions and responded to lipopolysaccharides. The anionic CD did not induce any effect in the mono-cultures nor in the co-culture. Among the cationic CDs, the one with the highest charge density (CD-PEI600) induced the most pronounced responses whatever the culture model. The cationic CDs of low charge density (CD-PEHA and CD-DMEDA) evoked similar responses in the mono-cultures, whereas in the co-culture, the three cationic CDs ranked according to their charge density (CD-PEI600 > CD-PEHA > CD-DMEDA), when taking into account their inflammatory effect. Thus, the co-culture system developed in this study appears to be a sensitive model for finely discriminating the toxicological profile of cationic nanoparticles differing by the density of their surface charges.

[This corrects the article DOI: 10.1371/journal.pone.0209357.].

BACKGROUND:Cholera remains a substantial health burden in Asia and Africa particularly in resource poor settings. The standard procedures to identify the etiological organism V. cholerae are isolation from microbiological culture from stool as well as Polymerase Chain Reaction (PCR). Both the processes are highly lab oriented, labor extensive, time consuming, and expensive. In an effort to control for outbreaks and epidemics; an effective, convenient, quick and relatively less expensive detection method is imperative, without compromising the sensitivity and specificity that exists at present. The objective of this component of the study was to evaluate the effectiveness of a locally produced rapid diagnostic test (RDT) for cholera diagnosis. METHODS:In Bangladesh, nationwide cholera surveillance is ongoing in 22 hospitals covering all 8 divisions of the country since June, 2016. In the surveillance, stool samples have been collected from patients presenting to hospitals with acute watery diarrhea. Crystal VCTM (Span diagnostics, India) and Cholkit (locally produced RDT) have been used to detect V. cholerae from stool samples. Samples have also been sent to the main laboratory at icddr,b where the culture based isolation is routinely performed. All the tests were carried out for both direct and enriched stool samples. RDT sensitivity and specificity were calculated using stool culture as the gold standard. RESULTS:A total of 7720 samples were tested. Among these, 5865 samples were solely tested with Crystal VC and 1355 samples with Cholkit whereas 381 samples were tested with both the RDTs. In comparison with culture, direct testing with Crystal VC showed a sensitivity of 72% (95% CI: 50.6% to 87.9%) and specificity of 86.8% (95% CI: 82.8% to 90.1%). After enrichment the sensitivity and specificity was 68% (95% CI: 46.5% to 85.1%) and 97.5% (95% CI: 95.3% to 98.8%) respectively. The direct Cholkit test showed sensitivity of 76% (95% CI: 54.9% to 90.6%) and specificity of 90.2% (95% CI: 86.6% to 93.1%). CONCLUSION:This evaluation has demonstrated that the sensitivity and specificity of Cholkit is similar to the commercially available test, Crystal VC when used in field settings for detecting V. cholerae from stool specimens. The findings from this study suggest that the Cholkit could be a possible alternative for cholera endemic regions where V. cholerae O1 is the major causative organism causing cholera.

ABSTRACT The bacterial pathogens enterotoxigenic Escherichia coli (ETEC) and Vibrio cholerae are major causes of diarrhea. ETEC causes diarrhea by production of the heat-labile toxin (LT) and heat-stable toxins (STh and STp), while V. cholerae produces cholera toxin (CT). In this study, we determined the occurrence and bacterial doses of the two pathogens and their respective toxin expression levels directly in liquid diarrheal stools of patients in Dhaka, Bangladesh. By quantitative culture and real-time quantitative PCR (qPCR) detection of the toxin genes, the two pathogens were found to coexist in several of the patients, at concentrations between 102 and 108 bacterial gene copies per ml. Even in culture-negative samples, gene copy numbers of 102 to 104 of either ETEC or V. cholerae toxin genes were detected by qPCR. RNA was extracted directly from stool, and gene expression levels, quantified by reverse transcriptase qPCR (RT-qPCR), of the genes encoding CT, LT, STh, and STp showed expression of toxin genes. Toxin enzyme-linked immunosorbent assay (ELISA) confirmed active toxin secretion directly in the liquid diarrhea. Analysis of ETEC isolates by multiplex PCR, dot blot analysis, and genome sequencing suggested that there are genetic ETEC profiles that are more commonly found as dominating single pathogens and others that are coinfectants with lower bacterial loads. The ETEC genomes, including assembled genomes of dominating ETEC isolates expressing LT/STh/CS5/CS6 and LT/CS7, are provided. In addition, this study highlights an emerging important ETEC strain expressing LT/STp and the novel colonization factor CS27b. These findings have implications for investigations of pathogenesis as well as for vaccine development. IMPORTANCE The cause of diarrheal disease is usually determined by screening for several microorganisms by various methods, and sole detection is used to assign the agent as the cause of disease. However, it has become increasingly clear that many infections are caused by coinfections with several pathogens and that the dose of the infecting pathogen is important. We quantified the absolute numbers of enterotoxigenic E. coli (ETEC) and Vibrio cholerae directly in diarrheal fluid. We noted several events where both pathogens were found but also a large dose dependency. In three samples, we found ETEC as the only pathogen sought for. These isolates belonged to globally distributed ETEC clones and were the dominating species in stool with active toxin expression. This suggests that certain superior virulent ETEC lineages are able to outcompete the gut microbiota and be the sole cause of disease and hence need to be specifically monitored.

Diarrhea due to infection of enterotoxigenic Escherichia coli (ETEC) is of great concern in several low and middle-income countries. ETEC infection is considered to be the most common cause of diarrhea in Bangladesh and is mainly spread through contaminated water and food. ETEC pathogenesis is mediated by the expression of enterotoxins and colonization factors (CFs) that target the intestinal mucosa. ETEC can survive for extended time periods in water, where they are likely to be attacked by bacteriophages. Antibiotic resistance is common amongst enteric pathogens and therefore is the use of bacteriophages (phage) as a therapeutic tool an interesting approach. This study was designed to identify novel phages that specifically target ETEC virulence factors. In total, 48 phages and 195 ETEC isolates were collected from water sources and stool samples. Amongst the identified ETEC specific phages, an enterobacteria phage T7, designated as IMM-002, showed a significant specificity towards colonization factor CS3-expressing ETEC isolates. Antibody-blocking and phage-neutralization assays revealed that CS3 is used as a host receptor for the IMM-002 phage. The bacterial CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-associated) defence mechanism can invoke immunity against phages. Genomic analyses coupled with plaque assay experiments indicate that the ETEC CRISPR-Cas system is involved in the resistance against the CS3-specific phage (IMM-002) and the previously identified CS7-specific phage (IMM-001). As environmental water serves as a reservoir for ETEC, it is important to search for new antimicrobial agents such as phages in environmental water as well as the human gut. A better understanding of how the interplay between ETEC-specific phages and ETEC isolates affects the ETEC diversity, both in environmental ecosystems and within the host, is important for the development of new treatments for ETEC infections.

Enterotoxigenic Escherichia coli (ETEC), defined by their elaboration of heat-labile (LT) and/or heat-stable (ST) enterotoxins, are a common cause of diarrheal illness in developing countries. Efficient delivery of these toxins requires ETEC to engage target host enterocytes. This engagement is accomplished using a variety of pathovar-specific and conserved E. coli adhesin molecules as well as plasmid encoded colonization factors. Some of these adhesins undergo significant transcriptional modulation as ETEC encounter intestinal epithelia, perhaps suggesting that they cooperatively facilitate interaction with the host. Among genes significantly upregulated on cell contact are those encoding type 1 pili. We therefore investigated the role played by these pili in facilitating ETEC adhesion, and toxin delivery to model intestinal epithelia. We demonstrate that type 1 pili, encoded in the E. coli core genome, play an essential role in ETEC virulence, acting in concert with plasmid-encoded pathovar specific colonization factor (CF) fimbriae to promote optimal bacterial adhesion to cultured intestinal epithelium (CIE) and to epithelial monolayers differentiated from human small intestinal stem cells. Type 1 pili are tipped with the FimH adhesin which recognizes mannose with stereochemical specificity. Thus, enhanced production of highly mannosylated proteins on intestinal epithelia promoted FimH-mediated ETEC adhesion, while conversely, interruption of FimH lectin-epithelial interactions with soluble mannose, anti-FimH antibodies or mutagenesis of fimH effectively blocked ETEC adhesion. Moreover, fimH mutants were significantly impaired in delivery of both heat-stable and heat-labile toxins to the target epithelial cells in vitro, and these mutants were substantially less virulent in rabbit ileal loop assays, a classical model of ETEC pathogenesis. Collectively, our data suggest that these highly conserved pili play an essential role in virulence of these diverse pathogens.

BACKGROUND:To take advantage of emerging opportunities to reduce morbidity and mortality from diarrheal disease, we need to better understand the determinants of life-threatening severe dehydration (SD) in resource-poor settings. METHODOLOGY/FINDINGS:We analyzed records of patients admitted with acute diarrheal disease over twenty-two years at the International Centre for Diarrhoeal Disease Research, Bangladesh (1993-2014). Patients presenting with and without SD were compared by multivariable logistic regression models, which included socio-demographic factors and pathogens isolated. Generalized additive models evaluated non-linearities between age or household income and SD. Among 55,956 admitted patients, 13,457 (24%) presented with SD. Vibrio cholerae was the most common pathogen isolated (12,405 patients; 22%), and had the strongest association with SD (AOR 4.77; 95% CI: 4.41-5.51); detection of multiple pathogens did not exacerbate SD risk. The highest proportion of severely dehydrated patients presented in a narrow window only 4-12 hours after symptom onset. Risk of presenting with SD increased sharply from zero to ten years of age and remained high throughout adolescence and adulthood. Adult women had a 38% increased odds (AOR 1.38; 95% CI: 1.30-1.46) of SD compared to adult men. The probability of SD increased sharply at low incomes. These findings were consistent across pathogens. CONCLUSIONS/SIGNIFICANCE:There remain underappreciated populations vulnerable to life-threatening diarrheal disease that include adult women and the very poor. In addition to efforts that address diarrheal disease in young children, there is a need to develop interventions for these other high-risk populations that are accessible within 4 hours of symptom onset.

BACKGROUND:Cholera is endemic in Bangladesh, with outbreaks reported annually. Currently, the majority of epidemic cholera reported globally is El Tor biotype Vibrio cholerae isolates of the serogroup O1. However, in Bangladesh, outbreaks attributed to V. cholerae serogroup O139 isolates, which fall within the same phylogenetic lineage as the O1 serogroup isolates, were seen between 1992 and 1993 and in 2002 to 2005. Since then, V. cholerae serogroup O139 has only been sporadically isolated in Bangladesh and is now rarely isolated elsewhere. METHODS:Here, we present case histories of four cholera patients infected with V. cholerae serogroup O139 in 2013 and 2014 in Bangladesh. We comprehensively typed these isolates using conventional approaches, as well as by whole genome sequencing. Phenotypic typing and PCR confirmed all four isolates belonging to the O139 serogroup. FINDINGS:Whole genome sequencing revealed that three of the isolates were phylogenetically closely related to previously sequenced El Tor biotype, pandemic 7, toxigenic V. cholerae O139 isolates originating from Bangladesh and elsewhere. The fourth isolate was a non-toxigenic V. cholerae that, by conventional approaches, typed as O139 serogroup but was genetically divergent from previously sequenced pandemic 7 V. cholerae lineages belonging to the O139 or O1 serogroups. CONCLUSION:These results suggest that previously observed lineages of V. cholerae O139 persist in Bangladesh and can cause clinical disease and that a novel disease-causing non-toxigenic O139 isolate also occurs.

Flooding in Dhaka in July 2004 caused epidemics of diarrhea. Enterotoxigenic Escherichia coli (ETEC) was almost as prevalent as Vibrio cholerae O1 in diarrheal stools. ETEC that produced heat-stable enterotoxin alone was most prevalent, and 78% of strains had colonization factors. Like V. cholerae O1, ETEC can cause epidemic diarrhea.

Enterotoxigenic Escherichia coli (ETEC) is one of the most common causes of bacterial diarrhea. Over the last decade, from 1996 to 2012, changes in the virulence antigen properties of ETEC such as heat labile (LT) and heat stable (ST) toxins, colonization factors (CFs), and 'O'-serogroups have been observed. The aim of this prospective study was to compare changes in antigenic profiles of ETEC strains isolated from a 2% surveillance system at the icddr,b hospital in Dhaka, Bangladesh between 2007-2012 and an earlier time period of 1996-1998 conducted at the same surveillance site.In the surveillance system every 50th patient attending the hospital was screened for major enteric pathogens including ETEC, Vibrio cholerae, Shigella spp. and Salmonella spp. from January 2007 to December 2012.Of the 15,152 diarrheal specimens tested between 2007-2012, the overall rate of ETEC isolation was 11%; of these, 43% were LT/ST, 27% LT and 30% ST positive. Isolation rate of ST-ETEC (p

BackgroundMultiple infections with diverse enterotoxigenic E. coli (ETEC) strains lead to broad spectrum protection against ETEC diarrhea. However, the precise mechanism of protection against ETEC infection is still unknown. Therefore, memory B cell responses and affinity maturation of antibodies to the specific ETEC antigens might be important to understand the mechanism of protection.MethodologyIn this study, we investigated the heat labile toxin B subunit (LTB) and colonization factor antigens (CFA/I and CS6) specific IgA and IgG memory B cell responses in Bangladeshi adults (n = 52) who were infected with ETEC. We also investigated the avidity of IgA and IgG antibodies that developed after infection to these antigens.Principal findingsPatients infected with ETEC expressing LT or LT+heat stable toxin (ST) and CFA/I group or CS6 colonization factors developed LTB, CFA/I or CS6 specific memory B cell responses at day 30 after infection. Similarly, these patients developed high avidity IgA and IgG antibodies to LTB, CFA/I or CS6 at day 7 that remained significantly elevated at day 30 when compared to the avidity of these specific antibodies at the acute stage of infection (day 2). The memory B cell responses, antibody avidity and other immune responses to CFA/I not only developed in patients infected with ETEC expressing CFA/I but also in those infected with ETEC expressing CFA/I cross-reacting epitopes. We also detected a significant positive correlation of LTB, CFA/I and CS6 specific memory B cell responses with the corresponding increase in antibody avidity.ConclusionThis study demonstrates that natural infection with ETEC induces memory B cells and high avidity antibodies to LTB and colonization factor CFA/I and CS6 antigens that could mediate anamnestic responses on re-exposure to ETEC and may help in understanding the requirements to design an effective vaccination strategies.