Search Results (943)

Background: Antimicrobial resistance (AMR) is a growing global health threat, with children in low- and middle-income countries bearing a disproportionate burden. Data on resistance patterns and diagnostic practices in pediatric populations remain limited. This study evaluated diagnostic utilization and AMR among children hospitalized with bacterial infections at a national referral hospital in Kenya. Methods: We conducted a retrospective cohort study of pediatric inpatients (0–12 years) admitted with bacterial infections between 2017 and 2021. Patient records were identified using ICD-10 codes and reviewed for diagnostic testing and antimicrobial susceptibility. Descriptive statistics were conducted to show infection counts, diagnostic testing, and resistance outcomes. Results: Among 1608 patients, 1009/1608 (63%) were infants under one year. Culture was conducted in 640/1608 (40%) and antimicrobial sensitivity testing in 111/640 (17%) patients. Gastroenteritis (46%) was the most common infection and blood the most frequently collected specimen (31%). Of 1039 cultured specimens, 896/1039 (86%) showed no growth. The most commonly isolated organisms were Klebsiella pneumoniae 19/128 (15%), Staphylococcus epidermidis (13%, 17/128), and Enterococcus faecium (13%, 16/128). Notably, K. pneumoniae showed 100% resistance to third-generation cephalosporins, suggestive of ESBL production. Among the tested samples, 92/128 (72%) had MDROs, and 26/92 (28%) were extensively drug-resistant (XDR). Among the patients tested, 84/111 (76%) had MDROs, of which 25/84 (30%) were XDR. Children under 5 years had higher odds (OR = 5.84, 95% CI: 1.17-38.21) of having MDRO infections, as well as those with multiple admissions (OR = 3.77, 95% CI: 1.06–20.34). Further, increasing age was inversely associated with MDRO presence. The odds of MDRO infection decreased by 24% for every year increase in age (aOR = 0.76; 95% CI: 0.60–0.93; p = 0.006). Conclusions: The findings highlight the limited diagnostic use and a high burden of MDROs and XDR infections in hospitalized children. Strengthening diagnostic capacity and pediatric antimicrobial stewardship is urgently needed in such settings. Full article

Bovine Norovirus (BNoV) is a member of the enterovirus family that can cause gastroenteritis in calves. This virus poses a significant risk to calf growth and development as well as to the long-term sustainability of the cattle industry in China and elsewhere. No specific treatment or vaccine is currently available; thus, the development of a safe and effective vaccine is paramount. Here, we describe a strategy to assemble BNoV virus-like particles (VLPs) using the insect baculovirus expression system (BEV) to express the major structural protein, VP1, and demonstrate their potentiality as vaccines. The results showed that the BNoV-VLP self-assembled into complete spherical particles with a diameter of approximately 40 nm. When it was immunized in mice, the levels of specific IgG and IgA antibodies peaked at weeks 6 and 7 post-immunization, respectively, with maximum titers of 1:25,600 and 1:200. Moreover, we observed a significant increase in the CD4⁺/CD8⁺ T-cell ratio in splenic lymphocytes of immunized mice (p < 0.05), accompanied by a significant increase in TNF-α⁺CD4⁺ T-cells and TNF-α⁺CD8⁺ T-cells (p < 0.05). These results demonstrate that BNoV-VLPs are promising vaccine candidates for providing immunoprotection in the future. These studies support the significant practical implications of using a scientific basis for the development of a BNoV-VLP vaccine. Full article

Anthropogenic warming of the world’s oceans is not just an environmental crisis, but may result in a significant threat to human health. The combination of a warming ocean and increased human activity in coastal waters sets the stage for increased pathogenic Vibrio–human interaction. Warming patterns due to climate change have already been related to the emergence of Vibrio outbreaks in temperate and cold regions. Seafoods, including seaweeds, are uniquely poised to contribute to global food and nutrition security. In recent years there has been a resurgence of interest in seaweeds due to their many uses, high nutritional value, and ability to provide ecosystem services such as habitat provision, carbon and nutrient uptake, and coastal protection. However, some seaweed species can be a reservoir for harbouring pathogenic Vibrio, and illnesses like gastroenteritis have recently been associated with foods prepared with seaweeds. In this study, we investigated the impact of elevated water temperatures on abundances of the major human pathogens Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio vulnificus/cholerae on seaweed and in coastal waters. Three seaweed species, Fucus serratus, Palmaria palmata, and Ulva spp., were exposed to temperature treatments (16 °C and 20 °C) to assess the effects of mean-temperature rise on Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio vulnificus/cholerae colonisation. Colony-forming units (CFUs) on seaweed surfaces and in surrounding water were counted. F. serratus and P. palmata showed significantly higher Vibrio abundances at higher temperatures compared with Ulva spp.; however, temperature did not significantly affect abundances of tested Vibrio species in surrounding waters. These results indicate that certain seaweed species may serve as major hotspots for human pathogenic bacteria in warmer conditions, with implications for human health. Full article

In Brazil, molecular surveillance expanded after Rotarix™ vaccine introduction, alongside G2P[4] dominance. The G2P[6] genotype, despite sharing the same DS-1-like constellation as G2P[4] strains, remains rare. This retrospective study analyzed eight Brazilian G2P[6] strains (2012–2014) through RT-PCR and 11-segments sequencing, followed by phylogenetic analysis. Two distinct groups were identified: 2012–2013 strains (six) carried a DS-1-like backbone with the rare NSP4 E6 genotype, while 2014 strains (two) exhibited the classical DS-1-like constellation with E2. Phylogenetic analysis confirmed the two main clusters: 2012–2013 strains related to classical G2P[4] and uncommon global genotypes, and 2014 strains resembling emerging DS-1-like G1/G3/G8P[8] reassortants. The 2012–2013 strains clustered within G2-VP7 Lineage IVa, while the 2014 strains belonged to Lineage V, reflecting the global distribution of these variants. All VP4 genes were classified within the P[6]-Ia lineage, with phylogenetic analyses suggesting separate introductions from Asia and Africa. The E6 NSP4 gene segment identified in these strains has an undetermined origin and was not previously associated with G2P[6] strains in Brazil. Despite similarities to G2P[4], G2P[6] strains remain rare, with no genomic features explaining their limited spread. Phylogenetic data indicate multiple reassortment events and international viral exchange, highlighting Brazil’s role in RVA diversity. Ongoing full-genome surveillance is crucial to track rare variants and assess their public health relevance. Full article

The cross-species spillover of coronaviruses is considered a serious public health risk. Feline coronavirus (FCoV), canine coronavirus (CCoV), and transmissible gastroenteritis virus (TGEV) are all classified under Alphacoronavirus suis and infect companion animals and livestock. Due to their frequent contact with humans, these viruses pose a potential risk of future cross-species transmission. Molnupiravir, a prodrug of N4-hydroxycytidine, exhibits potent antiviral activity against SARS-CoV-2, a member of the Betacoronavirus genus, and has been approved for the treatment of COVID-19. Molnupiravir was recently shown to be effective against FCoV, suggesting broad-spectrum antiviral activity across coronavirus lineages. Based on these findings, the present study investigated whether molnupiravir is also effective against CCoV and TGEV, which belong to the same Alphacoronavirus suis species as FCoV. We examined the in vitro antiviral effects of molnupiravir using four viral strains: FCoV-1 and -2, CCoV-2, and TGEV. Molnupiravir inhibited plaque formation, viral antigen expression, the production of infectious viral particles, and viral RNA replication in a dose-dependent manner in all strains. IC₅₀ values for CCoV-2 and TGEV, calculated using a feline-derived cell line (fcwf-4), were significantly lower than those for FCoV, suggesting higher sensitivity to molnupiravir. These results demonstrate that molnupiravir exhibited broad antiviral activity against animal coronaviruses classified under Alphacoronavirus suis, providing a foundation for antiviral strategies to mitigate the future risk of cross-species transmission. Full article

Objectives: Saccharomyces boulardii CNCM I-745, a probiotic yeast, is effectively used for the treatment of acute diarrhea as well as for the prevention and treatment of traveller‘s diarrhea and diarrhea under tube feeding. The underlying mechanisms are not fully elucidated. Both antitoxic and regulatory effects on the intestinal barrier, mediated either by the yeast or yeast-derived substrates, have been discussed. Methods: To examine the effects of Saccharomyces boulardii released substrates (S.b.S) on gastrointestinal (GI) barrier function, a murine small intestinal organoid cell model under stress was used. Stress was induced by lipopolysaccharide (LPS) exposure or withdrawal of growth factors from cell culture medium (GF_Red). Stressed organoids were treated with S.b.S (200 µg/mL), and markers of GI barrier and inflammatory response were assessed. Results: GF_Red-induced stress was characterized by disturbances in selected tight junction (TJ) (p < 0.05), adherent junction (AJ) (p < 0.001), and mucin (Muc) formation (p < 0.01), measured by gene expressions, whereby additional S.b.S treatment was found to reverse these effects by increasing Muc2 (from 0.22 to 0.97-fold change, p < 0.05), Occludin (Ocln) (from 0.37 to 3.5-fold change, p < 0.0001), and Claudin (Cldn)7 expression (from 0.13 ± 0.066-fold change, p < 0.05) and by decreasing Muc1, Cldn2, Cldn5, and junctional adhesion molecule A (JAM-A) expression (all p < 0.01). Further, S.b.S normalized expression of nucleotide binding oligomerization domain (Nod)2- (from 44.5 to 0.51, p < 0.0001) and matrix metalloproteinase (Mmp)7-dependent activation (from 28.3 to 0.02875 ± 0.0044 ** p < 0.01) of antimicrobial peptide defense and reduced the expression of several inflammatory markers, such as myeloid differentiation primary response 88 (Myd88) (p < 0.01), tumor necrosis factor α (Tnfα) (p < 0.01), interleukin (IL)-6 (p < 0.01), and IL-1β (p < 0.001). Conclusions: Our data provide new insights into the molecular mechanisms by which Saccharomyces boulardii CNCM I-745-derived secretome attenuates inflammatory responses and restores GI barrier function in small intestinal organoids. Full article

Objectives: Norovirus outbreaks occur in densely populated environments, such as long-term care facilities, hospitals, and schools. On 22 October 2022, an outbreak of acute gastroenteritis was reported at a kindergarten in Korea. An epidemiologic investigation was conducted to identify the source of the infection and prevent further spread. Methods: Rectal swab and environmental samples were collected for bacterial and viral testing. A retrospective cohort study was conducted among 114 kindergarteners at the kindergarten. Relative risks (RRs) and 95% confidence intervals (CIs) were calculated to assess associations of contact with the primary case, as well as food and water consumption. Results: Of the kindergarteners, 28 out of 114 (24.6%) met the case definition. The primary case occurred on 19 October, and subsequent cases began on 21 October. Sharing the same four-year-old class as the primary case (RR, 2.56; 95% CI, 1.35–4.87), being in the same regular class (RR, 2.37; 95% CI, 1.27–4.41), being on the same floor during after-school class (RR, 3.49; 95% CI, 1.74–7.00), and attending the same English class (RR, 1.98; 95% CI, 1.05–3.72) were statistically significant. Consumption of drinking water on the third floor and fourth floor on 20 October had significantly higher and lower RRs, respectively. Norovirus was detected in 9 out of 18 rectal swab samples (50.0%). Conclusions: This norovirus outbreak at the kindergarten was presumed to have been caused by person-to-person transmission from the primary case. Isolation and restriction of symptomatic children in kindergartens should be thoroughly implemented. Additionally, enhanced surveillance among family members of affected individuals is necessary to prevent further outbreaks. Full article

Background/Objectives: Norovirus is a major cause of acute gastroenteritis worldwide. Considering its highly infectious and transmissible nature, rapid and accurate diagnostic tools are of utmost importance for the effective control of outbreaks in the context of point-of-care testing (POCT). In this study, we developed a genogroup-specific multiplex reverse transcriptase loop-mediated isothermal amplification assay to detect the human norovirus genogroups I and II (GI and GII, respectively). Methods: For the comprehensive detection of clinically relevant genotypes, two sets of primers were incorporated into the assays targeting the RdRp-VP1 junction: one against GI.1 and GI.3, and the other for GII.2 and GII.4. Following optimization of the reaction variables, we standardized the reaction conditions at 65 °C with 6 mM MgSO₄, 1.4 mM dNTPs, 7.5 U WarmStart RTx Reverse Transcriptase, and Bst DNA polymerase at 8 U and 10 U for GI and GII, respectively. Amplification was monitored in real-time using a thermocycler platform to ensure precise quantification and detection. Finally, the assay was evaluated through portable isothermal detection device to test its feasibility in on-site settings. Results: Both assays detected the template down to 10²–10³ copies per reaction and showed high target selectivity, yielding no non-specific amplification across 39 enteric pathogens. These assays enabled prompt detection of GI within 10–12 min and of GII within 12–17 min after the reaction was initiated. Onsite validation reveals all template detection below 15 min, demonstrating its potential feasibility in point-of-care applications. Including the sample preparation time, test results were obtained in less than 1 h. Conclusions: This method is a rapid, reliable, and scalable solution for detecting human norovirus in POCT settings for both clinical diagnosis and public health surveillance. Full article

Salmonella enterica serovar Typhimurium (S. Typhimurium) is an intracellular pathogen capable of surviving and replicating within macrophages, which causes foodborne diseases such as gastroenteritis. To develop a strategy against intracellular bacteria in macrophages, we designed silver-loaded biodegradable polymer nanoparticles functionalized with S. Typhimurium membrane vesicles (MVs). Silver nanoparticles (Ag NPs) were initially encapsulated within biodegradable poly(lactic-co-glycolic) nanoparticles (Ag-P NPs), which were then surface-modified with polyethyleneimine to form Ag-PP NPs. These were subsequently fused with S. Typhimurium MVs via a sonication method to generate Ag-PP@MV NPs. The resulting MV-coated nanoparticles displayed a similar protein profile to that of native MVs and exhibited antibacterial activity against intracellular S. Typhimurium. Notably, the enhanced cellular uptake of the MV-modified NPs contributed to their intracellular bactericidal efficacy. This study highlights MV modification as a promising strategy to improve NP delivery to macrophages for treating persistent intracellular infections. Full article

Vibrio parahaemolyticus is the leading bacterial cause of gastroenteritis associated with aquatic food consumption globally. This study aimed to determine the prevalence of V. parahaemolyticus in aquatic foods from Huzhou and to identify the serotypes, antimicrobial resistance, virulence factors, and genetic relatedness of the strains. A total of 306 isolates were detected from 1314 aquatic food samples from 2022 to 2024. The results indicated that the most prevalent serotypes were O1:KUT (17.0%), O2:K28 (13.7%), and O2:KUT (13.1%). Multilocus sequence typing analysis divided the 306 isolates into 175 sequence types (STs), and the predominant sequence type was ST864 (3.3%). Antimicrobial susceptibility tests showed that 2.6% of isolates were multidrug resistant. High resistance was observed to ampicillin (64.7%) and streptomycin (44.4%). A total of seven antimicrobial categories of resistance genes were identified, and the resistance gene bla_CARB was detected in all isolates. The virulence genes tdh and trh were found in 16 (5.2%) and 12 (3.9%) isolates, respectively. In addition, we observed that all the 306 V. parahaemolyticus isolates encode type III secretion systems 1. The phylogenomic analysis based on the whole-genome sequence revealed that the 306 isolates were divided into four clusters. Our findings broaden perspectives on V. parahaemolyticus genetic diversity and enhance our ability to assess the potential risks of its spread. Full article

Pancreatic cancer is an aggressive malignancy, with a current 5-year survival rate in the United States of approximately 13.3%. Although the current standard for resectable pancreatic cancer most commonly includes neoadjuvant chemotherapy prior to a curative resection, surgery, in the majority of patients, has historically been palliative. The latter interventions include open or laparoscopic bypass of the bile duct or stomach in cases of obstructive jaundice or gastric outlet obstruction, respectively. Non-surgical interventional therapies started with percutaneous transhepatic biliary drainage (PTBD), both as a palliative maneuver in unresectable patients with obstructive jaundice and to improve liver function in patients whose surgery was delayed. Likewise, interventional radiologic techniques included the placement of plastic and ultimately self-expandable metal stents (SEMSs) through PTBD tracts in patients with unresectable cancer as well as percutaneous cholecystostomy in patients who developed cholecystitis in the context of malignant obstructive jaundice. Endoscopic retrograde cholangiopancreatography (ERCP) and stent placement (plastic/SEMS) were subsequently used both preoperatively and palliatively, and this was followed by, or undertaken in conjunction with, endoscopic gastro-duodenal SEMS placement for gastric outlet obstruction. Although endoscopic ultrasound (EUS) was initially used to cytologically diagnose and stage pancreatic cancer, early palliation included celiac block or ablation for intractable pain. However, it took the development of lumen-apposing metal stents (LAMSs) to facilitate a myriad of palliative procedures: cholecystoduodenal, choledochoduodenal, gastrohepatic, and gastroenteric anastomoses for cholecystitis, obstructive jaundice, and gastric outlet obstruction, respectively. In this review, we outline these procedures, which have variably supplanted surgery for the palliation of pancreatic cancer in this rapidly evolving field. Full article

A key feature that differentiates Gram-positive and Gram-negative bacteria is the outer membrane, an asymmetric membrane composed of lipopolysaccharides, phospholipids, lipoproteins and integral proteins, including the outer-membrane proteins (OMPs). By being in direct contact with the extracellular milieu, the outer membrane and OMPs participate in multiple functions in Gram-negative bacteria, including controlling nutrient and molecule access to the cytoplasm, membrane vesicle formation and resistance to environmental stresses. OMPs have a characteristic barrel shape formed by antiparallel β-strands, with or without channels that allow diffusion of substrates through the outer membrane. The marine bacterium Vibrio cholerae is responsible for non-invasive gastroenteritis and cholera disease by consumption of contaminated water or food. Its OMPs, besides having a porin function, contribute to resistance to osmotic pressure and antimicrobial agents, intracellular signaling, adhesion to host cells and biofilm formation, amongst other functions. In this review, in addition to quickly reviewing the general structure of the outer membrane, the OMPs and how they reach the outer membrane, the functions attributed to these proteins are compiled. The mechanisms used by each of the described OMP to accomplish these functions in the marine pathogenic bacterium V. cholerae are discussed. Potential clinical and bioengineering applications of OMPs, such as diagnostic tools, vaccine development, and targeted antimicrobial or anti-virulence strategies are presented. What is known about the OMPs of V. cholerae is presented below. Full article

This study aimed to investigate the prevalence, antimicrobial resistance (AMR), and virulence characteristics of Vibrio parahaemolyticus (V. parahaemolyticus) isolated from olive flounder (Paralichthys olivaceus) and rockfish (Sebastes schlegelii) sashimi samples sold in South Korea from 2021 to 2022. A total of 500 fish samples were analyzed, from which 17 V. parahaemolyticus isolates were obtained. Antibiotic susceptibility testing using the minimum inhibitory concentration method revealed that 58.8% (10/17) of the isolates exhibited resistance to ampicillin, indicating the potential for AMR transmission in seafood-associated pathogens. Whole-genome sequencing (WGS) and a polymerase chain reaction detected the presence of tlh and trh virulence genes in all isolates, suggesting their pathogenic potential. Although the overall isolation rate of V. parahaemolyticus was low, the presence of virulence and AMR genes indicates public health relevance associated with raw seafood consumption. The increasing consumer demand for raw fish, coupled with environmental changes such as rising ocean temperatures, underscores the necessity of continuous surveillance to prevent foodborne outbreaks. These findings emphasize the need for targeted AMR monitoring and further research to mitigate the dissemination of resistant V. parahaemolyticus strains and enhance seafood safety. Full article

Waterborne illnesses, including those caused by Salmonella, are an increasing public health challenge, particularly in developing countries. Potential sources of salmonellosis include fruits and vegetables irrigated/treated with surface water, leading to human infections. Salmonella causes millions of gastroenteritis cases annually, but early detection through routine water quality surveillance is time-consuming, requires specialized equipment, and faces limitations, such as coverage gaps, delayed data, and poor accessibility. Climate change-driven extreme events such as floods and droughts further exacerbate variability in water quality. In this context, remote sensing offers an efficient and cost-effective alternative for environmental monitoring. This study evaluated the potential of Sentinel-2 satellite imagery to predict Salmonella occurrence in the Maipo and Mapocho river basins (Chile) by integrating spectral, microbiological, climatic, and land use variables. A total of 1851 water samples collected between 2019 and 2023, including 704 positive samples for Salmonella, were used to develop a predictive model. Predicting Salmonella in surface waters using remote sensing is challenging for several reasons. Satellite sensors capture environmental proxies (e.g., vegetation cover, surface moisture, and turbidity) but not pathogens. Our goal was to identify proxies that reliably correlate with Salmonella. Twelve spectral indices (e.g., NDVI, NDWI, and MNDWI) were used as predictors to develop a predictive model for the presence of the pathogen, which achieved 59.2% accuracy. By spatially interpolating the occurrences, it was possible to identify areas with the greatest potential for Salmonella presence. NDWI and AWEI were most strongly correlated with Salmonella presence in high-humidity areas, and spatial interpolation identified the higher-risk zones. These findings reveal the challenges of using remote sensing to identify environmental conditions conducive to the presence of pathogens in surface waters. This study highlights the methodological challenges that must be addressed to make satellite-based surveillance an accessible and effective public health tool. By integrating satellite data with environmental and microbiological analyses, this approach can potentially strengthen low-cost, proactive environmental monitoring for public health decision-making in the context of climate change. Full article

Background: Acute viral gastroenteritis (AVG) still has an impact on children under 5 years old in developing countries. In Romania, vaccination against Rotavirus is not included in the National Immunization Program. Children from poor social backgrounds represent 48% of the patients hospitalized in Children’s Clinical Hospital of Brasov. The use of antibiotics in Romania is high. Methods: The retrospective study enrolled 1054 children, 0–48 months of age, hospitalized in the Emergency Clinical Hospital for Children Brasov between January 2022–December 2023, for Rotavirus, Adenovirus or Norovirus acute gastroenteritis. Children presenting nosocomial infections were excluded. All children that have met the previously mentioned criteria, presenting positive stool samples (immunochromatography method) for Rotavirus, Adenovirus or Norovirus were included in the study. Results: Out of 1054 cases of AVG 782 (74.2%) were due to Rotavirus, 196 (18.5%) to Adenovirus and 76 (7.3%) to Norovirus. A total of 477 (45.3%) patients came from poor social backgrounds and 577 (54.7%) children from good social backgrounds. Rotavirus infection was equally prevalent in both groups (p < 0.35). Children from poor social backgrounds presented an average age of 11 months (SD ± 9; range 14 days–48 months) and a hospitalization average of 5.86 days (SD ± 3.67; range 1–22 days) compared to the others, who presented an average of 21 months (SD ± 15; range 26 days–48 months) and hospitalization average of 5.20 days (SD ± 2.51; range 1–18 days) (p < 0.01; p < 0.01). Severe dehydration presented 267 children from poor settings (56%) and 186 (17.6%) received antibiotics, compared to the other group—224 children (38.8%) with severe dehydration and 216 (20.4%) receiving antibiotics. The most used antibiotic for both groups was Ceftriaxone (53.9% and 57.6% of all AVG). Conclusions: Rotavirus is the leading AVG cause. Children from poor backgrounds were younger, more dehydrated, hospitalized longer. Full article