Search Results (1,663)

Background/Objectives: To overcome bottlenecks in the application of Macleaya cordata extract (MCE) in veterinary traditional Chinese medicine, such as low bioavailability of its active ingredients, gastrointestinal irritation, and muscular toxicity, this study aimed to develop a liposomal nano-delivery system loaded with MCE (MCE-Lips) to achieve the core objective of “enhancing efficacy and reducing toxicity” and to explore its potential application and mechanism of action in treating bacterial diarrhea. Methods: MCE-Lips were prepared using the thin-film dispersion method, and their physicochemical properties—particle size, encapsulation efficiency, and drug loading capacity—were characterized. In vitro, cytotoxicity against skeletal muscle cells and NCM460 intestinal epithelial cells was evaluated using the CCK-8 assay. The release of lactate dehydrogenase (LDH) from skeletal muscle cells was measured with an LDH assay kit. The expression levels of inflammatory factors (TNF-α, IL-6, and IL-1β) in both cell types were determined through ELISA. A fluorescent probe was employed to assess cell membrane integrity. The effect of MCE-Lips on the expression of tight junction proteins (ZO-1, Occludin, and Claudin-5) was evaluated via immunofluorescence. Acute toxicity was examined through H&E staining. A bacterial diarrhea model was established using Escherichia coli in mice, and comprehensive safety and efficacy were assessed through hematological tests and gastrointestinal motility evaluation. Finally, untargeted metabolomics and 16S rRNA sequencing were utilized to investigate the underlying mechanisms of action. Results: The prepared MCE-Lips had an average particle size of 86.49 nm and a high encapsulation efficiency of 89.07%. In vitro experiments demonstrated that MCE-Lips significantly alleviated skeletal muscle cell damage, reduced LDH release (p < 0.05), and effectively inhibited the expression of inflammatory factors IL-6, TNF-α, and IL-1β (p < 0.05). In NCM460 cells, MCE-Lips exhibited a more pronounced inhibitory effect on LPS-induced release of TNF-α (p < 0.01), IL-6 (p < 0.0001), and IL-1β (p < 0.0001) and enhanced intestinal barrier function by upregulating the expression of tight junction proteins ZO-1 (p < 0.001), Occludin (p < 0.01), and Claudin-5 (p < 0.01). In the bacterial diarrhea model, MCE-Lips showed excellent anti-diarrheal efficacy (p < 0.01). Hematological analysis indicated no systemic toxicity. At the endocrine level, the high-dose group significantly reduced motilin (MTL) levels (p < 0.01), which slowed intestinal motility and prolonged chyme retention, thereby alleviating diarrhea symptoms. Mechanistic studies revealed that it acts by regulating the intestinal metabolic profile and microbial community structure, with Desulfovibrio, Enterococcus, and Streptococcus identified as key characteristic differential genera. Conclusions: For the first time, an MCE liposome nanoparticle system was constructed, and untargeted metabolomics combined with 16S rRNA sequencing were employed to elucidate its anti-diarrheal mechanism. MCE-Lips exerts excellent antibacterial diarrhea effects through multiple mechanisms, including direct cytoprotection and anti-inflammatory action, enhancement of the intestinal barrier, regulation of gut function, and remodeling of the gut microecology. This work provides a novel paradigm for plant-derived nano-anti-diarrheal agents. The systematic evaluation of the pharmacodynamics of MCE-Lips in a piglet bacterial diarrhea model will lay a solid foundation for its eventual market application. Full article

Background/objectives: A rise in infections associated with carbapenem-resistant Providencia species (CRPS) has been observed worldwide. This study presents a genomic analysis of CRPS isolates from four hospitals in Croatia and the outpatient setting, in order to determine the extent of the spread of CRPS in Croatia. In the present study, we applied a combination of phenotypic characterization and molecular analysis of resistance traits to determine the mechanisms and the routes of spread of CRPS. Material and methods: The antibiotic susceptibility testing was performed using disk-diffusion and broth dilution methods. The nature of extended-spectrum β-lactamases (ESBLs), carbapenemases, and fluoroquinolone resistance determinants was investigated by polymerase chain reaction (PCR). In order to obtain an insight into the whole resistome, selected isolates were subjected to the Interarray Genotyping Kit CarbaResist and whole genome sequencing (WGS). Results: In total, 30 isolates were collected from four centers, located in different geographic regions of Croatia. There was uniform resistance to piperacillin-tazobactam, cefuroxime, expanded-spectrum cephalosporins (ESCs), imipenem, ertapenem, meropenem, and ciprofloxacin. Immunochromatographic testing and PCR revealed OXA-48 and NDM carbapenemase in 15 isolates, respectively. Phenotypic tests for ESBLs were positive in all OXA-48 and one NDM-positive organism (16 isolates). The isolates were categorized as extensively drug-resistant (XDR). OXA-48-producing isolates were susceptible only to ceftazidime-avibactam, whereas NDM producers were susceptible to cefiderocol and, in the majority of cases, also to amikacin. WGS identified a plethora of genes encoding resistance to aminoglycosides, such as aadA1 and aadA2, (aph(3″)-Ib and aph(6)-Id, sulfonamides sul1 and sul2, trimethoprim dfrA1, dfrA10, and dfrA12, tetracyclines tet(A) and tet(B), and chloramphenicol catA3 and catA5. Conclusions: Providencia spp., in spite of being a rare pathogen, should be included in the surveillance studies across the medical centers in Croatia. Full article

Background/Objectives: Taiwan cinnamon leaves have been reported to be effective in improving chronic diseases. Herein, cinnamon leaf extract (CLE) and nanoemulsion (CLEN) were prepared to explore their effects in improving sexual dysfunction in rats. Methods: Following extraction with 80% ethanol and analysis by UPLC-MS/MS, CLEN was prepared using an optimal ratio of soybean oil, lecithin, Tween 80, deionized water, and CLE. A total of 48 male rats and 48 female rats were used, with the former being induced with erectile dysfunction, followed by treatment with CLEN or CLE at two doses (100 mg/kg and 50 mg/kg) for 4 weeks. After conducting the penile reflex test, male rats were paired with female rats for measurement of sexual behavior and ICP/MAP. Following sacrifice, α-SMA, nNOS, and β-III tubulin expression areas were measured by histochemical analyses; SMC/collagen ratio by Masson’s trichrome staining; and NO, cGMP, and PDE5 levels by ELISA kits. Results: CLEN was more effective than CLE in increasing intromission frequency, decreasing intromission and ejaculation latency, and recovering erectile response for improving copulatory and ejaculatory performances. A higher maximum ICP/MAP ratio was shown for CLEN through elevation of neurovascular function and erectile capacity. Additionally, CLEN efficiently reduced fibrosis, enhanced neuronal marker expression, and increased the SMC/collagen ratio, leading to penile tissue protection and neural regeneration. Both treatments showed elevated levels of NO and cGMP with a reduction in PDE5, probably through modulation of the NO-cGMP signaling pathway. Conclusions: CLEN was more effective than CLE in restoring erectile function in rats. Some more clinical trials are needed to verify this finding. Full article

Background/Objectives: Dental anxiety is a common clinical problem that negatively affects patient cooperation, treatment acceptance, and physiological stability during dental procedures. This randomized controlled clinical trial study aimed to evaluate the effectiveness of music-based interventions in reducing dental anxiety and stress responses during restorative dental treatment. The null hypothesis was that music exposure would not result in significant differences in anxiety levels or physiological stress parameters compared with standard dental care. Methods: Seventy-five patients with moderate to high pre-treatment dental anxiety (MDAS ≥10) were randomly assigned to three groups: classical music, Turkish music, and control (no music) (n = 25 per group). Anxiety levels were assessed using the Modified Dental Anxiety Scale (MDAS). Restorations were performed using a standardized adhesive protocol. Physiological parameters, including systolic and diastolic blood pressure (SBP, DBP), heart rate (HR), and oxygen saturation (SpO₂), as well as salivary cortisol and alpha-amylase levels, were measured before and after restorative treatment. Salivary cortisol and amylase levels were measured using a Human ELISA Kit. Statistical analysis was performed using paired t-tests and one-way ANOVA with Tukey’s post hoc test (p < 0.05). Results: Both music groups showed significant reductions in SBP, DBP, HR, cortisol, amylase, and MDAS scores compared to the control group (p < 0.05). Oxygen saturation increased significantly in the music groups, whereas it decreased significantly in the control group. There were no significant differences between classical and Turkish music regarding their anxiety-reducing effects. Conclusions: Music-based interventions effectively reduce dental anxiety and physiological stress during restorative dental procedures. This study is novel in simultaneously evaluating subjective anxiety scores and multiple physiological and biochemical stress markers in adult patients undergoing restorative treatment, supporting music as a simple and non-invasive adjunct in clinical dentistry. Full article

Commercial immunoassay-based test kits are widely used for rapid screening of deoxynivalenol (DON) in maize; however, inconsistent results are frequently observed under commercial testing conditions. This study evaluated two distinct contributors to such variability: analytical cross-reactivity of commercial DON immunoassays and between-site variability arising from routine grain elevator testing practices. Under controlled laboratory conditions, all kits accurately measured DON but responded differently to co-occurring DON derivatives. In naturally contaminated maize, immunoassay results reflected the combined presence of DON and co-occurring derivatives, consistent with differences in antibody specificity. An interlaboratory comparison involving multiple grain elevators analyzing identical blinded samples demonstrated substantial between-site variability in reported DON concentrations, with about 16% of results deviating by more than ±20% from the LC–MS/MS reference value. Collectively, these findings show that inconsistent DON test outcomes arise from the combined effects of antibody cross-reactivity and site-specific testing variability, rather than from any unreliability of the analytical methods themselves. This finding highlights the importance of interpreting rapid DON measurements considering these factors. Full article

The drug-encapsulated hybrid hydrogels possessed several expected properties, including porous microstructure, conductivity, adhesive strength, antioxidant activity, antibacterial properties, and cytocompatibility, and have great potential in biomedical applications, such as skin wound hydrogel dressings and bio-adhesives. In this paper, the quercetin-loaded hybrid hydrogels (SSA-QRs) were fabricated using silk fibroin (SF), alginate, and silver-doped reduced graphene oxide (Ag@rGO) nanosheets, incorporating quercetin-encapsulated PF-127 (PF127-QR) micelles. Scanning electron microscopy (SEM) images confirmed that the fabricated hybrid hydrogels possessed an interconnected porous microstructure. The mechanical properties of hydrogels could be regulated by adjusting the content of incorporated Ag@rGO nanosheets and PF127-QR micelles. Furthermore, the obtained SSA-QR hydrogels displayed the expected swelling properties, and the swelling rates could reach 1200–1700% in 120 min, in the equilibrium state. The fabricated SSA-QR hydrogels possessed apparent conductivity and self-healing ability. In addition, SSA-QR hydrogels exhibited strong adhesive performance on the surface of different materials, including skin, metal, wood, plastic, and glass. The typical antibacterial testing using Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli) confirmed the excellent antibacterial activities of SSA-QR hydrogels. Moreover, SSA-QR hydrogels displayed good antioxidant ability and intracellular ROS scavenging ability. However, the increased content of Ag@rGO nanosheets could cause a great increase in the hemolysis ratio for SSA-QR hydrogels. Fluorescent images, cell counting kit-8 (CCK-8) assay, and cell scratch testing confirmed their excellent cytocompatibility and cell pro-migration ability. The available results demonstrated a facile strategy to prepare the quercetin-loaded hydrogel for applications of wound hydrogel dressing and bio-adhesives. Full article

Toxoplasma gondii is a globally distributed zoonotic parasite that infects a wide range of warm-blooded animals and poses serious threats to many endangered species. Effective serological detection is critical for disease surveillance and control, yet the accuracy of commercial assays varies significantly across host species. Validated serological methods for the Chinese red panda (Ailurus styani) remain scarce, limiting effective monitoring and conservation efforts. A total of 57 serum samples were collected from captive red pandas. Three commercial serological kits—the modified agglutination test (MAT), indirect hemagglutination assay (IHA), and enzyme-linked immunosorbent assay (ELISA)—were used to detect T. gondii antibodies. Statistical analyses included agreement assessment (Cohen’s kappa), McNemar’s test, and receiver operating characteristic (ROC) curve analysis, with MAT as the reference standard. MAT detected T. gondii antibodies in 36.84% (21/57; 95% CI: 24.32–49.36) of samples, while ELISA identified 42.11% (24/57; 95% CI: 29.29–54.92). IHA failed to detect any positive samples (0%). MAT and ELISA showed almost perfect agreement (κ = 0.817; 95% CI: 0.66–0.97) with no significant difference (p > 0.05). Compared to MAT, ELISA exhibited 95.2% sensitivity and 88.9% specificity. The combined MAT/ELISA approach with retesting of discordant samples yielded a final seroprevalence of 35.09% (20/57), with 8.77% (5/57) indeterminate results. MAT and ELISA are reliable methods for serological surveillance of T. gondii in red pandas, while IHA is unsuitable. We recommend MAT for initial screening followed by ELISA confirmation. The high seroprevalence highlights the need for integrated control strategies, including management of stray felids and rodents in captive environments to reduce transmission risks. Full article

In recent decades, technological advances have fostered new therapeutic approaches for children with neurodevelopmental disorders, particularly Autism Spectrum Disorder. Educational robotics has emerged as a promising resource for acquiring social skills, recognizing emotions, and developing theory of mind. However, there is still a need to understand which dimensions are most susceptible to this specific intervention and how its impact differs based on individual profiles. This study analyzes the effect of a therapeutic intervention based on Educational Robotics on Social Skills, Emotional Recognition, and Theory of Mind in two students diagnosed with Autism Spectrum Disorder. The intervention was structured in seven sessions using the ANDY^® kit. Tests from the NEPSY-II battery and an observational rubric of Social Skills recorded session by session were applied. Both participants showed significant improvements in Social Skills, especially in rules of courtesy, nonverbal communication, and conversational interaction. Regarding Emotional Recognition, one participant improved in identifying highly affective emotions, while the other showed more modest progress. Theory of Mind showed progress in only one of the participants. Adherence was high, although a slight decrease in motivation was identified in the last sessions. These results suggest that Educational Robotics, when applied within a structured therapeutic framework, can effectively foster socioemotional development in children with Neurodevelopmental Disorders. However, variability across domains highlights the importance of tailoring interventions to individual profiles and complementing them with strategies that support the transfer of learning to natural contexts. Full article

Background: Pancreatic cancer (PC) and colorectal cancer (CRC) are among the most lethal malignancies, with growing evidence pointing to the gut microbiota’s role in their progression. This study aimed to explore the anticancer potential of two microbiota-derived postbiotics, N-acetylcysteine (NAC) and tetrahydro β-carboline carboxylic acid (THC), in targeting some hallmark traits of PC and CRC, both as standalone agents and in combination with standard chemotherapeutics (gemcitabine for PC and 5-fluorouracil (5-FU) for CRC). Methods: Cell viability assays and IC₅₀ determination was assessed using either the Muse™ Count & Viability Kit or the Sulforhodamine B assay; cell death was determined by Annexin V/Propidium Iodide and cell cycle assessed by Propidium Iodide was analyzed by flow cytometry. Results: Here, we found that NAC selectively reduced the viability of PC cells BxPC-3 without triggering apoptosis, while effectively inducing apoptosis in PC cells Panc-1 and in CRC cell lines. THC exhibited stronger anticancer activity, inhibiting proliferation and promoting apoptosis in all tested PC and CRC cells, even at lower concentrations. Combination treatments yielded promising enhancement effects. NAC enhanced the cytotoxicity of gemcitabine in Panc-1 cells through increased apoptosis. NAC, when combined with 5-FU, also increased apoptosis of CRC cells. THC further potentiated gemcitabine’s impact on Panc-1 cells by increasing apoptosis and by inducing cell cycle changes in BxPC-3. In the CRC model, THC co-treatment with 5-FU reduced cell viability and increased apoptosis in all cells. Conclusions: These findings provide preliminary in vitro evidence supporting the potential of integrating microbiota-derived postbiotics with conventional chemotherapy both in PC and CRC. Full article

As an emerging threat to global food security, wheat blast necessitates the development of a rapid and field-deployable detection system to facilitate early diagnosis, enable effective management, and prevent its further spread to new regions. In this study, we aimed to validate and improve a Recombinase Polymerase Amplification coupled with PCRD lateral flow detection (RPA-PCRD strip assay) kit for the rapid and specific identification of Magnaporthe oryzae pathotype Triticum (MoT) in field samples. The assay demonstrated exceptional sensitivity, detecting as low as 10 pg/µL of target DNA, and exhibited no cross-reactivity with M. oryzae Oryzae (MoO) isolates and other major fungal phytopathogens under the genera of Fusarium, Bipolaris, Colletotrichum, and Botrydiplodia. The method successfully detected MoT in wheat leaves as early as 4 days post-infection (DPI), and in infected spikes, seeds, and alternate hosts. Furthermore, by combining a simplified polyethylene glycol-NaOH method for extracting DNA from plant samples, the entire RPA-PCRD strip assay enabled the detection of MoT within 30 min with no specialized equipment and high technical skills at ambient temperature (37–39 °C). When applied to field samples, it successfully detected MoT in naturally infected diseased wheat plants from seven different fields in a wheat blast hotspot district, Meherpur, Bangladesh. Training 52 diverse stakeholders validated the kit’s field readiness, with 88% of trainees endorsing its user-friendly design. This method offers a practical, low-cost, and portable point-of-care diagnostic tool suitable for on-site genomic surveillance, integrated management, seed health testing, and quarantine screening of wheat blast in resource-limited settings. Furthermore, the RPA-PCRD platform serves as an early warning modular diagnostic template that can be readily adapted to detect a wide array of phytopathogens by integrating target-specific genomic primers. Full article

Reticuloendotheliosis virus (REV) is an immunosuppressive virus in poultry that can cause acute reticular neoplasms, chronic lymphoid tumors, stunting syndrome, and secondary infections. In many countries, the lack of effective vaccines has resulted in a high prevalence of REV infections and substantial economic losses. Enzyme-linked immunosorbent assay (ELISA)-based antibody detection is an important tool for monitoring the REV prevalence in poultry farms. ELISA coating antigens generally consist of either whole virus or viral protein; however, most commercially available REV antibody ELISA detection kits use whole virus as the coating antigen, which limits their applicability in certain diagnostic and research settings. In this study, the gp90 protein from a dominant REV strain was expressed and purified using 293F suspension cell eukaryotic expression system. Using recombinant gp90 protein as the coating antigen, an indirect ELISA for detecting gp90 antibodies (gp90-ELISA) was developed. After optimization, the optimal conditions were as follows: coating antigen concentration of 4 µg/mL with overnight incubation at 4 °C; blocking with 5% skim milk at 37 °C for 1.5 h; serum dilution of 1:200 with incubation at 37 °C for 45 min; secondary antibody dilution of 1:1000 with incubation at 37 °C for 30 min; and color development using TMB substrate at room temperature in the dark for 10 min. The cut-off value was defined as an OD₄₅₀ ≥ 0.22 for positive samples and <0.22 for negative samples. The developed gp90-ELISA specifically detected REV-positive sera at a maximum serum dilution ratio of 1:3200. Intra- and inter-assay variation coefficients were ≤10%, indicating that the gp90-ELISA had good specificity, sensitivity, and reproducibility. Laboratory serum testing showed that the gp90-ELISA successfully detected sera from chickens immunized with the gp90 protein or infected with REV. Furthermore, analysis of clinical serum samples demonstrated 100% concordance between the gp90-ELISA results and a commercial whole-virus-coated ELISA kit. These results indicate that the gp90-ELISA is a reliable supplementary method to whole-virus-coated ELISA and has potential utility in disease surveillance and evaluation of immune responses. Full article

Pathogens such as Toxoplasma gondii, lentiviruses (e.g., CAE), Hypoderma spp., Neospora caninum, Mycoplasma spp., and pestiviruses are important for goat farming in Lithuania; however, data on their prevalence remain limited. To address this gap, a multi-pathogen study was conducted between 2021 and 2024 using selected ELISA kits (ID.vet, Innovative Diagnostics, France). A total of 380 blood samples were collected from 30 goat herds across different regions of Lithuania; the sample size varied depending on the pathogen. Serum samples were tested for antibodies, and seroprevalence was calculated for each pathogen. The highest seroprevalence was detected for T. gondii (38.9%, 143/368) and CAE virus (19.5%, 74/380). Antibodies to Mycoplasma spp. (0.3%, 1/368), Hypoderma spp. (3.8%, 7/184), and N. caninum (0.5%, 2/368) were detected only sporadically, while no antibodies to Border disease virus or Q fever were identified. Mixed infections were found in 7.6% of samples. Chi-square analysis showed that co-infections with toxoplasmosis and CAE occurred more frequently than expected (χ² = 19.05, p < 0.001). Herd size was significantly associated only with CAE seroprevalence (χ² = 7.913, df = 1, p < 0.05). Overall, toxoplasmosis and CAE were identified as the most epidemiologically relevant infections in the Lithuanian goat population. Full article

Soybean wax is a sustainable alternative to synthetic polymeric coatings in packaging due to its renewable, environmentally benign, and hydrophobic properties. In order to be effectively applied, however, soybean wax must be emulsified in water. The present work compares two stabilization approaches for soybean wax emulsions: a conventional surfactant-based emulsion (SE) using a mixture of nonionic surfactants (Span-80 and Tween-80), and a Pickering emulsion (PE) using cellulose nanocrystals combined with sodium alginate (CNC-SA) as an anionic stabilizer. The SE produced stable emulsions at 6 wt% Span-80/Tween-80 (at a HLB_mix value of 10) with a mean droplet size of 449 nm but limited storage stability (approximately 7 days under ambient conditions), while the PE achieved superior stability (approximately 1 month) at 1 wt% CNC-SA with a mean droplet size of 740 nm. The stabilized SE and PE were subsequently applied as coatings on three different types of paper substrates: northern bleached kraft (NBK) paper, copy paper, and cellulose nanofiber (CNF)-coated NBK paper. When applied to northern bleached kraft (NBK) paper, the SE coatings provided minimal improvements in barrier performance. The Cobb 60 value decreased slightly from 125 g/m² (control-no coating) to 86 g/m², indicating a negligible water barrier with immediate water absorption upon contact. In contrast, the Cobb 60 value of the PE-coated NBK paper decreased markedly from 125 g/m² to 39 g/m², confirming that the PE coating substantially enhances water resistance. The SE coating displayed a significant loss of water contact angle (WCA) from 85° to 0° within 20 s, showing limited water holdout capacity, whereas PE-coated NBK paper demonstrated strong water holdout, with the WCA decreasing only from 94° to 85° over 5 min. The SE coating achieved only a 14% reduction in water vapor transmission rate (WVTR), while the PE coating provided a greater reduction of 30%. In terms of oil resistance, both emulsion systems significantly enhanced the kit rating of the papers tested, e.g., from kit number 0 to 6–9 (paper dependent). The SE coating, however, experienced a substantial reduction in barrier integrity after folding, while the PE coating largely retained its oil barrier properties. Furthermore, the SE coating reduced the tensile strength of NBK paper by 41%, whereas the PE coating reduced it by only 7%. Overall, the comparative findings indicate that although the SE generated a smaller mean particle size, it offered minimal improvement in the water and oil barrier performance of paper and had a limited storage life. In contrast, the PE generated a larger mean particle size, but provided substantially greater water and oil resistance, and enhanced mechanical strength retention. In addition, the PE displayed an effective storage life of at least one month. The Pickering emulsion, formulated with all biologically derived components, therefore represents a viable, sustainable, bio-based alternative to synthetic polymeric coatings for packaging applications. Full article

Monitoring of antimicrobial resistance (AMR) in the environment is needed, but standardized methods are lacking. This study evaluated a cost-efficient, kit-based approach for detecting antibiotic-resistant Escherichia coli (AR-E. coli) and predicting multidrug-resistant E. coli (MDR-E. coli) in concentrated animal feeding operations (CAFO)-impacted surface waters in Tulare, California, and Clayton, Michigan. Eighteen Tulare and six Clayton samples were analyzed using modified IDEXX Colilert-18 kits with selective antibiotics. In Round 1 of the study, 702 isolates were tested against 12 antibiotics using disk diffusion to identify ideal additions. No and low resistance to AMP was seen at the unimpacted and less-impacted sites, respectively, while up to 34% of isolates at impacted sites were resistant to AMP. The percentage of isolates resistant to erythromycin was 16% or lower at less-impacted sites, but it ranged up to 64% at impacted sites. In Round 2, 1002 isolates (1704 total) were characterized to compare modified IDEXX results with culture-based results. Resistance to ampicillin (AMP-E. coli-col) detected by IDEXX strongly correlated with MDR prevalence in Clayton (R² = 0.71, p = 0.00197), while extended-spectrum beta-lactamase-producing E. coli (ESBL-E. coli-col) correlated with MDR prevalence in Tulare (R² = 0.70, p < 0.0001). This observation appears to be site-specific. Findings suggest both IDEXX- and culture-based methods can serve as AMR screening tools at CAFO-impacted sites. This work supports community-based monitoring and informs standardized approaches for tracking AMR in inland waters. Full article

Background: Bordetella pertussis causes pertussis, a respiratory infection with whooping cough. Despite a high vaccine coverage, pertussis resurged post-COVID-19 pandemic. Meanwhile, isolates resistant to macrolides—the first-line therapy—have increased in several countries, including Japan. Culturing B. pertussis and detecting resistance are difficult; reports remain limited in Japan. Methods: From March to August 2025, we collected nasopharyngeal samples from children aged 0–15 years with suspected pertussis at six Japanese clinics. Pediatricians obtained swabs and tested them using gene-amplification kits (e.g., BioFire^® SpotFire^® in four clinics, LAMP Pertussis Detection^® in two clinics). B. pertussis was confirmed by PCR; isolates were sequenced to identify macrolide-resistant mutations. Results: Samples were taken from 54 children, the number of boys and girls was 34 and 20, and their median age was 12 years old. Among 54 B. pertussis isolates, 43/52 (82.7%) sequenced strains harbored the A2047G mutation associated with macrolide resistance. Resistance rates at each clinic varied from 40% to 96%. Conclusions: These findings indicate a post-pandemic rise in macrolide-resistant B. pertussis in Japan. Ongoing resistance surveillance is essential, and repurposing residual clinical samples after routine testing is useful given culture and detection challenges. Full article