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Background: Globally, gastroenteritis (GE) significantly impacts children's health and contributes to societal, economic, and health burdens. Previous studies reporting risk factors of GE in children in high-income settings mainly rely on outbreak investigations, which inherently capture only a fractional representation of the overall spectrum of GE occurrences. In addition, there is paucity of comprehensive information pertaining to modifiable risk factors of GE. This scoping review aims to synthesize existing evidence concerning modifiable and behavioural risk factors associated with GE among children in high-income countries. Methods: PubMed, Embase, CINAHL, and Scopus were the databases from which articles were retrieved. A descriptive synthesis of the evidence was performed, following the Arksey and O'Malley scoping studies framework and enhanced by the Preferred Reporting Items for Systematic Reviews and Meta-analysis extension for Scoping Reviews checklist (PRISMA-ScR). Results: The systematic search identified 13,395 journal articles, which were subsequently screened, and duplicates removed, resulting in 19 articles for inclusion in the review. The majority of these studies (63.2%) employed a case-control design and were predominantly conducted in community settings (68.4%). Factors such as parental literacy, contact with individuals exhibiting gastrointestinal symptoms, and nappy-wearing were identified as significantly associated with childhood GE within domestic environments. Childcare-related variables, including enrolment size, mixing of personnel between child groups, the presence of central cleaning stations, and the implementation of hygiene and disease prevention policies, showed significant association with GE. In addition, the presence of sand pits, paddling pools, and animals in childcare centers correlated with increased incidences of GE among attending children. Conclusions: The scoping review reveals a complex and varied research landscape on factors influencing gastroenteritis (GE) for children in high-income countries. The findings suggest that while some variables are closely linked to specific pathogens, others may not be, highlighting variability across GE aetiology. The significant association between various household level and childcare-related factors and childhood GE points to a valuable direction for future research and public health intervention. [ABSTRACT FROM AUTHOR]

Abstract Background Globally, gastroenteritis (GE) significantly impacts children’s health and contributes to societal, economic, and health burdens. Previous studies reporting risk factors of GE in children in high-income settings mainly rely on outbreak investigations, which inherently capture only a fractional representation of the overall spectrum of GE occurrences. In addition, there is paucity of comprehensive information pertaining to modifiable risk factors of GE. This scoping review aims to synthesize existing evidence concerning modifiable and behavioural risk factors associated with GE among children in high-income countries. Methods PubMed, Embase, CINAHL, and Scopus were the databases from which articles were retrieved. A descriptive synthesis of the evidence was performed, following the Arksey and O’Malley scoping studies framework and enhanced by the Preferred Reporting Items for Systematic Reviews and Meta-analysis extension for Scoping Reviews checklist (PRISMA-ScR). Results The systematic search identified 13,395 journal articles, which were subsequently screened, and duplicates removed, resulting in 19 articles for inclusion in the review. The majority of these studies (63.2%) employed a case-control design and were predominantly conducted in community settings (68.4%). Factors such as parental literacy, contact with individuals exhibiting gastrointestinal symptoms, and nappy-wearing were identified as significantly associated with childhood GE within domestic environments. Childcare-related variables, including enrolment size, mixing of personnel between child groups, the presence of central cleaning stations, and the implementation of hygiene and disease prevention policies, showed significant association with GE. In addition, the presence of sand pits, paddling pools, and animals in childcare centers correlated with increased incidences of GE among attending children. Conclusions The scoping review reveals a complex and varied research landscape on factors influencing gastroenteritis (GE) for children in high-income countries. The findings suggest that while some variables are closely linked to specific pathogens, others may not be, highlighting variability across GE aetiology. The significant association between various household level and childcare-related factors and childhood GE points to a valuable direction for future research and public health intervention.

Introduction: The definitive diagnosis of coronavirus disease-2019 (COVID-19) infection is made by polymerase chain reaction (PCR) tests on nasopharyngeal and oropharyngeal swab samples. However, the presence of viral RNA has also been identified in stool samples. In this study, we aimed to investigate the relationship between severe acute respiratory syndrome-coronavirus-2 positivity in stool and the outcomes of COVID-19 disease. Methods: Fifty-four patients who were hospitalized between April-June 2020 and had positive COVID-19 PCR tests in nasopharyngeal and oropharyngeal swab samples were included in the study. PCR was performed on the stool samples of all patients. In addition, laboratory findings, clinical data, and computed tomography (CT) results of these patients were recorded and analyzed. Results: Among the patients, 13 out of 28 (46.4%) with positive fecal PCR test results were female, whereas 11 out of 26 (46.4%) with negative fecal PCR test results were female. Furthermore, 19 out of 28 patients (67.9%) with positive fecal PCR test results recovered, whereas 23 out of 26 patients (88.5%) with negative fecal PCR test results recovered. Notably, patients with fecal PCR-positive results exhibited more severe dyspnea, higher blood pressure, abnormal CT findings, and elevated D-dimer levels. Moreover, compared with patients with negative PCR results, those with positive fecal PCR results had lower levels of procalcitonin, hemoglobin, hematocrit, and lymphocytes. Conclusion: Considering the relationship between stool PCR positivity and the prognosis of the disease and laboratory test results, routine stool PCR tests may be useful, especially in COVID-19 patients presenting with gastrointestinal symptoms.

Introduction: The definitive diagnosis of coronavirus disease-2019 (COVID-19) infection is made by polymerase chain reaction (PCR) tests on nasopharyngeal and oropharyngeal swab samples. However, the presence of viral RNA has also been identified in stool samples. In this study, we aimed to investigate the relationship between severe acute respiratory syndrome-coronavirus-2 positivity in stool and the outcomes of COVID-19 disease. Methods: Fifty-four patients who were hospitalized between April-June 2020 and had positive COVID-19 PCR tests in nasopharyngeal and oropharyngeal swab samples were included in the study. PCR was performed on the stool samples of all patients. In addition, laboratory findings, clinical data, and computed tomography (CT) results of these patients were recorded and analyzed. Results: Among the patients, 13 out of 28 (46.4%) with positive fecal PCR test results were female, whereas 11 out of 26 (46.4%) with negative fecal PCR test results were female. Furthermore, 19 out of 28 patients (67.9%) with positive fecal PCR test results recovered, whereas 23 out of 26 patients (88.5%) with negative fecal PCR test results recovered. Notably, patients with fecal PCR-positive results exhibited more severe dyspnea, higher blood pressure, abnormal CT findings, and elevated D-dimer levels. Moreover, compared with patients with negative PCR results, those with positive fecal PCR results had lower levels of procalcitonin, hemoglobin, hematocrit, and lymphocytes. Conclusion: Considering the relationship between stool PCR positivity and the prognosis of the disease and laboratory test results, routine stool PCR tests may be useful, especially in COVID-19 patients presenting with gastrointestinal symptoms. [ABSTRACT FROM AUTHOR]

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Asaye Mitiku,1 Zerihin Solomon,1 Berhanu Gidisa,1 Kasie Gebeyhu,1 Haymanot Tewabe,2 Demissew Shenkute,3 Melkayehu Kassa,4 Addisu Gize4 1Department of Medical Laboratory Science, College of Medicine and Health Sciences, Dilla University, Dilla, Ethiopia; 2Department of Medical Laboratory Science, College of Medicine and Health Sciences, Debre Markos University, Debre Markos, Ethiopia; 3Department of Medical Laboratory Science, College of Health Sciences, Debre Birhan University, Debre Berhan, Ethiopia; 4Department of Microbiology, St. Paul’s Hospital Millennium Medical College, Addis Ababa, EthiopiaCorrespondence: Asaye Mitiku, Email asayemitiku@gmail.comBackground: In people with human immunodeficiency virus infection, diarrhea is reportedly associated with significant morbidity and mortality. Therefore, the aim of this study was to determine the prevalence, antibiotic susceptibility pattern, and associated factors of enteric bacterial pathogens among HIV infected patients with diarrhea attending the antiretroviral treatment (ART) clinic of Dilla University Referral Hospital, southern Ethiopia.Methods: This institutional-based cross-sectional study was conducted on 422 study participants attending at ART clinic of Dilla University Referral Hospital from March to August 2022. Demographic and clinical data were collected by using a semi-structured questionnaire. Stool specimens were inoculated on selective media like Butzller’s medium and Xylose Lysine Deoxycholate (XLD) agar. Antimicrobial resistance pattern was assessed by using Kirby-Bauer disk diffusion techniques. Adjusted odds ratio (AOR) and 95% Confidence Interval (CI) was used to determine the presence of association.Results: A total of 422 adult patients were enrolled in this study, 51.7% were females. The mean age of the study participants was 27.4 (± 15.6 SD) years. The overall prevalence of enteric pathogens was 14.7% (95% CI=11.4– 18.2). Shigella spp was the most prevalent organism. Being a farmer (AOR=5.1; 95% CI=1.4– 19.1; p< 0.015), the habit of hand washing after toilet (AOR=1.9; 95% CI=1.02– 3.47; p< 0.04), low CD4 cell count of < 200 cells (AOR=2.22; 95% CI=1.15– 4.27; p< 0.02), and longer duration of diarrhea (AOR=2.68; 95% CI=1.23– 5.85; p< 0.01) were statistically associated. In total, 98.4% of enteric bacterial isolates were sensitive for Meropenem, whereas 82.5% were resistant against Ampicillin. Multidrug resistance was detected in 49.2% of enteric bacteria.Conclusion: We found that enteric bacteria are common causative agents of diarrhea in immune-compromised patients. The high rate of drug resistance calls for escalating antimicrobial susceptibility testing before prescribing antimicrobial agent.Keywords: enteric pathogen, human immunodeficiency virus, diarrhea

Vibrio cholerae is a non-invasive enteric pathogen known to cause a major public health problem called cholera. The pathogen inhabits the aquatic environment while outside the human host, it is transmitted into the host easily through ingesting contaminated food and water containing the vibrios, thus causing diarrhoea and vomiting. V. cholerae must resist several layers of colonization resistance mechanisms derived from the host or the gut commensals to successfully survive, grow, and colonize the distal intestinal epithelium, thus causing an infection. The colonization resistance mechanisms derived from the host are not specific to V. cholerae but to all invading pathogens. However, some of the gut commensal-derived colonization resistance may be more specific to the pathogen, making it more challenging to overcome. Consequently, the pathogen has evolved well-coordinated mechanisms that sense and utilize the anti-colonization factors to modulate events that promote its survival and colonization in the gut. This review is aimed at discussing how V. cholerae interacts and resists both host- and microbe-specific colonization resistance mechanisms to cause infection.

Human excreta can be used as biofertilizers due to their nutrient and organic matter content. Nevertheless, the behavior of microorganisms should be investigated, as enteric pathogens can accumulate in the soil. Therefore, we evaluated the survival and transport of two enteric bacteria (E. coli and Salmonella enterica) and two enteric virus surrogates (MS2 and ΦX-174 bacteriophages) when applied as contaminants of four biofertilizers on Lactuca sativa production. The study was carried out in lysimeters outdoors with repacked silt loam soil, using a randomized block design (RBD) with six treatments and three replicates. Biofertilizers were urea-treated feces, feces composted with organic waste, stored liquid urine, and struvite and were inoculated at 106–107 cfu/pfu g−1 of E. coli and bacteriophages strains and 107–109 of S. enterica. Soil amended with composted feces exhibited the lowest decimal reductions (T90) for the microorganisms evaluated, with 10.2 days for E. coli, 11.9 days for S. enterica, 22.2 days for MS2, and 3.7 days for ΦX-174. Urea-treated feces temporarily hindered the growth and survival of E. coli and S. enterica in the soil. However, both bacteria were present after one month, while MS2 showed a stable concentration with this biofertilizer. E. coli presented a stable behavior when applied with urine, whereas MS2 was stable with urea-treated feces, urine, and struvite. ΦX-174 and S. enterica were not considered good representatives of most resistant enteric pathogens’ behavior in the soil during food production since they were not detected in leachate and showed a rapid die-off in soil. E. coli leached faster and in higher concentrations than MS2, which presented higher concentrations in deeper soil layers, from 10 to 20 cm. Therefore, using urine, struvite, and urea-treated faces implied a higher chance of infection, as a decimal reduction time was not observed during the 36 days of the experiment. As a result, in short-term cultures, it is recommended that additional hygiene barriers are taken to avoid infection since E. coli and MS2 were still detected at the end of the experiment. An interval of 90 days between fertilizing and harvesting is recommended to prevent pathogen infection. These results are the basis for a quantitative risk assessment of human excreta-based biofertilizers for food production.

Vibrio cholerae is a non-invasive enteric pathogen known to cause a major public health problem called cholera. The pathogen inhabits the aquatic environment while outside the human host, it is transmitted into the host easily through ingesting contaminated food and water containing the vibrios, thus causing diarrhoea and vomiting. V. cholerae must resist several layers of colonization resistance mechanisms derived from the host or the gut commensals to successfully survive, grow, and colonize the distal intestinal epithelium, thus causing an infection. The colonization resistance mechanisms derived from the host are not specific to V. cholerae but to all invading pathogens. However, some of the gut commensal-derived colonization resistance may be more specific to the pathogen, making it more challenging to overcome. Consequently, the pathogen has evolved wellcoordinated mechanisms that sense and utilize the anti-colonization factors to modulate events that promote its survival and colonization in the gut. This review is aimed at discussing how V. cholerae interacts and resists both host- and microbe-specific colonization resistance mechanisms to cause infection. [ABSTRACT FROM AUTHOR]

Shigellosis is a serious foodborne diarrheal disease caused by the Shigella species. It is a critical global health issue. In developing countries, shigellosis causes most of the mortality in children below 5 years of age. Globally, around 165 million cases of diarrhea caused by Shigella are reported, which accounts for almost 1 million deaths, in which the majority are recorded in Third World nations. In this study, silver nanoparticles were synthesized using Mangifera indica kernel (MK-AgNPs) seed extracts. The biosynthesized M. indica silver nanoparticles (MK-AgNPs) were characterized using an array of spectroscopic and microscopic tools, such as UV–Vis, scanning electron microscopy, particle size analyzer, Fourier transform infrared spectroscopy, and X-ray diffractometer. The nanoparticles were spherical in shape and the average size was found to be 42.7 nm. The MK-AgNPs exhibited remarkable antibacterial activity against antibiotic-resistant clinical Shigella sp. The minimum inhibitory concentration (MIC) value of the MK-AgNPs was found to be 20 μg/mL against the multi-drug-resistant strain Shigella flexneri. The results clearly demonstrate that MK-AgNPs prepared using M. indica kernel seed extract exhibited significant bactericidal action against pathogenic Shigella species. The biosynthesized nanoparticles from mango kernel could possibly prove therapeutically useful and effective in combating the threat of shigellosis after careful investigation of its toxicity and in vivo efficacy. [ABSTRACT FROM AUTHOR]

Every year, enteric infections and associated diarrhea kill millions of people. The situation is compounded by increases in the number of enteric pathogens that are acquiring resistance to antibiotics, as well as (hitherto) a relative paucity of information on host molecular targets that may contribute to diarrhea. Many forms of diarrheal disease depend on the dysregulation of intestinal ion transporters, and an associated imbalance between secretory and absorptive functions of the intestinal epithelium. A number of major transporters have been implicated in the pathogenesis of diarrheal diseases and thus an understanding of their expression, localization, and regulation after infection with various bacteria, viruses, and protozoa likely will prove critical in designing new therapies. This article surveys our understanding of transporters that are modulated by specific pathogens and the mechanism(s) involved, thereby illuminating targets that might be exploited for new therapeutic approaches.

ABSTRACT Salmonella enterica is one of the most common foodborne pathogens and, due to the spread of antibiotic resistance, new antimicrobial strategies are urgently needed to control it. In this study, we explored the probiotic potential of Bacillus subtilis PS-216 and elucidated the mechanisms that underlie the interactions between this soil isolate and the model pathogenic strain S. Typhimurium SL1344. The results reveal that B. subtilis PS-216 inhibits the growth and biofilm formation of S. Typhimurium through the production of the pks cluster-dependent polyketide bacillaene. The presence of S. Typhimurium enhanced the activity of the PpksC promoter that controls bacillaene production, suggesting that B. subtilis senses and responds to Salmonella. The level of Salmonella inhibition, overall PpksC activity, and PpksC induction by Salmonella were all higher in nutrient-rich conditions than in nutrient-depleted conditions. Although eliminating the extracellular polysaccharide production of B. subtilis via deletion of the epsA-O operon had no significant effect on inhibitory activity against Salmonella in nutrient-rich conditions, this deletion mutant showed an enhanced antagonism against Salmonella in nutrient-depleted conditions, revealing an intricate relationship between exopolysaccharide production, nutrient availability, and bacillaene synthesis. Overall, this work provides evidence on the regulatory role of nutrient availability, sensing of the competitor, and EpsA-O polysaccharide in the social outcome of bacillaene-dependent competition between B. subtilis and S. Typhimurium. IMPORTANCE Probiotic bacteria represent an alternative for controlling foodborne disease caused by Salmonella enterica, which constitutes a serious concern during food production due to its antibiotic resistance and resilience to environmental stress. Bacillus subtilis is gaining popularity as a probiotic, but its behavior in biofilms with pathogens such as Salmonella remains to be elucidated. Here, we show that the antagonism of B. subtilis is mediated by the polyketide bacillaene and that the production of bacillaene is a highly dynamic trait which depends on environmental factors such as nutrient availability and the presence of competitors. Moreover, the production of extracellular polysaccharides by B. subtilis further alters the influence of these factors. Hence, this work highlights the inhibitory effect of B. subtilis, which is condition-dependent, and the importance of evaluating probiotic strains under conditions relevant to the intended use.

ABSTRACT The application of clinical diagnostics for gastroenteritis in children has implications for a broad collection of stakeholders, impacting clinical care, communicable disease control, and laboratory utilization. To support diagnostic stewardship as gastroenteritis testing options continue to advance, it is critical to understand which enteropathogens constitute priorities for testing across stakeholder groups. Using a modified Delphi technique, we elicited opinions of subject matter experts to determine clinical and public health testing priorities. There was a high level of overall agreement (≥80%) among stakeholders (final round n = 15) that testing was important for Campylobacter, Escherichia coli O157 and other Shiga toxin-producing E. coli, Salmonella, Shigella, Vibrio, Yersinia, norovirus, and rotavirus. Immunocompromised children were identified as a special population that warranted the additional testing of three to four bacterial and parasitic targets. To support these clinical and public health testing priorities, diagnostic stewardship strategies can be employed, such as educating clinicians, developing new decision support tools, and using multiplex testing in concert with selective result reporting and annotation. IMPORTANCE Children with diarrhea and vomiting who seek care can be infected with a wide variety of infectious agents. This study reports findings from a survey of clinical, public health, and laboratory subject matter experts on the infectious agents that are most important to test for. The majority agreed on the importance of testing children likely infected with several bacterial agents, as well as two common viruses. Although confirming a child is positive for a viral agent is unlikely to change clinical care, participants noted the importance of monitoring these viruses for public health purposes. To avoid over-testing children, however, these results should be used to support diagnostic stewardship strategies and design new decision support tools.

We compared the Allplex Gastrointestinal V/B1/B2 Assays and Seeplex Diarrhea V/B1/B2 ACE Detection Assays in patients with acute gastroenteritis (AGE). Of the total 432 specimens, 48.8% and 54.9% samples were positive for any bacterial or viral target using Seeplex and Allplex, respectively (P = 0.002). The overall percent agreement (OPA) between the two panels was >95% and the lowest OPA was 95.4% for CdB. Allplex identified 40 samples positive for Salmonella spp. , while Seeplex and OBC identified only 27 (67.5%) and 8 (20%), respectively. Shigella spp. were detected by assays in six samples, but none were identified using culture. Clostridium perfringens with Seeplex was detected in 70 (16.2%). It remained an informative species in identifying AGE although cpe gene showed only 9.8% positivity. Pathogenic Escherichia coli with Allplex could be detected in 40 (9.3%) samples, which could provide valuable information for the diagnosis of AGE. [ABSTRACT FROM AUTHOR]

Shigellosis is a serious foodborne diarrheal disease caused by the Shigella species. It is a critical global health issue. In developing countries, shigellosis causes most of the mortality in children below 5 years of age. Globally, around 165 million cases of diarrhea caused by Shigella are reported, which accounts for almost 1 million deaths, in which the majority are recorded in Third World nations. In this study, silver nanoparticles were synthesized using Mangifera indica kernel (MK-AgNPs) seed extracts. The biosynthesized M. indica silver nanoparticles (MK-AgNPs) were characterized using an array of spectroscopic and microscopic tools, such as UV–Vis, scanning electron microscopy, particle size analyzer, Fourier transform infrared spectroscopy, and X-ray diffractometer. The nanoparticles were spherical in shape and the average size was found to be 42.7 nm. The MK-AgNPs exhibited remarkable antibacterial activity against antibiotic-resistant clinical Shigella sp. The minimum inhibitory concentration (MIC) value of the MK-AgNPs was found to be 20 μg/mL against the multi-drug-resistant strain Shigella flexneri. The results clearly demonstrate that MK-AgNPs prepared using M. indica kernel seed extract exhibited significant bactericidal action against pathogenic Shigella species. The biosynthesized nanoparticles from mango kernel could possibly prove therapeutically useful and effective in combating the threat of shigellosis after careful investigation of its toxicity and in vivo efficacy.

Prior to the implementation of new diagnostic techniques, a thorough evaluation is mandatory in order to ensure diagnostic reliability. If positive samples are scarcely available, however, such evaluations can be difficult to perform. Here, we evaluated four SeeGene Allplex real-time PCR assays amplifying a total of 28 bacteria, microsporidal and parasitic nucleic acid sequence targets in human stool samples in a multicentric approach. In the assessments with strongly positive samples, sensitivity values ranging between 13% and 100% were recorded for bacteria, between 0% and 100% for protozoa and between 7% and 100% for helminths and microsporidia; for the weakly positive samples, the recorded sensitivity values for bacteria ranged from 0% to 100%; for protozoa, from 0% to 40%; and for helminths and microsporidia, from 0% to 53%. For bacteria, the recorded specificity was in the range between 87% and 100%, while a specificity of 100% was recorded for all assessed PCRs targeting parasites and microsporidia. The intra- and inter-assay variations were generally low. Specifically for some helminth species, the sensitivity could be drastically increased by applying manual nucleic acid extraction instead of the manufacturer-recommended automatic procedure, while such effects were less obvious for the bacteria and protozoa. In summary, the testing with the chosen positive control samples showed varying degrees of discordance between the evaluated Allplex assays and the applied in-house reference assays associated with higher cycle threshold values in the Allplex assays, suggesting that samples with very low pathogen densities might be missed. As the targeted species can occur as harmless colonizers in the gut of individuals in high-endemicity settings as well, future studies should aim at assessing the clinical relevance of the latter hint. [ABSTRACT FROM AUTHOR]

Background: Lettuce is linked to recurrent outbreaks of Shiga toxin-producing Escherichia coli (STEC) infections, the seasonality of which remains unresolved. Infections have occurred largely from processed lettuce, which undergoes substantial physiological changes during storage. We investigated the microbiome and STEC O157:H7 (EcO157) colonization of fresh-cut lettuce of two cultivars with long and short shelf life harvested in the spring and fall in California and stored in modified atmosphere packaging (MAP) at cold and warm temperatures. Results: Inoculated EcO157 declined significantly less on the cold-stored cultivar with short shelf life, while multiplying rapidly at 24 °C independently of cultivar. Metagenomic sequencing of the lettuce microbiome revealed that the pre-storage bacterial community was variable but dominated by species in the Erwiniaceae and Pseudomonadaceae. After cold storage, the microbiome composition differed between cultivars, with a greater relative abundance (RA) of Erwiniaceae and Yersiniaceae on the cultivar with short shelf life. Storage at 24 °C shifted the microbiome to higher RAs of Erwiniaceae and Enterobacteriaceae and lower RA of Pseudomonadaceae compared with 6 °C. Fall harvest followed by lettuce deterioration were identified by recursive partitioning as important factors associated with high EcO157 survival at 6 °C, whereas elevated package CO2 levels correlated with high EcO157 multiplication at 24 °C. EcO157 population change correlated with the lettuce microbiome during 6 °C storage, with fall microbiomes supporting the greatest EcO157 survival on both cultivars. Fall and spring microbiomes differed before and during storage at both temperatures. High representation of Pantoea agglomerans was a predictor of fall microbiomes, lettuce deterioration, and enhanced EcO157 survival at 6 °C. In contrast, higher RAs of Erwinia persicina, Rahnella aquatilis, and Serratia liquefaciens were biomarkers of spring microbiomes and lower EcO157 survival. Conclusions: The microbiome of processed MAP lettuce evolves extensively during storage. Under temperature abuse, high CO2 promotes a lettuce microbiome enriched in taxa with anaerobic capability and EcO157 multiplication. In cold storage, our results strongly support a role for season and lettuce deterioration in EcO157 survival and microbiome composition, suggesting that the physiology and microbiomes of fall- and spring-harvested lettuce may contribute to the seasonality of STEC outbreaks associated with lettuce grown in coastal California. [ABSTRACT FROM AUTHOR]

Mitigation strategies to prevent microbial contamination of crops are lacking. We tested the hypothesis that induction of plant systemic resistance by biological (induced systemic resistance [ISR]) and chemical (systemic acquired resistance [SAR]) elicitors reduces endophytic colonization of leaves by Salmonella enterica serovars Senftenberg and Typhimurium. S. Senftenberg had greater endophytic fitness than S. Typhimurium in basil and lettuce. The apoplastic population sizes of serovars Senftenberg and Typhimurium in basil and lettuce, respectively, were significantly reduced approximately 10- to 100-fold by root treatment with microbial inducers of systemic resistance compared to H2O treatment. Rhodotorula glutinis effected the lowest population increases of S. Typhimurium in lettuce and S. Senftenberg in basil leaves, respectively 120- and 60-fold lower than those seen with the H2O treatment over 10 days postinoculation. Trichoderma harzianum and Pichia guilliermondii did not have any significant effect on S. Senftenberg in the basil apoplast. The chemical elicitors acidobenzolar-S-methyl and DL-b-amino-butyric acid inhibited S. Typhimurium multiplication in the lettuce apoplast 10- and 2-fold, respectively, compared to H2O-treated plants. All ISR and SAR inducers applied to lettuce roots in this study increased leaf expression of the defense gene PR1, as did Salmonella apoplastic colonization in H2O-treated lettuce plants. Remarkably, both acidobenzolar-S-methyl upregulation and R. glutinis upregulation of PR1 were repressed by the presence of Salmonella in the leaves. However, enhanced PR1 expression was sustained longer and at greater levels upon elicitor treatment than by Salmonella induction alone. These results serve as a proof of concept that priming of plant immunity may provide an intrinsic hurdle against the endophytic establishment of enteric pathogens in leafy vegetables. [ABSTRACT FROM AUTHOR]

Outbreaks of produce-associated foodborne illness continue to pose a threat to human health worldwide. New approaches are necessary to improve produce safety. Plant innate immunity has potential as a host-based strategy for the deactivation of enteric pathogens. In response to various biotic and abiotic threats, plants mount defense responses that are governed by signaling pathways. Once activated, these result in the release of reactive oxygen and nitrogen species in addition to secondary metabolites that aim at tempering microbial infection and pest attack. These phytochemicals have been investigated as alternatives to chemical sanitization, as many are effective antimicrobial compounds in vitro. Their antagonistic activity toward enteric pathogens may also provide an intrinsic hurdle to their viability and multiplication in planta. Plants can detect and mount basal defenses against enteric pathogens. Evidence supports the role of plant bioactive compounds in the physiology of Salmonella enterica, Escherichia coli, and Listeria monocytogenes as well as their fitness on plants. Here, we review the current state of knowledge of the effect of phytochemicals on enteric pathogens and their colonization of plants. Further understanding of the interplay between foodborne pathogens and the chemical environment on/in host plants may have lasting impacts on crop management for enhanced microbial safety through translational applications in plant breeding, editing technologies, and defense priming. [ABSTRACT FROM AUTHOR]

Abstract Background The search for alternatives to antibiotics in pig production has increased the interest in natural resources with antimicrobial properties, such as medium-chain fatty acids (MCFA) as in-feed additives. This study evaluated the potential of a novel blend of MCFA salts (DIC) from distilled coconut oil with a lauric acid content to reduce enteropathogens and control intestinal diseases around weaning. Two experimental disease models were implemented in early-weaned piglets, consisting of two oral challenges: Salmonella Typhimurium (1.2 × 108 CFU) or enterotoxigenic Escherichia coli (ETEC) F4 (1.5 × 109 CFU). The parameters assessed were: animal performance, clinical signs, pathogen excretion, intestinal fermentation, immune-inflammatory response, and intestinal morphology. Results The Salmonella challenge promoted an acute course of diarrhea, with most of the parameters responding to the challenge, whereas the ETEC F4 challenge promoted a mild clinical course. A consistent antipathogenic effect of DIC was observed in both trials in the hindgut, with reductions in Salmonella spp. plate counts in the cecum (P = 0.03) on d 8 post-inoculation (PI) (Salmonella trial), and of enterobacteria and total coliform counts in the ileum and colon (P < 0.10) on d 8 PI (ETEC F4 trial). When analyzing the entire colonic microbiota (16S rRNA gene sequencing), this additive tended (P = 0.13) to reduce the Firmicutes/Bacteroidetes ratio and enriched Fibrobacteres after the Salmonella challenge. In the ETEC F4 challenge, DIC prompted structural changes in the ecosystem with increases in Dialister, and a trend (P = 0.14) to increase the Veillonellaceae family. Other parameters such as the intestinal fermentation products or serum pro-inflammatory mediators were not modified by DIC supplementation, nor were the histological parameters. Only the intraepithelial lymphocyte (IEL) counts were lowered by DIC in animals challenged with Salmonella (P = 0.07). With ETEC F4, the IEL counts were higher with DIC on d 8 PI (P = 0.08). Conclusions This study confirms the potential activity of this MCFA salts mixture to reduce intestinal colonization by opportunistic pathogens such as Salmonella or E. coli and its ability to modulate colonic microbiota. These changes could explain to some extent the local immune cell response at the ileal level.

1. This review describes different classes of antimicrobial peptides (AMP) found in the gastrointestinal (GI) tract of avian species, and their antimicrobial and immunomodulatory activities. The potential benefits of synthetic AMP in poultry production are examined, in the context of the use of AMP as alternatives to antimicrobial growth promoters (AGP). 2. Since the mid-1950s, antibiotic growth promoters (AGP) have been used in feed at low prophylactic doses to modulate the homoeostasis of intestinal microbiota, decreasing the risk of intestinal dysbacteriosis and the growth of pathogens within the avian gut. Over the last three decades, AGP have faced major regulatory restrictions due to concerns of generating antimicrobial resistance (AMR). It is now well documented that the rate of infectious disease outbreaks is higher in flocks that are not fed prophylactic antibiotics, resulting in a compensatory increase in antimicrobial use for therapeutic purposes. 3. Endogenous natural AMP production is associated with the presence of microbiota and their interaction with the intestinal epithelial and lamina propria lymphoid cells. Their antimicrobial activity shapes the beneficial microbiota population and controls intestinal pathogens such Clostridium and Salmonella spp., and stimulates the development and maturation of the local immune system. 4. Similar to AGP, AMP can establish a well-balanced gut beneficial microbiota for adequate immune-competence, animal health and high growth performance parameters such as feed intake, daily weight, feed conversion and accumulated mortality. 5. Antimicrobial proteins and peptides constitute an essential part of the innate immune system of all organisms and protect the host from invading pathogenic bacteria, viruses, fungi, and parasites by interacting with the negatively charged pathogen membranes. [ABSTRACT FROM AUTHOR]

Heat resistance, D- and z-values of five Salmonella enterica strains, namely S. Senftenberg LTH 5703, S. Typhimurium ATCC 13311 and ATCC 14028, S. Saintpaul LTH 6494, and S. Enteritidis ATCC 13076, as well as of a Salmonella cocktail and three potential surrogate strains were investigated in phosphate buffered saline and strawberry nectar (12° Brix). Thermal inactivation was performed at 60 °C, 65 °C, and 72 °C. Moreover, the influence of pre-incubation under stressful environmental conditions on the heat resistance was tested. The results identified Escherichia coli ATCC 11229 as a suitable surrogate candidate for Salmonella strains, as its calculated D-values in both matrices were higher or statistically the same than those of the examined pathogens. The safety analysis showed no virulence factors that could classify E. coli ATCC 11229 as a pathogen. Furthermore, the results show significant differences in D-values of all strains in different matrices, which indicates a clear influence of the environment on heat resistance of bacteria. The pre-incubation under stress conditions had no significant effect on the heat resistance of E. coli ATCC 11229. However, further research is needed to explore the influence of stress conditions and duration on bacterial heat resistance. • E. coli ATCC 11229 is a suitable surrogate for selected Salmonella strains. • Thermal resistance of Salmonella cocktail corresponds to that of individual strains. • Bacterial heat resistance differs in strawberry nectar and neutral environment. • Pre-incubation of E. coli ATCC 11229 under stress does not induce cross-protection. [ABSTRACT FROM AUTHOR]

Enterotoxigenic Escherichia coli (ETEC) is a major cause of travelers’ diarrhea and of diarrhea among young children in developing countries. Experimental challenge studies in adult volunteers have played a pivotal role in establishing ETEC as an enteric pathogen, elucidating its pathogenesis by identifying specific virulence attributes, characterizing the human immune response to clinical and sub-clinical ETEC infection and assessing preliminarily the clinical acceptability, immunogenicity and efficacy of prototype ETEC vaccines. This review provides a historical perspective of experimental challenge studies with ETEC. It summarizes pioneering early studies carried out by investigators at the University of Maryland School of Medicine to show how those studies provided key information that influenced the directions taken by many research groups to develop vaccines to prevent ETEC. In addition, key experimental challenge studies undertaken at other institutions will also be cited.

Although rodents are well-known reservoirs and vectors for a number of zoonoses, the functional role that peridomestic rodents serve in the amplification and transmission of foodborne pathogens is likely underappreciated. Clear links have been identified between commensal rodents and outbreaks of foodborne pathogens throughout Europe and Asia; however, comparatively little research has been devoted to studying this relationship in the United States. In particular, regional studies focused on specific rodent species and their foodborne pathogen reservoir status across the diverse agricultural landscapes of the United States are lacking. We posit that both native and invasive species of rodents associated with food-production pipelines are likely sources of seasonal outbreaks of foodborne pathogens throughout the United States. In this study, we review the evidence that identifies peridomestic rodents as reservoirs for foodborne pathogens, and we call for novel research focused on the metagenomic communities residing at the rodent-agriculture interface. Such data will likely result in the identification of new reservoirs for foodborne pathogens and species-specific demographic traits that might underlie seasonal enteric disease outbreaks. Moreover, we anticipate that a One Health metagenomic research approach will result in the discovery of new strains of zoonotic pathogens circulating in peridomestic rodents. Data resulting from such research efforts would directly inform and improve upon biosecurity efforts, ultimately serving to protect our food supply. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is one of the largest threats to global health and imposes substantial burdens in terms of morbidity, mortality, and economic costs. The gut is a key conduit for the genesis and spread of antimicrobial resistance in enteric bacterial pathogens. Distinct bacterial species that cause enteric disease can exist as invasive enteropathogens that immediately evoke gastrointestinal distress, or pathobionts that can arise from established bacterial commensals to inflict dysbiosis and disease. Furthermore, various environmental reservoirs and stressors facilitate the evolution and transmission of resistance. In this review, we present a comprehensive discussion on circulating resistance profiles and gene mobilization strategies of the most problematic species of enteric bacterial pathogens. Importantly, we present emerging approaches toward surveillance of pathogens and their resistance elements as well as promising treatment strategies that can circumvent common resistance mechanisms.

Prior to the implementation of new diagnostic techniques, a thorough evaluation is mandatory in order to ensure diagnostic reliability. If positive samples are scarcely available, however, such evaluations can be difficult to perform. Here, we evaluated four SeeGene Allplex real-time PCR assays amplifying a total of 28 bacteria, microsporidal and parasitic nucleic acid sequence targets in human stool samples in a multicentric approach. In the assessments with strongly positive samples, sensitivity values ranging between 13% and 100% were recorded for bacteria, between 0% and 100% for protozoa and between 7% and 100% for helminths and microsporidia; for the weakly positive samples, the recorded sensitivity values for bacteria ranged from 0% to 100%; for protozoa, from 0% to 40%; and for helminths and microsporidia, from 0% to 53%. For bacteria, the recorded specificity was in the range between 87% and 100%, while a specificity of 100% was recorded for all assessed PCRs targeting parasites and microsporidia. The intra- and inter-assay variations were generally low. Specifically for some helminth species, the sensitivity could be drastically increased by applying manual nucleic acid extraction instead of the manufacturer-recommended automatic procedure, while such effects were less obvious for the bacteria and protozoa. In summary, the testing with the chosen positive control samples showed varying degrees of discordance between the evaluated Allplex assays and the applied in-house reference assays associated with higher cycle threshold values in the Allplex assays, suggesting that samples with very low pathogen densities might be missed. As the targeted species can occur as harmless colonizers in the gut of individuals in high-endemicity settings as well, future studies should aim at assessing the clinical relevance of the latter hint.

Background: Clostridium difficile infection (CDI) is one of the most common hospital-acquired infections and is associated with significant morbidity and mortality. Since owning a cat or dog could enrich the gut microbiome, we hypothesized that it would be protective against CDI. Aims: We conducted a survey study on patients tested for CDI in order to assess whether living in the presence of a pet is associated with a decreased risk of CDI. Methods: We surveyed subjects aged 18–90 over a 14-month period using a retrospective case–control design. Subjects with CDI were matched by gender and age to patients who tested negative and had no prior history of CDI. A web-based survey was provided to subjects by mail or assisted by phone. Conditional logistic regression was used to assess for associations between CDI and the various risk factors. Results: 205 CDI positive and 205 CDI negative subjects (response rate of 50.2%) were included. After matching for age and sex, living with a cat or dog was not associated with negative CDI testing. Exploratory multivariable modeling identified an unexpected association between positive CDI testing and high meat intake (OR 2.13, 95% CI 1.21–3.77) as well as between positive CDI testing and cat allergies (OR 1.88, 95% CI 1.02–3.46). Conclusion: Living with a cat or dog was not associated with negative CDI testing. Several novel risk factors for CDI have been identified including high meat intake and cat allergies. [ABSTRACT FROM AUTHOR]

ABSTRACT Cryptosporidium parvum and Cryptosporidium hominis have emerged as major enteric pathogens of infants in the developing world, in addition to their known importance in immunocompromised adults. Although there has been recent progress in identifying new small molecules that inhibit Cryptosporidium sp. growth in vitro or in animal models, we lack information about their mechanism of action, potency across the life cycle, and cidal versus static activities. Here, we explored four potent classes of compounds that include inhibitors that likely target phosphatidylinositol 4 kinase (PI4K), phenylalanine-tRNA synthetase (PheRS), and several potent inhibitors with unknown mechanisms of action. We utilized monoclonal antibodies and gene expression probes for staging life cycle development to define the timing of when inhibitors were active during the life cycle of Cryptosporidium parvum grown in vitro. These different classes of inhibitors targeted different stages of the life cycle, including compounds that blocked replication (PheRS inhibitors), prevented the segmentation of daughter cells and thus blocked egress (PI4K inhibitors), or affected sexual-stage development (a piperazine compound of unknown mechanism). Long-term cultivation of C. parvum in epithelial cell monolayers derived from intestinal stem cells was used to distinguish between cidal and static activities based on the ability of parasites to recover from treatment. Collectively, these approaches should aid in identifying mechanisms of action and for designing in vivo efficacy studies based on time-dependent concentrations needed to achieve cidal activity. IMPORTANCE Currently, nitazoxanide is the only FDA-approved treatment for cryptosporidiosis; unfortunately, it is ineffective in immunocompromised patients, has varied efficacy in immunocompetent individuals, and is not approved in infants under 1 year of age. Identifying new inhibitors for the treatment of cryptosporidiosis requires standardized and quantifiable in vitro assays for assessing potency, selectivity, timing of activity, and reversibility. Here, we provide new protocols for defining which stages of the life cycle are susceptible to four highly active compound classes that likely inhibit different targets in the parasite. We also utilize a newly developed long-term culture system to define assays for monitoring reversibility as a means of defining cidal activity as a function of concentration and time of treatment. These assays should provide valuable in vitro parameters to establish conditions for efficacious in vivo treatment.

Every year, enteric infections and associated diarrhea kill millions of people. The situation is compounded by increases in the number of enteric pathogens that are acquiring resistance to antibiotics, as well as (hitherto) a relative paucity of information on host molecular targets that may contribute to diarrhea. Many forms of diarrheal disease depend on the dysregulation of intestinal ion transporters, and an associated imbalance between secretory and absorptive functions of the intestinal epithelium. A number of major transporters have been implicated in the pathogenesis of diarrheal diseases and thus an understanding of their expression, localization, and regulation after infection with various bacteria, viruses, and protozoa likely will prove critical in designing new therapies. This article surveys our understanding of transporters that are modulated by specific pathogens and the mechanism(s) involved, thereby illuminating targets that might be exploited for new therapeutic approaches.

Outer membrane proteins are commonly produced by gram-negative bacteria, and they have diverse functions. A subgroup of proteins, which includes OmpA, OmpW and OmpX, is often involved in bacterial pathogenesis. Here we show that OmpA, rather than OmpW or OmpX, contributes to the virulence of enterohemorrhagic Escherichia coli (EHEC) through its type III secretion system (T3SS). Deletion of ompA decreased secretion of the T3SS proteins EspA and EspB; however, the expression level of the LEE genes that encode a set of T3SS proteins did not decrease. The ompA mutant had less abilities to form A/E lesions in host epithelial cells and lyse human red blood cells than the parent strain. Moreover, the virulence of an ompA mutant of Citrobacter rodentium (traditionally used to estimate T3SS-associated virulence in mice) was attenuated. Mice infected with the ompA mutant survived longer than those infected with the parent strain. Furthermore, mice infected with ompA developed symptoms of diarrhea more slowly than mice infected with the parent strain. Altogether, these results suggest that OmpA sustains the activity of the T3SS and is required for optimal virulence in EHEC. This work expands the roles of outer membrane proteins in bacterial pathogenesis.

Clostridioides difficile is a spore-forming enteric pathogen causing life-threatening diarrhoea and colitis. Microbial disruption caused by antibiotics has been linked with susceptibility to, and transmission and relapse of, C. difficile infection. Therefore, there is an urgent need for novel therapeutics that are effective in preventing C. difficile growth, spore germination, and outgrowth. In recent years bacteriophage-derived endolysins and their derivatives show promise as a novel class of antibacterial agents. In this study, we recombinantly expressed and characterized a cell wall hydrolase (CWH) lysin from C. difficile phage, phiMMP01. The full-length CWH displayed lytic activity against selected C. difficile strains. However, removing the N-terminal cell wall binding domain, creating CWH351—656, resulted in increased and/or an expanded lytic spectrum of activity. C. difficile specificity was retained versus commensal clostridia and other bacterial species. As expected, the putative cell wall binding domain, CWH1—350, was completely inactive. We also observe the effect of CWH351—656 on preventing C. difficile spore outgrowth. Our results suggest that CWH351—656 has therapeutic potential as an antimicrobial agent against C. difficile infection.

Abstract Background The Gram-negative bacterium Vibrio vulnificus can cause severe disease in humans who consume undercooked, contaminated seafood. To study food-borne V. vulnificus disease in the laboratory, mouse virulence studies predominantly use death as the primary experimental endpoint because behaviorally based moribund status does not consistently predict lethality. This study assessed ventral surface temperature (VST) and its association with mouse survival during V. vulnificus virulence studies as an efficacious, humane alternative. Methods VST of mice intragastrically inoculated with V. vulnificus was measured every 2-h for 24 h and data for minimal VST analyzed for prediction of lethal outcome. Results In contrast to the relatively stable VST of mock-infected control animals, mice infected with V. vulnificus exhibited hypothermia with minima occurring 8 to 12 h post-inoculation. The minimum VST of mice that proceeded to death was significantly lower than that of surviving mice. VST ≤ 23.5 °C was predictive of subsequent death with a sensitivity of 68% and specificity of 95%. Conclusions Use of VST ≤ 23.5 °C as an experimental endpoint during V. vulnificus infection has potential to reduce suffering of nearly 70% of mice for a mean of 10 h per mouse, without compromising experimental efficacy. Temperature cutoff of 23.5 °C exhibited 93% positive and 77% negative predictive value. For future V. vulnificus virulence studies requiring only binary comparison (e.g., LD50 assays), we find that VST can be applied as a humane endpoint. However, use of VST is not recommended when detailed survival kinetics are desired.

Abstract Background Tritrichomonas foetus is an emergent and important enteric pathogen of cats, which causes prolonged diarrhoea in cats. Case presentation This study describes a T. foetus infection in a seven-month-old, entire male domestic shorthair kitten with a six-month history of persistent large intestinal diarrhoea, faecal incontinence, prostration, apathy and weight loss. Parasites were microscopically observed and confirmed by PCR and DNA sequencing. Molecular analyses were carried out comparing the sequence obtained in this study with T. foetus and T. suis. Retrieved from GenBank. After treatment with ronidazole, the cat showed resolution of clinical signs. Conclusions This is the first clinical case of T. foetus infection in a chronic diarrheic cat in Brazil and South America, confirming the presence of this pathogen in this part of the world and highlighting the importance of this protozoa being considered in the differential diagnosis of cats presenting diarrhoea of the large intestine. Our case report enriches our knowledge on the geographical distribution of T. foetus in cats in Brazil and provides further understanding of the clinical significance of feline intestinal trichomoniasis in this country.

The contact-dependent type VI secretion system (T6SS) in diverse microbes plays crucial roles in both inter-bacterial and bacteria-host interactions. As numerous microorganisms inhabit the intestinal ecosystem at a high density, it is necessary to consider the functions of T6SS in intestinal bacteria. In this mini-review, we discuss T6SS-dependent functions in intestinal microbes, including commensal microbes and enteric pathogens, and list experimentally verified species of intestinal bacteria containing T6SS clusters. Several seminal studies have shown that T6SS plays crucial antibacterial roles in colonization resistance, niche occupancy, activation of host innate immune responses, and modulation of host intestinal mechanics. Some potential roles of T6SS in the intestinal ecosystem, such as targeting of single cell eukaryotic competitors, competition for micronutrients, and stress resistance are also discussed. Considering the distinct activities of T6SS in diverse bacteria residing in the intestine, we suggest that T6SS research in intestinal microbes may be beneficial for the future development of new medicines and clinical treatments.

Background: In sub-Saharan Africa, molecular epidemiological investigation of outbreaks caused by antimicrobial-resistant enteric bacterial pathogens have mostly been described for Salmonella species, Vibrio cholerae, Shigella species and Escherichia coli. For these organisms, I reviewed all publications describing the use of molecular subtyping methodologies to investigate outbreaks caused by multidrug-resistant (MDR) enteric bacterial infections. Objectives: To describe the use of molecular subtyping methodologies to investigate outbreaks caused by MDR enteric bacterial pathogens in sub-Saharan Africa and to describe the current status of molecular subtyping capabilities in the region. Methods: A PubMed database literature search (English language only) was performed using the search strings: ‘Africa outbreak MDR’, ‘Africa outbreak multi’, ‘Africa outbreak multidrug’, ‘Africa outbreak multi drug’, ‘Africa outbreak resistance’, ‘Africa outbreak resistant’, ‘Africa outbreak drug’, ‘Africa outbreak antibiotic’, ‘Africa outbreak antimicrobial’. These search strings were used in combination with genus and species names of the organisms listed above. All results were included in the review. Results: The year 1991 saw one of the first reports describing the use of molecular subtyping methodologies in sub-Saharan Africa; this included the use of plasmid profiling to characterise Salmonella Enteritidis. To date, several methodologies have been used; pulsed-field gel electrophoresis analysis and multilocus sequence typing have been the most commonly used methodologies. Investigations have particularly highlighted the emergence and spread of MDR clones; these include Salmonella Typhi H58 and Salmonella Typhimurium ST313 clones. In recent times, whole-genome sequencing (WGS) analysis approaches have increasingly been used. Conclusion: Traditional molecular subtyping methodologies are still commonly used and still have their place in investigations; however, WGS approaches have increasingly been used and are slowly gaining a stronghold. African laboratories need to start adapting their molecular surveillance methodologies to include WGS, as it is foreseen that WGS analysis will eventually replace all traditional methodologies.

The contact-dependent type VI secretion system (T6SS) in diverse microbes plays crucial roles in both inter-bacterial and bacteria-host interactions. As numerous microorganisms inhabit the intestinal ecosystem at a high density, it is necessary to consider the functions of T6SS in intestinal bacteria. In this mini-review, we discuss T6SS-dependent functions in intestinal microbes, including commensal microbes and enteric pathogens, and list experimentally verified species of intestinal bacteria containing T6SS clusters. Several seminal studies have shown that T6SS plays crucial antibacterial roles in colonization resistance, niche occupancy, activation of host innate immune responses, and modulation of host intestinal mechanics. Some potential roles of T6SS in the intestinal ecosystem, such as targeting of single cell eukaryotic competitors, competition for micronutrients, and stress resistance are also discussed. Considering the distinct activities of T6SS in diverse bacteria residing in the intestine, we suggest that T6SS research in intestinal microbes may be beneficial for the future development of new medicines and clinical treatments. [ABSTRACT FROM AUTHOR]

The isolation of stool pathogens is difficult due to their fastidious nature and the rapid overgrowth of fecal flora. In this study, we evaluate the preservation and isolation of enteric pathogens from stool using the automated COPAN Walk-Away Specimen Processor (WASP®) in conjunction with FecalSwab™ and selenite media. Pathogen viability and fecal commensal abundance were stable in FecalSwab™ media under both room-temperature and refrigerated incubation conditions, resulting in a significantly increased number of well-isolated pathogen colonies observed when compared to samples incubated in modified Cary–Blair media. Isolation of individual pathogen colonies was improved via WASP® planting compared to those planted using the Isoplater system. Furthermore, preincubation using the newly formulated COPAN selenite media significantly enhanced the yield of Salmonella enterica serovar Typhimurium. Together, the automated WASP® system combined with FecalSwab™ and selenite media represents a rapid and efficient approach for the processing of stool specimens compared to standard methods. [ABSTRACT FROM AUTHOR]

During intestinal inflammation, host nutritional immunity starves microbes of essential micronutrients, such as iron. Pathogens scavenge iron using siderophores, including enterobactin; however, this strategy is counteracted by host protein lipocalin-2, which sequesters iron-laden enterobactin. Although this iron competition occurs in the presence of gut bacteria, the roles of commensals in nutritional immunity involving iron remain unexplored. Here, we report that the gut commensal Bacteroides thetaiotaomicron acquires iron and sustains its resilience in the inflamed gut by utilizing siderophores produced by other bacteria, including Salmonella , via a secreted siderophore-binding lipoprotein XusB. Notably, XusB-bound enterobactin is less accessible to host sequestration by lipocalin-2 but can be "re-acquired" by Salmonella , allowing the pathogen to evade nutritional immunity. Because the host and pathogen have been the focus of studies of nutritional immunity, this work adds commensal iron metabolism as a previously unrecognized mechanism modulating the host-pathogen interactions and nutritional immunity. [Display omitted] • Commensal iron acquisition impacts host-pathogen competition for iron • Bacteroides thetaiotaomicron acquires iron by siderophore-binding lipoprotein XusB • XusB shields enterobactin from host sequestration • Salmonella exploits XusB-bound enterobactin to evade nutritional immunity Gut commensal B. thetaiotaomicron acquires iron in the inflamed gut via the secreted siderophore-binding lipoprotein XusB. XusB-bound enterobactin is shielded from host sequestration by lipocalin-2 but can be "re-captured" by Salmonella. Spiga et al. implicate commensal iron acquisition as a previously unrecognized layer of nutritional immunity. [ABSTRACT FROM AUTHOR]