Comparative evaluation of two molecular multiplex syndromic panels with acute gastroenteritis.

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.

Enhancing enteric pathogen detection: implementation and impact of multiplex PCR for improved diagnosis and surveillance.

BACKGROUND: Syndromic surveillance of acute gastroenteritis plays a significant role in the diagnosis and management of gastrointestinal infections that are responsible for a substantial number of deaths globally, especially in developing countries. In Lebanon, there is a lack of national surveillance for acute gastroenteritis, and limited data exists regarding the prevalence of pathogens causing diarrhea. The one-year study aims to investigate the epidemiology of common gastrointestinal pathogens and compare our findings with causative agents of diarrhea reported by our study collaborative centers. METHODS: A multicenter, cross-sectional study was conducted over a one-year period. A total of 271 samples were obtained from outpatients and inpatients presenting with symptoms of acute gastroenteritis at various healthcare facilities. The samples were then analyzed using Allplex gastrointestinal assay that identifies a panel of enteric pathogens. RESULTS: Overall, enteropathogens were detected in 71% of the enrolled cases, 46% of those were identified in patients as single and 54% as mixed infections. Bacteria were observed in 48%, parasites in 12% and viruses in 11%. Bacterial infections were the most prevalent in all age groups. Enteroaggregative E. coli (26.5%), Enterotoxigenic E. coli (23.2%) and Enteropathogenic E. coli (20.3%) were the most frequently identified followed by Blastocystis hominis (15.5%) and Rotavirus (7.7%). Highest hospitalization rate occurred with rotavirus (63%), Enterotoxigenic E. coli (50%), Blastocystis hominis (45%) and Enteropathogenic E. coli (43%). Enteric pathogens were prevalent during summer, fall and winter seasons. CONCLUSIONS: The adoption of multiplex real-time PCR assays in the diagnosis of gastrointestinal infections has identified gaps and improved the rates of detection for multiple pathogens. Our findings highlight the importance of conducting comprehensive surveillance to monitor enteric infections. The implementation of a syndromic testing panel can therefore provide healthcare professionals with timely and accurate information for more effective treatment and public health interventions.

Gastrointestinal PCR panel results and antibiotic use in acute gastroenteritis cases: How appropriate are we in our usage?

BACKGROUND: We aimed to determine the pathogens detected by the Gastrointestinal (GI) PCR panel in patients with acute gastroenteritis (AGE), the evaluation of antibiotic use in these patients, and the investigation of the role of laboratory parameters in differentiating viral and bacterial etiologies. METHODS: The demographic characteristics, GI PCR panel results, laboratory investigations, antibiotic usage, and appropriateness of antibiotic treatment were investigated in AGE patients. RESULTS: A total of 175 adult patients with AGE and GI PCR panel results were included in the study. The most common pathogens were EPEC (24.6%) and C. difficile (18.3%). Among the 102 patients receiving antibiotic treatment, 34.3% were evaluated as inappropriate antibiotic use. WBC, CRP, procalcitonin, CRP/albumin ratio, and procalcitonin/albumin ratio were found to be significantly higher in cases with bacterial origin. CONCLUSIONS: The utilization of GI PCR panels in AGE patients has revolutionized the field of diagnostics by providing rapid and accurate identification of pathogens. In units without the possibility of GI PCR testing, CRP, procalcitonin, CRP/albumin ratio and procalcitonin/albumin ratio may be useful in the decision of antibiotic treatment.

Nanozyme-Enhanced Probiotic Spores Regulate the Intestinal Microenvironment for Targeted Acute Gastroenteritis Therapy.

Antibiotic therapeutics to combat intestinal pathogen infections often exacerbate microbiota dysbiosis and impair mucosal barrier functions. Probiotics are promising strategies, because they inhibit pathogen colonization and improve intestinal microbiota imbalance. Nevertheless, their limited targeting ability and susceptibility to oxidative stress have hindered their therapeutic potential. To tackle these challenges, Ces3 is synthesized by in situ growth of CeO2 nanozymes with positive charges on probiotic spores, facilitating electrostatic interactions with negatively charged pathogens and possessing a high reactive oxygen species (ROS) scavenging activity. Importantly, Ces3 can resist the harsh environment of the gastrointestinal tract. In mice with S. Typhimurium-infected acute gastroenteritis, Ces3 shows potent anti-S. Typhimurium activity, thereby alleviating the dissemination of S. Typhimurium into other organs. Additionally, owing to its O2 deprivation capacity, Ces3 promotes the proliferation of anaerobic probiotics, reshaping a healthy intestinal microbiota. This work demonstrates the promise of combining antibacterial, anti-inflammatory, and O2 content regulation properties for acute gastroenteritis therapy.

Comparative evaluation of the detection rate, workflow and associated costs of a multiplex PCR panel versus conventional methods in diagnosis of infectious gastroenteritis.

Introduction. Infectious gastroenteritis is a common reason for consulting a physician. Although most cases of gastrointestinal illness are self-limiting, the identification of the etiologic pathogen by stool specimen analysis is important in cases of more severe illness and for epidemiological reasons.Due to the broad range of causative pathogens, the conventional examination of a stool specimen is labour-intensive and usually requires different diagnostic methods. Multiplex PCR tests [e.g. BioFire Gastrointestinal (GI) Panel] allow the rapid detecting of up to 22 pathogens in one test.Hypothesis. Using a multiplex PCR panel to test stool specimens for infectious gastroenteritis pathogens can improve the detection rate, reduce the time-to-result and hands-on time and lower the costs of a microbiology laboratory.Aim. This study was aimed at evaluating the detection rate, the workflow and associated costs of stool specimen management using the BioFire GI Panel versus conventional methods.Methodology. Stool specimens were evaluated prospectively during the routine operation. Pathogen detection rate, hands-on time, time-to-result and material and personnel costs were determined for the BioFire GI Panel and conventional methods-the latter based on physician request and excluding viral testing.Results. Analysing 333 specimens collected between 2019 and 2020, the detection rate of enteropathogens was significantly higher with a positivity rate of 39.9 % using the multiplex PCR panel compared with 15.0 % using the conventional methods. The BioFire GI Panel presented results in a median time of 2.2 h compared with 77.5 h for culture and 22.1 h for antigen testing, noting that no tests were performed at weekends except for toxinogenic Clostridioides difficile. Based on list prices, the BioFire GI Panel was nine times more expensive compared with conventional methods, whereas hands-on-time was significantly lower using the BioFire GI Panel.Conclusion. Multiplex PCR panels are valuable tools for laboratory identification of infectious agents causing diarrhoea. The higher costs of such a multiplex PCR panel might be outweighed by the higher detection rate, ease of handling, rapid results and most likely improved patient management. However, these panels do not provide information on antimicrobial susceptibility testing. Therefore, if this is necessary for targeted therapy or if outbreak monitoring and control is required, specimens must still be cultured.

Detection and Characterization of Bacterial and Viral Acute Gastroenteritis among Outpatient Children under 5 Years Old in Guangzhou, China.

Acute gastroenteritis (AGE) in children can be attributed to a multitude of bacterial and viral pathogens. The objective of this study was to investigate the epidemiology of bacterial and viral AGE in children and to compare clinical characteristics between single and multiple enteric pathogen infections. A total of 456 stool samples were collected from outpatient children under 5 years old with AGE, which were subsequently analyzed for nine bacteria and three viruses using the Luminex xTAG® Gastrointestinal Pathogen Panel. The presence of at least one pathogen was detected in 260 cases (57.0%), with Salmonella being the predominant agent, followed by norovirus, Campylobacter, and rotavirus. A total of 69 cases (15.1%) exhibited positive results for two or more enteric pathogens. Although certain co-infections demonstrated significant differences in primary clinical features compared with mono-infections, no statistical variance was observed in terms of disease severity. In outpatient children from southern China, Salmonella emerged as the most prevalent causative agent of AGE, succeeded by norovirus and Campylobacter. This study underscores the burden posed by coinfections and highlights the clinical characteristics associated with AGE when accompanied by coinfections among children under 5 years old.

Factors Associated With Actionable Gastrointestinal Panel Results in Hospitalized Children.

OBJECTIVES: There is uncertainty regarding which hospitalized patients with acute gastroenteritis (AGE) benefit from gastrointestinal panel (GIP) testing. Unnecessary testing may lead to increased costs, overdiagnosis, and overtreatment. In general, AGE management and outcomes are most impacted if an actionable (bacterial or parasitic) result is obtained. We aimed to assess which clinical reasons for ordering GIP testing ("order indications") and patient factors were associated with actionable results. METHODS: This is a cross-sectional study of pediatric patients hospitalized between 2015 and 2018 at a large pediatric health care system with diarrhea and a GIP performed. Multivariable regression analysis was used to determine associations between actionable GIP results and order indication, stool frequency, and demographics. Findings were evaluated in patients with complex chronic conditions (CCC) and non-CCC patients. RESULTS: There were 1124 GIPs performed in 967 encounters. Non-CCC patients had more actionable results than CCC patients, and reasons for testing differed. Across both cohorts, age ≥1 year old was positively associated with actionable results. For non-CCC patients, actionable results were associated with "diarrhea with blood or pus" order indication and nonwinter season; international travel was associated with non-Clostridioides difficile bacteria and parasites. No order indications were associated with actionable results for CCC patients. CONCLUSIONS: Patient factors and order indications that may help identify children hospitalized for AGE with actionable GIP results include older age (regardless of CCC status), as well as bloody stools and international travel in previously healthy children. Prospective validation of these findings could help improve diagnostic stewardship and decrease unnecessary testing.

Diagnosis of gastrointestinal infections: comparison between traditional microbiology and a commercial syndromic molecular-based panel.

Traditional diagnosis of infectious gastroenteritis is based on culture, microscopy and antigen detection. The development of gastrointestinal syndromic panels based on molecular techniques have allowed rapid and simultaneous identification of multiple pathogens. The objective was to evaluate the implementation of Allplex™ Gastrointestinal Panel Assays (AGPA): Allplex™ GI-Virus, Allplex™ GI-Bacteria (I) and Allplex™ GI-Parasite by comparing with traditional diagnosis. A retrospective comparative study was conducted at Hospital Universitario La Paz, between the first year of implementation of the AGPA (April 1, 2018 to March 31, 2019) and the results obtained during the previous year with traditional methods (April 1, 2017 to March 31, 2018). With the implementation of AGPA we obtained an increase in the detection of rotavirus and adenovirus, being statistically significant for rotavirus ([CI95%:3.60-6.79]; P < 0.05) and an increase in the positivity rates of all the bacteria tested, with the exception of Salmonella spp. ([CI95%:3.60-6.79]; P < 0.05). Comparing the bacteria recovered by culture, we obtained an increase in the case of Shigella spp. cultivation during the AGPA period. Regarding protozoa, we achieved a significant increase in the positivity rates for Cryptosporidium spp. ([CI95%:1.98-3.01] P < 0.05), Giardia intestinalis ([CI95%:3.94-5.25]; P < 0.05) and Blastocystis spp. ([CI95%:9.44-11.36]; P < 0.05). There was an improvement in report turnaround time when comparing molecular diagnosis to bacterial culture and concentration plus microscopy for parasites; but not compared with antigen detection. The molecular diagnosis approach with AGPA were more sensitive and had a faster turnaround time for some targets, and in our setting, enabled an increased diagnostic capacity for viruses and protozoa.

Campylobacter jejuni and Campylobacter coli in human stool samples: antibiotic resistance profiles, putative virulence determinants and molecular characterization of the isolates.

Campylobacters, especially C. jejuni and C. coli, have become one of the leading causes of acute gastroenteritis in humans worldwide in recent years. We aimed to investigate the presence, antimicrobial resistance, putative virulence genes, and molecular characterization of C. jejuni and C. coli recovered from human acute gastroenteritis cases in the study. In the study, suspected Campylobacter spp. isolates were obtained in 43 (5%) feces samples collected from a total of 850 patients who applied to the Erciyes University Medical Faculty acute clinic between March 2019 and February 2020. As a result of the phenotypic tests, these isolates were determined to be Campylobacter spp. According to the multiplex PCR, 33 of 43 Campylobacter spp. isolates were identified as C. jejuni (76%) and ten isolates were as C. coli (24%). In the disc diffusion test, the highest resistance rate was found in the trimethoprim/sulfamethoxazole (90.1%) and ciprofloxacin (90.1%), and the lowest rate was found in the amoxicillin-clavulanic acid (9.3%). In Campylobacter spp. isolates, the virulence genes cdtA, virB11, cdtB, cadF, iam, ceu, and flaA were found to be positive at rates of 26 (60%), 28 (65.6%), 13 (30%), 4 (9%), 27 (62%), 17 (39%), and 7 (16%), respectively. However, the cdtC gene was not detected in any of the isolates. The study searched tetO gene to examine the genetic aspect of tetracycline resistance, which was found in all Campylobacter spp. isolates. In the PCR reactions to investigate A2074C and A2075G mutations of macrolide resistance, it was determined as 7 (16%) and 21 (48%) of the isolates. To detect quinolone resistance, the rates of quinolone resistance-determining regions (QRDR) were 20 (45.4%) and the gyrA gene mutations in the mismatch amplification mutation assay PCR (MAMA-PCR), were 19 (43.1%) of isolates. In addition, the quinolone resistance gene (qnr) carried by plasmid in Campylobacter isolates was not found in the study. BlaOXA-61 and CmeB (multi-drug efflux pump) genes were detected as 28 (63.6%) and 30 (68.1), respectively. The Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR) used for typing the isolates revealed that the band profiles obtained from the isolates were different. In conclusion, this was a very comprehensive study revealing the presence of antibiotic-resistant C. jejuni and C. coli with various virulence genes in patients admitted to a university hospital with acute gastroenteritis. This is of utmost significance for public health. Since campylobacteria are foodborne, zoonotic pathogens and transmission occurs mostly through food. People should have detailed information about the transmission routes of campylobacteria and risky foods. In addition, staff, food processors and caterers, should be trained in hygiene.

Prevalence and risk factors associated with the occurrence of Campylobacter sp. in children aged 6-24 months in peri-urban Nairobi, Kenya.

Introduction: Campylobacter bacteria is a major cause of foodborne-related bacterial gastroenteritis in humans worldwide. It is known to cause diarrhea in young children which has been shown to directly affect their weight and height as a result of malnutrition. Severe cases of diarrhea can also lead to death. Most of the burden is experienced in resource-limited countries in Africa and Southeast Asia where the disease is linked to poor hygiene and sanitation. The objective of this study was to determine the prevalence of Campylobacter in children aged between 6 and 24 months in Nairobi, Kenya and identify potential risk factors associated with their occurrence. Methods: A cross-sectional study was carried out between May to December 2021. A total of 585 randomly selected households were visited in two wards (Uthiru/Ruthimitu and Riruta) in Dagoretti South sub-county, Nairobi. A questionnaire regarding how children's food is handled, the major foods consumed, sanitation and hygiene, and animal ownership was conducted among caregivers to identify associated risk factors. Stool samples were collected from 540/585 children and screened for the presence of Campylobacter using culture-based methods and confirmed through PCR. Results: Of the 540 children's stool samples processed, Campylobacter isolates were detected in 4.8% (26/540). Campylobacter jejuni (C. jejuni) was the most common species in 80.8% of positive samples compared to Campylobacter coli (C. coli) in 26.9% of samples. In six samples, both C. jejuni and C. coli were isolated, while in four samples, it was not possible to speciate the Campylobacter. Drinking cow's milk (OR 4.2, 95% CI 1.4 - 12.6) and the presence of animal feces in the compound (OR 3.4, 95% CI 1.1 - 10.3) were found to be statistically associated with Campylobacter carriage in children. Discussion: The carriage of Campylobacter in children in this community indicates a need for further investigation on source attribution to understand transmission dynamics and inform where to target interventions. Awareness creation among caregivers on good personal and food hygiene is needed, including boiling milk before consumption. Implementation of biosecurity measures at the household level is highly recommended to reduce contact between animals and humans.

Érase una vez una niña y su lindo gatito / Once upon a time there was a girl and her cute kitty

Introducción: Campylobacter es el principal patógeno de gastroenteritis transmitida por alimentos, ocurriendo generalmente por la ingesta de pollo mal cocinado, constituyendo otra importante fuente de infección los cachorros de animales domésticos. Caso clínico: escolar con gastroenteritis aguda con sospecha diagnóstica inicial de giardiasis por ambiente epidémico (gato doméstico). Se recoge coprocultivo en el que se detecta Campylobacter jejuni, prescribiéndose azitromicina, dado lo prolongado de la clínica. A lo largo del control evolutivo en el centro de salud la familia informa de que se ha solicitado nueva muestra de heces en el gato, dado persistencia de los síntomas pese a tratamiento con metronidazol. Finalmente, crece también Campylobacter jejuni en el coprocultivo de la mascota. Tras finalizar ambos el tratamiento antibiótico, permanecen asintomáticos. Como posible alimento sospechoso del origen del cuadro está el corazón de pollo no cocinado con el que alimentaban al gato de forma habitual. Conclusiones: ante un cuadro de gastroenteritis aguda es fundamental una adecuada anamnesis que incluya ambiente epidémico y alimentos sospechosos. En ocasiones las mascotas también constituyen una fuente de transmisión de la infección a nuestros pacientes. En este caso se sospecha la cadena de contaminación: corazón de pollo no cocinado-heces de gato doméstico-niña (AU)

Introduction: Campylobacter is a well-known food-borne pathogen that causes human gastroenteritis. The most common way for children to become infected with campylobacteriosis is through chicken that is not fully cooked, another important source of infection are domestic puppies.Case report: it is presented the case of an eight-year-old girl with acute gastroenteritis, the first diagnostic suspicion was giardiasis due to epidemic environment (domestic cat). A stool culture was collected in which Campylobacter jejuni was detected. Azithromycin was prescribed because of prolonged symptoms. Throughout the control in the health center, family reported that a new fecal sample has been requested from the cat due to the persistence of the symptoms despite treatment with metronidazole. Finally, Campylobacter jejuni also grew in the pet's stool culture. After both finished antibiotic treatment, they remained asymptomatic. The possible suspected infection source was the chicken heart with which the cat was regularly fed. Conclusions: the evaluation of the child with acute gastroenteritis begins with a careful history which includes epidemiological environment and suspicious food intake. Ocassionally, pets are also a source of transmission to our patients. In this case, the suspected contamination chain was: uncooked chicken heart- domestic cat faeces-girl. (AU)

Anti-infective treatment of gastro-intestinal tract infections in children.

Gastroenteritis is most often viral in origin and Rotavirus and Norovirus most frequently implicated in young children. Stool-based multiplex Polymerase Chain Reaction (PCR) can detect bacteria, viruses or parasites that may or may not be responsible for gastroenteritis (colonization). While the etiological profile of these digestive infections has greatly benefited from PCR, in the absence of underlying pathologies the presence of potential pathogens does not justify anti-infectious treatment. Indeed, very few bacterial causes require antibiotic treatment, apart from shigellosis, severe forms of salmonellosis and a few Campylobacter sp. infections. The development of antibiotic resistance in Salmonella sp., Shigella sp. and Campylobacter sp. is a cause for concern worldwide, limiting therapeutic options. The antibiotics proposed in this guide are in line with the joint recommendations of the European Society of Pediatric Infectious Diseases and the European Society of Pediatric Gastroenterology and Nutrition. Azithromycin is preferentially used to treat infections with Shigella sp. or Campylobacter sp. Ceftriaxone and ciprofloxacin are recommended for salmonellosis requiring antibiotic therapy. Empirical treatments without bacterial identification are not indicated except in cases of severe sepsis or in subjects at risk (e.g., sickle-cell disease). Metronidazole should be prescribed only for acute intestinal amebiasis after microbiological confirmation.

An Unlikely Guest With an Overstayed Welcome: Cyclospora-Induced Postinfectious Irritable Bowel Syndrome.

INTRODUCTION: Postinfectious irritable bowel syndrome is a phenomenon that can occur following bouts of acute gastroenteritis. While bacterial pathogens are typically implicated in the development of postinfectious irritable bowel syndrome, viral and parasitic infections should also be considered as inciting pathogens. CASE PRESENTATION: An immunocompetent, 65-year-old woman presented with several weeks of watery diarrhea, which polymerase chain reaction testing confirmed to be a Cyclospora infection. Resolution of diarrhea was achieved with antibiotic treatment, however, months later she presented to the gastroenterology service with persistence of loose stools and abdominal cramping consistent with a diagnosis of postinfectious irritable bowel syndrome. DISCUSSION: Postinfectious irritable bowel syndrome has a similar presentation to sporadic irritable bowel syndrome, with diagnosis aided by the identification of an inciting pathogen. To our knowledge, this is the first documented case of Cyclospora-induced postinfectious irritable bowel syndrome. While parasitic infections typically aren't implicated in cases of postinfectious irritable bowel syndrome, this case highlights the value of considering this condition as a cause of protracted diarrhea in patients previously diagnosed with Cyclospora.

Enteric Aeromonas Infection: a Common Enteric Bacterial Infection with a Novel Infection Pattern Detected in an Australian Population with Gastroenteritis.

Aeromonas species are emerging human enteric pathogens. However, they are currently not routinely detected in many diagnostic laboratories, and information regarding Aeromonas enteric infections detected using molecular methods is lacking. Here, we investigated the detection of Aeromonas species and four other enteric bacterial pathogens in 341,330 fecal samples from patients with gastroenteritis processed in a large Australian diagnostic laboratory between 2015 and 2019. These enteric pathogens were detected using quantitative real-time PCR (qPCR) methods. Furthermore, we compared the qPCR cycle threshold (CT) values obtained from fecal samples that tested positive for Aeromonas only by molecular detection with those of samples that tested positive by both molecular detection and bacterial isolation methods. Aeromonas species were found to be the second most common bacterial enteric pathogens among patients with gastroenteritis. We observed a unique pattern of three infection peaks for Aeromonas, which correlated with the age of the patients. Aeromonas species were the most common enteric bacterial pathogens in children younger than 18 months. Fecal samples that tested positive for Aeromonas only by molecular detection had significantly higher CT values than fecal samples that tested positive by both molecular detection and bacterial culture. In conclusion, our findings reveal that Aeromonas enteric pathogens exhibit an age-related three-peak infection pattern, distinguishing them from other enteric bacterial pathogens. Moreover, the high rate of Aeromonas enteric infection discovered in this study suggests that Aeromonas species should be routinely tested in diagnostic laboratories. Our data also show that combining qPCR with bacterial culture can enhance the detection of enteric pathogens. IMPORTANCE Aeromonas species are emerging human enteric pathogens. However, these species are currently not routinely detected in many diagnostic laboratories, and no studies have reported the detection of Aeromonas enteric infection using molecular methods. We investigated the presence of Aeromonas species and four other enteric bacterial pathogens in 341,330 fecal samples from patients with gastroenteritis using quantitative real-time PCR (qPCR) methods. Interestingly, we discovered that Aeromonas species were the second most common bacterial enteric pathogens in patients with gastroenteritis, exhibiting a novel infection pattern compared to those of other enteric pathogens. Furthermore, we found that Aeromonas species were the most prevalent enteric bacterial pathogens in children aged 6 to 18 months. Our data also revealed that qPCR methods exhibit higher sensitivity in detecting enteric pathogens compared to that of bacterial culture alone. Moreover, combining qPCR with bacterial culture enhances the detection of enteric pathogens. These findings emphasize the importance of Aeromonas species in public health.

Species Delineation and Comparative Genomics within the Campylobacter ureolyticus Complex.

Campylobacter ureolyticus is an emerging pathogen increasingly appreciated as a common cause of gastroenteritis and extra-intestinal infections in humans. Outside the setting of gastroenteritis, little work has been done to describe the genomic content and relatedness of the species, especially regarding clinical isolates. We reviewed the epidemiology of clinical C. ureolyticus cultured by our institution over the past 10 years. Fifty-one unique C. ureolyticus isolates were identified between January 2010 and August 2022, mostly originating from abscesses and blood cultures. To clarify the taxonomic relationships between isolates and to attribute specific genes with different clinical manifestations, we sequenced 19 available isolates from a variety of clinical specimen types and conducted a pangenomic analysis with publicly available C. ureolyticus genomes. Digital DNA:DNA hybridization suggested that these C. ureolyticus comprised a species complex of 10 species clusters (SCs) and several subspecies clusters. Although some orthologous genes or gene functions were enriched in isolates found in different SCs and clinical specimens, no association was significant. Nearly a third of the isolates possessed antimicrobial resistance genes, including the ermA resistance gene, potentially conferring resistance to macrolides, the treatment of choice for severe human campylobacteriosis. This work effectively doubles the number of publicly available C. ureolyticus genomes, provides further clarification of taxonomic relationships within this bacterial complex, and identifies target SCs for future analysis.

Comparison of a Rapid Multiplex Gastrointestinal Panel with Standard Laboratory Testing in the Management of Children with Hematochezia in a Pediatric Emergency Department: Randomized Controlled Trial.

Advances in diagnostic microbiology allow for the rapid identification of a broad range of enteropathogens; such knowledge can inform care and reduce testing. We conducted a randomized, unblinded trial in a tertiary-care pediatric emergency department. Participants had stool (and rectal swabs if stool was not immediately available) tested using routine microbiologic approaches or by use of a device (BioFire FilmArray gastrointestinal panel), which identifies 22 pathogens with a 1-h instrument turnaround time. Participants were 6 months to <18.0 years and had acute bloody diarrhea. Primary outcome was performance of blood tests within 72 h. From 15 June 2018 through 7 May 2022, 60 children were randomized. Patients in the BioFire FilmArray arm had a reduced time to test result (median 3.0 h with interquartile range [IQR] of 3.0 to 4.0 h, versus 42.0 h (IQR 23.5 to 47.3 h); difference of -38.0 h, 95% confidence interval [CI] of -41.0 to -22.0 h). Sixty-five percent (20/31) of participants in the BioFire FilmArray group had a pathogen detected-most frequently enteropathogenic Escherichia coli (19%), Campylobacter (16%), and Salmonella (13%). Blood tests were performed in 52% of children in the BioFire FilmArray group and 62% in the standard-of-care group (difference of -10.5%, 95% CI of -35.4% to 14.5%). There were no between-group differences in the proportions of children administered intravenous fluids, antibiotics, hospitalized, or who had diagnostic imaging performed. Testing with the BioFire FilmArray reduced the time to result availability by 38 h. Although statistical significance was limited by study power, BioFire FilmArray use was not associated with clinically meaningful reductions in health care utilization or improved outcomes. IMPORTANCE Advances in diagnostic microbiology now allow for the faster and more accurate detection of an increasing number of pathogens. We determined, however, that in children with acute bloody diarrhea, these advances did not necessarily translate into improved clinical outcomes. While a greater number of pathogens was identified using a rapid turnaround multiplex stool diagnostic panel, with a reduction in the time to stool test result of over 1.5 days, this did not alter the practice of pediatric emergency medicine physicians, who continued to perform blood tests on a large proportion of children. While our conclusions may be limited by the relatively small sample size, targeted approaches that educate clinicians on the implementation of such technology into clinical care will be needed to optimize usage and maximize benefits.

Uptake of Tropheryma whipplei by Intestinal Epithelia.

BACKGROUND: Tropheryma whipplei (TW) can cause different pathologies, e.g., Whipple's disease and transient gastroenteritis. The mechanism by which the bacteria pass the intestinal epithelial barrier, and the mechanism of TW-induced gastroenteritis are currently unknown. METHODS: Using ex vivo disease models comprising human duodenal mucosa exposed to TW in Ussing chambers, various intestinal epithelial cell (IEC) cultures exposed to TW and a macrophage/IEC coculture model served to characterize endocytic uptake mechanisms and barrier function. RESULTS: TW exposed ex vivo to human small intestinal mucosae is capable of autonomously entering IECs, thereby invading the mucosa. Using dominant-negative mutants, TW uptake was shown to be dynamin- and caveolin-dependent but independent of clathrin-mediated endocytosis. Complementary inhibitor experiments suggested a role for the activation of the Ras/Rac1 pathway and actin polymerization. TW-invaded IECs underwent apoptosis, thereby causing an epithelial barrier defect, and were subsequently subject to phagocytosis by macrophages. CONCLUSIONS: TW enters epithelia via an actin-, dynamin-, caveolin-, and Ras-Rac1-dependent endocytosis mechanism and consecutively causes IEC apoptosis primarily in IECs invaded by multiple TW bacteria. This results in a barrier leak. Moreover, we propose that TW-packed IECs can be subject to phagocytic uptake by macrophages, thereby opening a potential entry point of TW into intestinal macrophages.

Defining a clinical prediction rule to diagnose bacterial gastroenteritis requiring empirical antibiotics in an emergency department setting: A retrospective review.

BACKGROUND: Gastroenteritis (GE) is a non-specific term for various pathologic states of the gastrointestinal tract. Infectious agents usually cause acute GE. At present, there are no robust decision-making rules that predict bacterial GE and dictate when to start antibiotics for patients suffering from acute GE to the emergency department (ED). We aim to define a clinical prediction rule to aid in the diagnosis of bacterial GE, requiring empirical antibiotics in adult patients presenting to the emergency department with acute GE. METHODS: A two-year retrospective case review was performed on all cases from July 2015 to June 2017 that included patients with acute GE symptoms referred to the ED, after which their stool cultures were performed. The clinical parameters analyzed included patient with comorbid conditions, physical examination findings, historical markers, point-of-care and radiographic tests and other laboratory work. We then used multi-variate logistic regression analysis on each group (bacterial culture-positive GE and bacterial culture-negative GE) to elucidate clinical criteria with the highest yield for predicting bacterial gastroenteritis (BGE). RESULTS: A total of 756 patients with a mean age of 52 years, 52% female and 48% male, respectively, were included in the study. On the basis of the data of these patients, we suggested using a scoring system to delineate the need for empirical antibiotics in patients with suspected bacterial GE based on six clinical and laboratory variables. We termed this the BGE score. A score 0 - 2 points suggests low risk (0.9%) of bacterial GE. A score of 3 - 4 points confers an intermediate risk of 12.0% and a score of 5 - 8 points confers a high risk of 85.7%. A cut-off of  ≥ 5 points may be used to predict culture-positive BGE with a 75% sensitivity and 75% specificity. The area under the receiver operating characteristic (AUROC) for the scoring system (range 0 - 8) was 0.812 (95% CI: 0.780-0.843) p-value < 0.001. CONCLUSION: We suggest using the BGE scoring system (cut-off ≥ 5 points) to delineate the need for empirical antibiotics in patients diagnosed with gastroenteritis. While this is a pilot study, which will require further validation with a larger sample size, our proposed decision-making rule will potentially serve to improve the diagnosis of BGE and thus reduce unnecessary prescription of antibiotics, which will in turn reduce antibiotic-associated adverse events and save on costs worldwide.

A Point Source Gastroenteritis Outbreak in a High School Putatively Due to Clostridium perfringens: Timely Investigation Is Everything.

On May 28, 2021, several gastroenteritis cases were reported among students and staff of a high school on a Greek island. Lunches were delivered daily at the school by a catering company. We conducted a retrospective cohort study. Cases were defined as students or school staff presenting with diarrhea and abdominal pain between May 27 and 29, 2021. We used a self-administered questionnaire to collect the data. In the univariate analysis, we calculated food-specific attack rates (ARs), relative risks (RRs), and confidence intervals (95% CIs). Statistically significant variables (p < 0.05) were included in the multivariable model. Stool samples were collected from students on the same day of notification and were tested for bacteria, viruses, and parasites. The local food safety authority also inspected the school and the catering company premises. The food samples were sent to the reference laboratory for analysis. We received 129 responses from 174 students and staff members (response rate: 74%). Thirty cases were identified (AR, 23%). Of the 11 significant food items in the univariate analysis, the consumption of spaghetti with minced meat on May 27 (odds ratio 53.40, 95% CI: 3.9-736.1, 97% of cases exposed) was independently associated with the onset of gastroenteritis. Clostridium perfringens was detected in two of the three students' stool samples. Six samples from spaghetti with minced meat were positive for C. perfringens and four of those also tested positive for Bacillus cereus. Environmental investigation revealed non-compliances concerning food personnel training and Hazard Analysis and Critical Control Points (HACCP) plan implementation in the food establishment. Timely investigation and samples collection allowed the identification of the probable source of the outbreak. Changes, regarding the reception and distribution of meals in the school, were applied. Recommendations were given to the catering establishment focusing on adequate personnel training and HACCP issues, especially regarding temperature/time controls during all production, handling, and distribution phases.

Bacterial Infections and Cancer: Exploring This Association And Its Implications for Cancer Patients.

Bacterial infections are common in the etiology of human diseases owing to the ubiquity of bacteria. Such infections promote the development of periodontal disease, bacterial pneumonia, typhoid, acute gastroenteritis, and diarrhea in susceptible hosts. These diseases may be resolved using antibiotics/antimicrobial therapy in some hosts. However, other hosts may be unable to eliminate the bacteria, allowing them to persist for long durations and significantly increasing the carrier's risk of developing cancer over time. Indeed, infectious pathogens are modifiable cancer risk factors, and through this comprehensive review, we highlight the complex relationship between bacterial infections and the development of several cancer types. For this review, searches were performed on the PubMed, Embase, and Web of Science databases encompassing the entirety of 2022. Based on our investigation, we found several critical associations, of which some are causative: Porphyromonas gingivalis and Fusobacterium nucleatum are associated with periodontal disease, Salmonella spp., Clostridium perfringens, Escherichia coli, Campylobacter spp., and Shigella are associated with gastroenteritis. Helicobacter pylori infection is implicated in the etiology of gastric cancer, and persistent Chlamydia infections present a risk factor for the development of cervical carcinoma, especially in patients with the human papillomavirus (HPV) coinfection. Salmonella typhi infections are linked with gallbladder cancer, and Chlamydia pneumoniae infection is implicated in lung cancer, etc. This knowledge helps identify the adaptation strategies used by bacteria to evade antibiotic/antimicrobial therapy. The article also sheds light on the role of antibiotics in cancer treatment, the consequences of their use, and strategies for limiting antibiotic resistance. Finally, the dual role of bacteria in cancer development as well as in cancer therapy is briefly discussed, as this is an area that may help to facilitate the development of novel microbe-based therapeutics as a means of securing improved outcomes.