Search Results (293)

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

Zoonotic opportunistic enteric protozoa represent a significant global health threat to immunocompromised populations, especially individuals with human immunodeficiency virus (HIV). Despite China’s severe HIV burden, molecular epidemiological data on enteric protozoa remain limited in this population. In this study, we investigated the occurrence and molecular characteristics of Cryptosporidium species, Giardia duodenalis, and Enterocytozoon bieneusi among 150 AIDS patients with severe immunodeficiency in Wuhan city, Hubei Province, China. The overall test-positive rate was 5.33% (8/150), comprising Cryptosporidium species (including C. hominis, C. parvum, and C. meleagridis) in 2.00% (3/150) and G. duodenalis (including assemblage A, B, and E) in 3.33% (5/150); E. bieneusi was not detected. Notably, this study reports the first identification of the C. parvum subtype IIcA5G3 in humans in China, certainly indicating possible cross-border transmission. Furthermore, the detection of C. meleagridis IIIbA22G1R1c provided additional molecular evidence for chicken-to-human transmission. The finding of G. duodenalis assemblage E highlights the underrecognized zoonotic spillover risks to immunocompromised populations. These findings emphasize the diversity of infectious reservoirs, and the need for enhanced national molecular surveillance of these neglected zoonotic enteric protozoa, alongside targeted interventions for vulnerable populations. Full article

Objective: This proof-of-concept study aimed to evaluate and compare the clinical performance of two AI language models (ChatGPT-3.5 and OpenAI o3) in answering clinical pharmacy multiple-choice questions (MCQs), benchmarked against responses from specialist clinical pharmacists in Jordan, including academic preceptors and hospital-based clinicians. Methods: A total of 60 clinical pharmacy MCQs were developed based on current guidelines across four therapeutic areas: cardiovascular, endocrine, infectious, and respiratory diseases. Each item was reviewed by academic and clinical experts and then pilot-tested with five pharmacists to determine clarity and difficulty. Two ChatGPT models—GPT-3.5 and OpenAI o3—were tested using a standardized prompt for each MCQ, entered in separate sessions to avoid memory retention. Their answers were classified as true/false positives or negatives and retested after two weeks to assess reproducibility. Simultaneously, 25 licensed pharmacists (primarily from one academic institution and several hospitals in Amman) completed the same MCQs using validated references (excluding AI tools). Accuracy, sensitivity, specificity, and Cohen’s Kappa were used to compare AI and human performance, with statistical analysis conducted using appropriate tests at a significance level of p ≤ 0.05. Results: OpenAI o3 achieved the highest accuracy (83.3%), sensitivity (90.0%), and specificity (70.0%), outperforming GPT-3.5 (70.0%, 77.5%, 55.0%) and pharmacists (69.7%, 77.0%, 55.0%). AI performance declined significantly with increasing question difficulty. OpenAI o3 showed the highest accuracy in the cardiovascular domain (93.3%), while GPT-3.5 performed best in infectious diseases (80.0%). Reproducibility was higher for GPT-3.5 (81.6%, κ = 0.556) than OpenAI o3 (76.7%, κ = 0.364). Over two test rounds, GPT-3.5’s accuracy remained stable, whereas OpenAI o3’s accuracy decreased from 83.3% to 70.0%, indicating some variability. Conclusions: OpenAI o3 shows strong promise as a clinical decision-support tool in pharmacy, especially for low- to moderate-difficulty questions. However, inconsistencies in reproducibility and limitations in complex cases highlight the importance of cautious, supervised integration alongside human expertise. Full article

Necrotic enteritis, caused by Clostridium perfringens (C. perfringens) is a disease present worldwide and causes major economic losses. The re-emergence of the disease, in recent years, is mainly due to the ban of the usage of antibiotics as growth promoters in the EU. The aim of this study was to establish a reliable, robust challenge model. Ross hybrid broilers were divided into randomized groups: a positive and a negative control group, a group receiving antibiotic treatment and three groups fed with assorted feed supplements, all receiving the same basal diet. The birds in the treatment groups were vaccinated twice using a 10-times dose of an Infectious Bursitis live vaccine and the animals were challenged four times with a NetB toxin producing C. perfringens strain. The presence of clinical signs and body weight gain were monitored. At the end of the study necropsy was performed and the gut lesions were scored. During the experiment, clinical signs were absent in the negative control group and in the antibiotic treated group. The other animals displayed diarrhea and feather loss. These symptoms were the most pronounced in the positive control group. The gut lesion scores showed significant differences between the negative and positive control groups, with the former scoring the lowest. Based on these results, the challenge model establishment was successful and in this setup the assessment of the potency of feed additives is also possible. Full article

Serious enteric disease caused by seven species of Eimeira continues to cause significant economic damage to the poultry industry. E. acervulina is one of the most widespread strains in farms and has a significant impact on chicken weight loss. Currently, the use of anticoccidial agents to suppress the occurrence of coccidiosis in farms is considerably restricted due to public health and environmental pollution issues. It is important to understand the protective immunity of the host against Eimeria infections with regard to natural products that could be used as alternatives to anticoccidial agents. Berberine chloride is known for its various biological functions, including its anti-parasite activity. However, its impact on intestinal morphology and immune-related activity in broilers infected with Eimeria still remains unclear. The aim of this study is to evaluate the anticoccidial effects of a berberine-based diet in broilers infected with E. acervulina and to monitor the host immune phenomenon using transcriptomic analysis. Administration of berberine to chickens infected with E. acervulina significantly reduced fecal oocyst production and intestinal lesion scores, and increased duodenal villus height, indicating anticoccidial activity and positive effects on intestinal morphology. Transcriptomic analysis of chickens infected with E. acervulina generally observed the down-regulation of metabolism-related genes and the up-regulation of cell integrity-related genes at day 4 post-infection. At day 6 post-infection, an increase in immune-related genes and cellular-homeostasis-related genes was generally observed. Berberine-treated and E. acervulina-infected chickens showed cytokine-cytokine receptor interaction in the second term in a Kyoto Encyclopedia of Genes and Genomes pathway analysis at day 4 post-infection, but not in chickens infected with E. acervulina alone, suggesting host immune changes induced by berberine. These results suggest that berberine, which exhibits anticoccidial effects, may have therapeutic and/or prophylactic potential in protecting the host from infectious and economic-loss-causing diseases, such as Eimeria infection. Full article

Introduction: Humanitarian Corridors are part of Protected Entry Procedures (PEPs), which allow for the safe and legal arrival in Europe of refugees in need of protection (art. 25 of Regulation (CE) n.810/2009) and were implemented for the first time in Italy in 2015. They represent an alternative to dangerous journeys, and they also bring benefits to local communities. The National Institute for Health, Migration and Poverty (NIHMP) represents the first filter at entry with regard to health needs, as it guarantees an overall individual health assessment to verify the possible presence of ongoing infectious pathologies and of issues that may require management and medical investigation. The aim of this study is to expose the organizational activity and analyze the sociodemographic and health data relating to the Humanitarian Corridors implemented in Italy and in which the NIHMP has participated from 2018 to 2024. Materials and Methods: The organizational lists and health data of each corridor were collected. The analysis was carried out in relation to trend, sociodemographic characteristics of the sample, identification, reception and health. The p-value was considered statistically significant if less than 0.01. In all cases in which the p-value was found to be statistically significant, Cramer’s V was calculated to evaluate the strength of the individual correlation. Results: The NIHMP has participated in 14 Humanitarian Corridors, allowing 1250 refugees to enter Italy; a total of 174 unaccompanied foreign minors (UFMs) arrived, and there were 451 refugees reported as vulnerable (36.1%). Contagious infectious conditions were identified in 223 refugees (17.8%), and other health findings were identified in 414 refugees (33.1%). In the analysis by sex, there are statistically significant differences in the reports of vulnerability and in nationality and education. The inferential analysis carried out by age groups presents statistically significant differences in the reports of vulnerability and in sex and nationality. The analysis relating to the differences by nationality shows statistically significant results in the individual corridors in the reports of vulnerability and in schooling, skin manifestations and infectious diseases and the presence of other health findings. Conclusions: The data reported and analyzed in this work can be considered the first attempt at a complete and detailed analysis regarding the actual implementation and effectiveness of Humanitarian Corridors as programs for legal and safe entry into Italy. The significant gap in research is the reason why there are characteristics of our sample that have no counterpart in the literature. Despite this, for other characteristics, it is possible to find statistical significance and scientific value in line with the data reported both on the websites dedicated to the topic and on the limited scientific literature on the subject. Full article

Background: Malnutrition is increasingly prevalent due to rising life expectancy, oncological cases, and chronic diseases. Early detection is crucial for rehabilitation, complication prevention, and cost reduction. However, nutritional support is often suboptimal. This study aimed to determine malnutrition prevalence and nutritional support status within our hospital. Methods: A point prevalence study was conducted in adult inpatient clinics (excluding pediatrics, infectious diseases, and intensive care) by a 12-member team following ethical approval. NRS-2002 scores, arm/calf circumferences, BMI, and laboratory data (albumin, leukocytes, lymphocytes, neutrophils, hemoglobin, CRP, creatinine) were assessed. Enteral and parenteral nutrition treatments were recorded. Patients with NRS-2002 scores ≥ 3 were classified as at risk of malnutrition. Results: Among 178 patients, 24.7% were at risk of malnutrition. Surgical clinics had a higher malnutrition risk (32.3%) than internal medicine clinics (20.3%). Only 27.1% of at-risk patients received nutritional support (surgical: 19%, internal medicine: 44%). Patients at risk of malnutrition exhibited significantly lower arm circumference (p: 0.000), calf circumference (p: 0.002), lymphocyte counts (p: 0.000), hemoglobin (p: 0.018), albumin (p: 0.001), and BMI (p: 0.038), as well as significantly higher age (p: 0.000) and CRP levels (p: 0.000). Conclusions: Nutritional support remains inadequate despite increased attention to malnutrition. Intensified nutrition education, particularly in surgical inpatient clinics, is needed to improve patient rehabilitation and outcomes. Full article

Overfeeding, currently defined as providing excessive energy and nutrients beyond metabolic requirements, is a common yet often overlooked issue in the intensive care unit (ICU) setting. Understanding the factors contributing to overfeeding and implementing strategies to prevent it is essential for optimizing patient care in the ICU. Several factors contribute to overfeeding in the ICU, including inaccurate estimation of energy requirements, formulaic feeding protocols, and failure to adjust nutritional support based on individual patient needs. Prolonged overfeeding can lead to insulin resistance and hepatic dysfunction, exacerbating glycemic control, increasing the risk of infectious complications, and worsening clinical outcomes. Clinically, overfeeding has been linked to delayed weaning from mechanical ventilation, prolonged ICU stay, and increased mortality rates. Regular review and adjustment of feeding protocols, incorporating advances in enteral and parenteral nutrition strategies, are essential for improving patient outcomes. Clinicians must be proficient in interpreting metabolic data, understanding the principles of energy balance, and implementing appropriate feeding algorithms. Interdisciplinary collaboration among critical care teams, including dieticians, physicians, and nurses, is crucial for ensuring consistent and effective nutritional management. Overfeeding remains a significant concern in the ICU after discharge as well, implying further complications for patient safety and integrity. By understanding the causes, consequences, and strategies for the prevention of overfeeding, healthcare providers can optimize nutrition therapy and mitigate the risk of metabolic complications. Through ongoing education, interdisciplinary collaboration, and evidence-based practice, the ICU community can strive to deliver personalized and precise nutritional support to critically ill patients. Full article

Enteric viruses, such as the Aichi virus (AiV), pose a potential health risk due to their high excretion rates through fecal elimination, limited removal during treatment processes, and prolonged survival, highlighting the need to assess the potential for exposure and disease transmission through sanitation systems. This study investigated the prevalence of AiV at three key stages of sewage treatment in the city of São José do Rio Preto, São Paulo state, Brazil, as well as its viral concentrations, infectious potential, and molecular characterization. The data were also analyzed for potential correlations with reported diarrheal disease cases in the city and the physicochemical properties of sewage. The methodology employed included Nested PCR, qPCR, Sanger Sequencing, and phylogenetic analysis, as well as infectivity testing in cell cultures. The prevalence of AiV throughout the year in raw sewage samples was 90.4%, 78.8% in post-anaerobic biological treatment, and 71.1% in post-chemical treatment, totaling 125 positive samples out of 156, being characterized as AiV genotype A. The virus also demonstrated persistence and infectious potential at all three stages analyzed. The AiV-A mean concentration ranged from 2.05 log¹⁰ to 4.64 GC/mL, 2.31 to 4.72 log¹⁰ GC/mL, and 2.13 to 2.85 log¹⁰ GC/mL for the same treatment stages, respectively. A significant difference (p ≤ 0.05) suggests higher viral concentrations in summer at the three sewage process points analyzed, while lower viral concentrations were observed in post-chemical treatment samples (p ≤ 0.01). Additionally, no statistically significant relationship was observed between the virus occurrence in samples and cases of acute diarrheal diseases in the city. In conclusion, this study highlights that much remains to be understood about AiV while providing valuable insights into the relationship between AiV, environmental factors, and public health. Full article

Background/Objectives: Porcine epidemic diarrhea (PED) is a highly contagious enteric infectious disease that causes severe morbidity and mortality in piglets, posing significant economic losses to the swine industry worldwide. Oral vaccines based on Lactococcus lactis offer a promising approach due to their safety and genetic manipulability. This study aims to develop and evaluate an oral L. lactis-based vaccine expressing the full-length PEDV S protein. Methods: A recombinant L. lactis strain expressing the PEDV S protein was constructed and encapsulated in alginate–chitosan microcapsules. Vaccine stability was tested in simulated digestive fluids, and mice were orally immunized. Immune responses were evaluated by measuring specific antibodies, cytokines, and lymphocyte proliferation. Results: The recombinant L. lactis NZ3900/pNZ8149-S strain successfully expressed the full-length PEDV S protein and maintained stable plasmid inheritance. Oral immunization in mice induced detectable PEDV-specific immune responses. Both encapsulated and non-encapsulated vaccines stimulated the production of IgG and sIgA antibodies, as well as cytokines associated with Th1 and Th2 responses. Notably, encapsulation with alginate–chitosan significantly enhanced bacterial survival in digestive conditions and further amplified immune responses, including higher antibody titers, elevated levels of IFN-γ, IL-4, and IL-10, and greater lymphocyte proliferation, indicating improved immune memory. Conclusions: The oral L. lactis NZ3900/pNZ8149-S vaccine expressing the PEDV S protein effectively induced systemic and mucosal immunity in mice. Encapsulation with alginate–chitosan further enhanced its immunogenicity and stability in gastrointestinal conditions. These results suggest that both the engineered L. lactis strain and the encapsulation strategy contribute to the development of a promising oral vaccine platform for controlling PEDV in swine populations. Full article

Poor health and increased susceptibility to infectious diseases are among the main sources of economic losses in the pig industry worldwide, and they also serve as indicators of compromised animal welfare. However, there is limited information on long-lasting biomarkers of poor health and common infections experienced by piglets early in life. Hair cortisol, dehydroepiandrosterone sulfate (DHEA(S)), and their ratio have been proposed as components of the mammalian stress response due to the activation of the hypothalamus–pituitary–adrenal axis and were investigated in this study using 30 batches of pigs from 16 farms. The research hypothesis was that batches of piglets experiencing clinical syndromes (as indicated by enteric, neurological, cutaneous, and locomotor scores) during suckling would exhibit a different pattern of resilience and allostatic load later in life compared to healthy ones. Hair from 25 gilts per batch were collected at either 3.5 or 9 months of age, and hormone extraction was subsequently performed. The farm of origin and the age of the animals significantly influenced hormone concentrations. Moreover, batches affected by enteric disease showed lower DHEA(S) levels (p < 0.0001; 15.89 vs. 23.51 pg/mg) and higher cortisol/DHEA(S) ratio (p < 0.0001; 82.83 vs. 55.02) than healthy batches. Similar results were observed in batches with a neurological syndrome (DHEA(S): p < 0.0001; 12.91 vs. 19.43; cortisol/DHEA(S) ratio: p < 0.0001; 97.15 vs. 70.26 pg/mg). These results suggest that pig hair biomarkers carry an intrinsic and temporally stable signal related to early life health status. Full article

The interplay between nutrition and infectious diseases has been a central theme in health sciences for the last decades due to its great impact on the pediatric population, especially in immunocompromised patients and critically ill children. As conventional treatment and the development of antimicrobials for most infections standard treatment is either limited or not possible, alternative treatment options should be explored. Recent research shows that early enteral nutrition and nutritional supplements (such as probiotics and symbiotics) could have a pivotal role in promoting a healthy microbiome and subsequently preventing and improving outcomes for certain pediatric infectious diseases. However, understanding the specific mechanism of action and tailoring nutritional interventions remains a significant challenge. The optimal dose range for different probiotic strains and prebiotics and the most effective combination for each treatment indication needs further investigation and is yet to be defined. Additionally, in the era of personalized medicine, goal- and patient-directed treatment are key to optimizing and improving outcomes and minimizing potential complications and side effects, especially in complex and immunocompromised patients. The main objectives of this narrative review are 1. to explore the relationship and the complex interactions between microbiota and the human immune system; 2. to describe the influence of nutrition on infectious diseases; 3. to evaluate the impact of supplementation with probiotics and symbiotics in the prevention and treatment of the most relevant infections in children; and 4. to identify knowledge gaps and potential research priorities regarding the use of these supplements in pediatric patients. Full article

Bivalve mollusks face a crisis due to infectious diseases, resulting in high mortality and economic losses. The need for continuous monitoring to prevent contamination from sewage and rainwater in aquaculture is evident. The recent mass mortality of scallops in Ilha Grande Bay (IGB), Rio de Janeiro’s largest scallop producer, due to environmental contaminants underscores the need for further research. This study aims to investigate the recent collapse of the scallop population and assess the human impact by analyzing the circulation of pathogens. Materials and Methods: Mollusks were collected from three sites in Ilha Grande Bay (IGB), a region known for its significant scallop production, and from scallop farms in Angra dos Reis, RJ. A total of 216 gill and adductor tissue samples from lion’s foot scallops were analyzed. Bacterial contamination was identified using MALDI-TOF, while antimicrobial susceptibility and carbapenem production were assessed via disk diffusion tests. Results: Mollusks were contaminated with V. alginolyticus, V. fluvialis, V. harveyi, Pseudomonas putida, and Pseudomonas monteilii. All isolates were sensitive to meropenem, but P. putida showed higher resistance to ciprofloxacin. Conclusions: The presence of these pathogenic and resistant bacteria in scallop adductor tissues is a concern for the aquaculture industry and a significant public health risk. The potential for these bacteria to enter the human food chain through consuming contaminated seafood or recreational activities such as bathing is a serious issue that needs to be addressed. Full article

Botulinum neurotoxins (BoNTs) are the most potent toxins on Earth and are classified as Category A biological agents. BoNTs lead to paralysis in humans and cause botulism. Antibody therapeutics can effectively treat toxin-mediated infectious diseases. In this study, we generated a pharmaceutical humanized monoclonal antibody (HZ45 mAb) to prevent or treat botulism. HZ45 binds to the heavy chain receptor (HCR) domain of the toxin, preventing the toxin from entering the cell. The mAb was produced using hybridoma technology and phage display. We evaluated HZ45 mAb for the neutralization of BoNT serotype A (BoNT/A) in mice and rabbits. The survival results showed that pretreatment with HZ45 mAb provided 100% protection at a dose of 0.1 mg per mouse against a maximum of 100 LD₅₀ of BoNT/A. To assess the therapeutic efficacy of HZ45 mAb in New Zealand white rabbits (NZWs), a 5 mg dose was administered 4 or 8 h after challenge with 10 LD₅₀. The results indicated that 5 mg of HZ45 could treat the NZWs within 8 h after exposure to 10 LD₅₀ botulinum. Consequently, in an in vivo context, including mice and rabbits, HZ45 mAb could protect against botulinum type A intoxication. Full article

Background/Objectives: Feline coronavirus (FCoV) belongs to the family Coronaviridae and includes two pathotypes, the less virulent feline enteric coronavirus (FECV), which replicates in the enteric epithelial cells, and feline infectious peritonitis virus (FIPV), which is more virulent, replicates efficiently within monocytes/macrophages with systemic involvement and may cause feline infectious peritonitis (FIP), a progressive and often fatal disease. The diagnosis of FIP is complex and requires different examinations. Among serological tests, the indirect immunofluorescent antibody test (IFAT), considered the gold standard, and the enzyme-linked immunosorbent assay (ELISA) are the most widely used to detect FCoV antibodies. The aim of this work was the development of FCoVCHECK Ab ELISA, a new rapid indirect test for the detection of FCoV antibodies in feline serum/plasma samples. Methods: FCoVCHECK Ab ELISA was developed after a meticulous set-up and cut-off analysis through several methods, including the Youden’s index and ROC curve, to achieve the best test performance. It was validated by testing 110 feline sera (62 positives and 48 negatives) against the reference IFAT and compared with two other rapid ELISA tests, INgezim Corona Felino (Gold Standard Diagnostics) and ImmunoComb Feline Coronavirus (FCoV) [FIP] Antibody Test Kit (Biogal). Conclusions: FCoVCHECK Ab ELISA agreed with IFAT at 96.4% (93.5% sensitivity, 95% confidence interval (CI): 83.5–97.9%; 100% specificity, 95% CI: 90.8–100%), with ImmunoComb FCoV at 93.6% and with INgezim Corona Felino at 82.7%. Intra- and inter-assay accuracy and precision gave coefficients of variation lower than 20%. Compared to IFAT, the new assay correctly identifies positive and negative samples with a good correlation, and, in addition, it is simpler, faster and provides a less subjective reading of the results. Full article