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Online food delivery platforms increasingly shape food environments and dietary choices, yet it remains unclear whether platform-defined "healthy" outlet categories align with evidence-informed assessments. This study assessed the healthiness of food outlets labelled as "healthy" on a leading online food delivery platform in Victoria, Australia, using food environment classification tools. Cross-sectional study using web-scraped outlet-level data. Outlets labelled "healthy" were assigned to one of 36 predefined outlet types and classified using the DIGIASSESS index, an expert-informed food environment scoring tool. A supplementary menu-based sensitivity analysis was conducted in a stratified random 10% subsample of outlets. A leading online food delivery platform in Victoria, Australia. We identified 12,938 unique food outlets, of which 1,408 (9.2%) were labelled "healthy" by the platform and included in the primary analysis. A stratified random subsample of 166 outlets underwent menu-level review. "Healthy"-labelled outlets were most commonly Independent-Takeaway (11.8%), Independent-Cereal-Based Café Meals (9.7%), and Service Station Convenience Stores (7.4%). Using DIGIASSESS, most "healthy"-labelled outlets were reclassified as "less healthy" (n = 1,123; 79.7%) or "unhealthy" (n = 180; 12.8%), with 106 (7.5%) classified as "healthy." The supplementary menu-based analysis showed similar classifications to DIGIASSESS (>98% agreement). On this leading Australian online food delivery platform, the "healthy" outlet category did not align with expert-informed assessments of outlet-type healthiness. Such misalignment risks misleading consumers and highlights the need for transparent, standardised criteria governing health-related outlet categories in digital food environments.

There is growing concern about the impact of social media on mental health. Understanding the psychological processes underlying social media use could lead to greater insight. To investigate individual differences in the effect of social media likes on posting behavior and their association with and specificity to depressive psychopathology. This cross-sectional study used longitudinal Twitter behavior to test the association between depressive and other forms of psychopathology and the tendency to be reinforced by likes. The study involved passively scraped Twitter data (dataset 1) or participants recruited online through Twitter ads (dataset 2) and a recruitment platform (dataset 3). In total, the 3 datasets contained over 17 million posts from 7736 Twitter users. These data were analyzed from March 2023 through January 2026. Reinforcement tendency was estimated as the association between previous-day likes and current-day posting. The depression measure varied between datasets: self-disclosure in dataset 1, past 4-week depression severity in dataset 2, and a validated questionnaire in dataset 3. Dataset 3 also included other psychiatric measures used to estimate scores on transdiagnostic symptom scores. Dataset 1 comprised 1045 users who reported a depression diagnosis on Twitter, and 5001 randomly sampled users. Dataset 2 comprised 601 users (mean [range] age, 45.2 years [18-78]; 251 [41.7%] women, 333 [55.5%] men, and 17 [2.8%] individuals with other gender identities). Dataset 3 comprised 1089 users (mean [range] age, 30.8 years [18-68]; 703 [64.6%] women, 358 [32.8%] men, and 28 [2.6%] individuals with other gender identities). Individuals with a depression diagnosis or higher symptoms of depression tended more to be reinforced by likes in all datasets (dataset 1: β = .013; P = .002; dataset 2: β = .026; P = .03; dataset 3 [preregistered]: β = .008, P = .02), despite substantial differences between datasets in depression measurement and composition. In dataset 3, greater reinforcement tendency was specifically associated with a transdiagnostic anxious-depression factor, whereas behavioral persistence was linked to a compulsivity and intrusive thought factor. In this study, depressive psychopathology was associated with a greater tendency to be reinforced by a type of social reward on Twitter, in contrast to typical laboratory-based findings of blunted reinforcement learning in depression. These findings suggest potential mechanisms linking social media use to worse mental health or vice versa, and emphasize the importance of studying real-world behavior.

Hairy root transformation systems are increasingly recognized as effective tools for functional genomics and gene editing studies in various crop species. In this study, we employed the RUBY reporter gene as a visible marker to establish a rapid, simple, and efficient in vivo system for potato hairy root induction and transformation. Explants were wounded by transverse cuts and gently scraped across the surface of a culture plate containing Rhizobium rhizogenes harboring the desired construct, then directly planted into pots containing moist, sterile vermiculite. The 2-3-week-old shoots regenerated from tubers of the Atlantic potato cultivar were identified as sufficient explants for Rhizobium rhizogenes-mediated transformation, achieving transformation efficiencies of up to 100%. This procedure was further adapted for stem nodes and leaf explants and achieved high transformation efficiencies of 80% and 93%, respectively. In addition, the in vivo hairy root transformation system was successfully utilized to validate CRISPR/Cas9 activities by inducing targeted mutations of the StDL1 gene in potato. Heteroduplex analysis on PAGE revealed a high editing frequency (96%), and Sanger sequencing of selected lines confirmed diverse deletions at the target site. Overall, this in vivo hairy root transformation method enables rapid and robust assessment of genome editing efficiency and transgene expression, providing a valuable tool for functional genomics and genetic improvement in potato, with potential applications in other Solanaceae crops.

To explore hypermethylated gene markers in cervical scraped cells that may be associated with endometrial cancer and validate their diagnostic role in endometrial cancer. In an exploratory cohort consisting of 40 paired endometrial tissue and cervical scraping samples, high-performance methylation-targeted genes associated with endometrial cancer were identified. In training and validation sets with 347 and 149 participants, respectively, methylated markers from cervical cytology together with other epidemiological and clinical parameters were assessed to determine their accuracy in detecting endometrial cancer. A decision tree was constructed using methylation markers, bleeding symptoms and endometrial thickness on transvaginal ultrasound (TVS). In this exploratory study, eleven genes highly related to endometrial cancer were evaluated using a methylation array. In the training cohort, the highest AUC values for detecting endometrial cancer were 0.93, 0.91, and 0.89 for hypermethylated CDO1, NEFM, and CELF4, respectively. In the validation set, combining methylated CDO1, CELF4, and NEFM achieved a sensitivity of 94.6% (95% confidence interval 85.1-98.9) and a specificity of 92.8% (89.3-95.5) for detecting endometrial cancer. Integration of the endometrial thickness of TVS slightly improved the diagnostic specificity with very low sensitivity. Cervical cytological DNA methylation assays of CDO1, CELF4 and NEFM provide a reliable and safe strategy for detecting endometrial cancer and are superior to other noninvasive evaluation methods or parameters.

The e-cigarette market evolves faster than regulation and enforcement. In recent years, e-cigarette manufacturers have introduced vapes featuring not just addictive, flavored nicotine, but also technological features that appeal to youth. These products with digital screens have the potential to gamify the use of an addictive drug. To keep pace with these developments, automated tools capable of rapidly analyzing the online marketplace could inform regulatory enforcement. The goal of this work is to automatically detect and classify e-cigarette devices with screens. To do this, we first use an object detection model to filter web-scraped images that contain e-cigarettes. Product images and descriptions are then provided to a vision language model (VLM) to determine if the e-cigarette contains a screen. We evaluated the pipeline with images and text descriptions from five e-cigarette online retailers and manually validated predictions. The object detection model was trained on approximately 7000 images and tested on 3920 images achieving an accuracy, precision, recall, and F1 score of 0.95, 0.98, 0.94, 0.96 respectively in identifying e-cigarette devices. The VLM was tested on 2401 images and was able to detect screens on e-cigarette devices with an accuracy of 0.92, precision of 0.96, recall of 0.87, and F1 score of 0.92. Object detection models and VLMs can be combined to automatically detect and classify e-cigarettes with integrated screens. This method can enable rapid-response pipelines to identify youth-appealing features in emerging e-cigarette products and provide policymakers insights into their proliferation in the online marketplace. New e-cigarettes with potentially youth-appealing technological features, such as integrated screens, continue to be introduced to the US market, challenging timely and effective tobacco product surveillance and regulatory efforts. Digital screens on e-cigarettes offer greater user control and make products that deliver addictive, flavored nicotine resemble entertainment devices, rather than tobacco products. Our automated screen detection pipeline provides a scalable, rapid-response tool for monitoring online e-cigarette retailers in near real-time, quantifying the proliferation of products with specific features. This pipeline could be adapted to accommodate a variety of new features for detection, here we use the presence of screens as a test example. Rapid identification and classification of new tobacco products available online could help inform interventions before the sale and use of such products becomes widespread.

Early adolescence is a time of risk for disrupted or insufficient sleep, and widespread use of electronic media may exacerbate this risk. However, findings linking screen time to sleep in adolescent samples are inconsistent, rely primarily on self-report, and seldom consider sleep variability. We examined associations between 24-hour total screen time scraped from mobile devices and actigraphy-assessed indicators of sleep timing and quality across one week (means, variability, and day-to-day fluctuations) in a community sample of 677 adolescent twins (Mean age = 13.53, SD = 1.13; 50.96% female; 52.88% non-Hispanic White, 21.27% Hispanic/Latino), of whom 389 had actigraphy data and 488 had screen time data. For a subsample of Android users (n=171), hours of social media and mobile gaming were also considered. We conducted person-level associations using path analysis, with standard errors corrected for clustering of twins within families, and day-level analyses using three-level models. After multiple testing correction, greater person-level screen time was associated with later bedtime, and more variable wake time and duration, but not efficiency, latency, catch-up sleep, or social jetlag. More social media use was associated with later bedtime and greater duration variability. Mobile gaming was unrelated to sleep. Day-level screen time was weakly associated with later bedtime that night, with no other significant associations. Associations were mixed and modest, but not reliant on adolescent self-report. Findings do not provide strong evidence that moderate screen use is detrimental to sleep, but future studies should consider individual and contextual moderators and more nuanced objective measures.

In the weeks after California's Eaton-Palisades wildfires, structural ash of burned homes and soils from yards and curbsides were sampled for analysis of Pb, As, and eight metals from 32 burned residences. About six months after fires and after US Army Corps of Engineers' Phase 2 cleanups removed 15 cm from structural ash footprints, 17 properties were resampled within about a meter of original sampling sites. Concentrations of metal-(oid)-s in structural ash and residential soils varied between and within properties with significant spatial and temporal patterns. Lead concentrations were highest in structural ash and soil of yards of select pre-1970s homes, the yard and curbside soils likely having Pb contaminants from before fires. Approximately 28% of structural-ash samples exceeded 200 ppm Pb, the residential soil screening level (SSL) of the US Environmental Protection Agency, and 50% exceeded 80 ppm, California's Pb SSL. After cleanup, <5% of samples from within the scraped foundation areas exceeded 200 ppm Pb and 24% exceeded 80 ppm. Interpreting postcleanup risk from Pb and As depends greatly on the California or federal SSLs. Fires released metals from select older homes to residential soils in concentrations comparable to those found in unburned city soils.

This study examines how mainstream Taiwanese newspapers portrayed sexual minorities from 2010 to 2021, a period marked by rising LGBTQ+ visibility and major legal change. Drawing on institutional mediation and legitimacy perspectives, we test three hypotheses: first, that coverage becomes more negative as LGBTQ+ issues gain visibility over time; second, that gay men receive more negative coverage than lesbian women; and third, that major legal and social milestones shift coverage toward more neutral portrayals and a broader range of topics. To evaluate these expectations, we analyze 18,558 articles from China Times, Liberty Times, and United Daily News using web-scraped data, AI (i.e. GPT-4 model) assisted classification of articles as gay with time series regression models. The findings reveal an overall increase in positive news coverage over the decade, but disparities in the frequency and sentiment of coverage between gay and lesbian subjects persist. The study concludes that while there has been progress toward inclusiveness, significant events and milestones have only partially influenced the portrayal of the LGBTQ+ community. The results highlight the ongoing need for efforts toward equitable media representation.

Antidepressant use and withdrawal are often accompanied by side effects such as dizziness, weight gain, and sexual dysfunction. Antidepressants and their associated side effects are stigmatized topics. Social media platforms such as Reddit are considered "safe spaces" by users because they can freely share their experiences and receive support. This pilot study analyzed discussions from the subreddit r/depression to examine how users discuss antidepressant side effects, withdrawal symptoms, and related experiences of depression. We scraped 10 high-engagement threads from the subreddit r/depression using the Python wrapper for the Reddit application programming interface and conducted a 2-step analysis. First, a pilot test was performed using sertraline (Zoloft) threads, followed by an analysis of all antidepressant-related threads. A subset of the data was hand-coded to create and validate regular expressions, which were then used to automatically code the remaining dataset. The resulting coded data were analyzed using epistemic network analysis and complemented with qualitative analysis and elements of semantic networks and hypergraphs. We found that posts were more likely to discuss emotional flattening, sleep, and memory or cognitive issues (Mann-Whitney U=33,235.5; P=.003). Additionally, references to dizziness tended to co-occur with discussions of withdrawal and offers of empathy, while reports of dream-related side effects and requests for personal experiences also co-occurred frequently. By incorporating elements of semantic networks and hypergraphs, we deduced that offers of empathy occurred when users said they experienced dizziness caused by withdrawal, while mentions of "brain zaps" associated with withdrawal often co-occurred with offers of teaching support. Study findings highlight how individuals experiencing antidepressant side effects and withdrawal symptoms use online forums such as Reddit to seek validation, share coping strategies, and provide emotional support to others. The nuanced discussions observed, particularly those related to empathy, symptom management, and shared learning, underscore the role of peer-to-peer networks in normalizing stigmatized experiences and mitigating isolation associated with antidepressant use. Clinicians and digital health practitioners can leverage these insights to better understand patient language, emotional framing, and informational needs outside clinical settings.

Vertical seawalls are extensive and rapidly expanding in coastal cities and ports but provide inadequate habitat for intertidal fauna and flora as they lack necessary refugia from heat and desiccation stresses, lowering intertidal biodiversity. Ecological engineering through retrofitting suitable habitats aims to promote marine biodiversity through habitat provision. Two eco-engineered panel designs with augmented habitat complexity were tested on a seawall in a highly developed subtropical marina. One panel was commercially available, with a repetitive pattern of short extrusions - the Imported Commercially Available Design (ICAD). The other was a locally designed, double-sided panel with deep intrusions and shade behind the panel - the Hong Kong Design (HKD). Taxonomic richness of the intertidal fauna and flora on the panels were compared to flat, concrete panels (flat panel, representing a worst-case scenario) and scraped seawall plots (seawall control) for 18 months. Marine biodiversity on both eco-engineered panels was significantly higher than the flat concrete panels, and the HKD was colonised by the highest taxonomic richness (64 total taxa). The HKD had greater abundance of suspension feeders, higher richness within key functional groups, and enabled colonisation at higher tidal levels compared to the flat panel and seawall control. When compared to nearby vertical seawalls, the HKD and ICAD contributed to an increased within-site &#x3b2; diversity and functional diversity. These results imply that implementing similar eco-engineering of grey infrastructure while reducing other anthropogenic stressors can contribute to the aim of creating biodiverse intertidal ecosystems within coastal cities and ports.

Fungal keratitis is a vision-threatening infection. Keratitis caused by dematiaceous fungi belonging to or related to the order Pleosporales is rare and diagnostically challenging because culture growth may be slow, morphology may be nonsporulating, and species-level identification may require molecular methods. A 44-year-old male forestry worker developed right ocular pain and a corneal ulcer after ocular trauma with a wooden fragment. He was referred to our hospital approximately one month after the injury; this first visit was defined as Day 0 (202X/11/27). At presentation, best-corrected visual acuity was 1.0 in the right eye. Slit-lamp examination showed a localized gray-white stromal infiltrate at the temporal peripheral cornea in the 9-10 o'clock position, with irregular margins and an overlying fluorescein-positive epithelial defect, but without hypopyon. The ocular media were sufficiently clear for detailed fundus examination, and dilated fundus examination showed no clinical evidence of posterior segment involvement or endophthalmitis. B-scan ultrasonography was therefore not performed. Direct microscopy using 10% potassium hydroxide wet mount and Fungiflora Y staining, corneal culture, and a 12-strip/24-target multiplex real-time polymerase chain reaction panel were all negative at Day 0. After temporary cessation of antifungal therapy because of severe drug-induced hepatotoxicity during systemic voriconazole and acetazolamide therapy (aspartate aminotransferase 540 U/L, alanine aminotransferase 1041 U/L), the lesion enlarged. Repeat scraping culture yielded a filamentous fungus on Day 58. The isolate formed black, slightly velvety colonies and dark hyphae, but did not produce diagnostic conidia or spores under the conditions used. Sequencing of the ribosomal DNA internal transcribed spacer region by Sanger sequencing, followed by BLAST comparison, suggested that the isolate belonged to the order Pleosporales, although species-level identification was not achieved. Treatment consisted of stepwise combination therapy, including topical voriconazole, amphotericin B, and natamycin, initial systemic voriconazole, subsequent topical amphotericin B, systemic and topical micafungin, and oral terbinafine. After active infection had clinically stabilized, 20% autologous serum eye drops were added to support epithelial healing. At the final follow-up on Day 268, best-corrected visual acuity was 1.2, with a quiet anterior segment and a stable corneal scar. This case highlights the importance of repeated microbiological sampling and molecular identification in traumatic fungal keratitis when initial tests are negative. It also illustrates the need for cautious, individualized treatment when azole therapy is interrupted because of systemic toxicity. Because species-level identification and antifungal susceptibility testing were not achieved, the therapeutic implications of this single case should be interpreted conservatively.

To characterise the bacterial and fungal spectrum of infectious keratitis (IK) in southern China and to evaluate changes in the bacterial profiles and antimicrobial resistance (AMR) over an 8-year period (2017-2024). This retrospective study included patients with culture-positive IK treated between 2017 and 2024. Corneal scrapings were obtained for microbiological culture and pathogen identification. Antimicrobial susceptibility testing was performed for all bacterial isolates. Microbial distribution and in vitro antibiotic susceptibility were analysed. A total of 2785 microbial isolates were recovered from 2741 patients. Overall, fungal isolates predominated (59.6%), exhibiting a distinct seasonal distribution, with Fusarium (40.2%) and Aspergillus (14.3%) being the most common genera. Among bacterial isolates, Gram-positive organisms were predominant (63.6%). The most frequently identified Gram-positive organisms were coagulase-negative staphylococci (CNS; 34.9%), while Pseudomonas (18.9%) was the most common Gram-negative pathogen. Over the study period, an increase in the proportions of CNS (p < 0.001) and Serratia was observed (p = 0.017), alongside a decline in Pseudomonas (p = 0.009) and Kocuria (p < 0.001). Resistance among Gram-positive isolates increased for penicillin (from 62.3% to 74.4%; p = 0.002) and levofloxacin (from 26.4% to 46.9%; p < 0.001), whereas Gram-negative resistance generally declined. IK in southern China is characterised by persistent fungal predominance and evolving bacterial composition. AMR patterns differ between Gram-positive and Gram-negative organisms, reflecting both shifts in pathogen distribution and species-specific resistance changes, highlighting the importance of continued regional surveillance to guide empirical therapy.

We report the case of a 10-year-old boy with a unilateral periocular rash, initially misdiagnosed and treated unsuccessfully with multiple courses of topical steroids, oral antibiotics, and eye drops. The rash improved temporarily with tacrolimus ointment, but periocular lesions persisted and spread. Although skin scraping was unsuccessful, the patient was diagnosed clinically with tinea blepharociliaris and treated empirically with a 2-week course of oral antifungal, resulting in rapid clinical improvement. The case highlights key clinical signs to avoid diagnostic delays, unnecessary treatments, and patient distress and supports empiric antifungal therapy when diagnostic tests are inconclusive.

Wild canids serve as definitive hosts for numerous Sarcocystis species infecting livestock and wildlife. However, species-level identification in free-ranging carnivores remains difficult, impeded by the morphological resemblance among the sporocysts of various parasite species. Intestinal mucosal scrapings from 40 wild canids: red fox (Vulpes vulpes, n&#x2009;=&#x2009;8), gray fox (Urocyon cinereoargenteus, n&#x2009;=&#x2009;6) and coyote (Canis latrans, n&#x2009;=&#x2009;26) in Pennsylvania, USA, sampled in 2024, were tested for Sarcocystis sporocysts. Sporocysts were detected microscopically in intestinal homogenates digested in Chlorox in 3/6 (50%) gray foxes, 3/8 (37.5%) red foxes and 18/26 (69.2%) coyotes. PCR amplification was successful on 8/18 (44.4%) coyotes, 0/6&#xa0;Gy foxes and 2/3 (66.6%) red foxes. Multi-locus genotyping was performed for the 18S rRNA, 28S rRNA, COI and ITS1 genetic markers, supporting identification of multiple species of Sarcocystis, including Sarcocystis albifronsi-like, S. cruzi, S. gjerdei-like, S. cristata/S. wenzeli-like and an additional undescribed, ungulate-associated Sarcocystis spp. Phylogenetic analysis confirmed clustering with reference sequences with strong support. This study demonstrates that wild foxes and coyotes serve as definitive hosts for multiple species of Sarcocystis, including Sarcocystis cruzi, which uses cattle as intermediate hosts and imposes significant economic burdens on cattle production. Moreover, these findings demonstrate that wild canids harbor diverse Sarcocystis spp., supporting their role in environmental dissemination and potential transmission at the wildlife-livestock interface.

Fungal keratitis is a serious ocular infection that can result in permanent vision loss if not promptly diagnosed and treated. A healthy woman in her early 20s developed keratitis nearly 2&#x2009;weeks after an ocular trauma with an organic material, a cardboard paper, on her left eye. Ocular examination revealed a mid-stromal corneal infiltrate with brush-like borders and hypopyon. The Giemsa stain of the corneal scraping demonstrated branching septated hyphae, and fungal culture grew Trichomonascus ciferrii resistant to amphotericin B. She was successfully treated with topical natamycin 5% drops for 2 weeks, leading to complete eradication of the corneal infection with a residual stromal scar. This case documented T. ciferrii, a rare fungal aetiology in post-traumatic keratitis in a tropical country, successfully treated with natamycin 5%. Despite the virulence of T. ciferrii, the infection was promptly treated, resulting in a good functional vision.

Acanthamoeba keratitis (AK) is a sight-threatening corneal infection that may initially lack typical epithelial or perineural findings and mimic other infectious or inflammatory anterior segment disorders. Endotheliitis-like presentations of AK with stromal edema, keratic precipitates, Descemet membrane folds, and anterior chamber inflammation have been reported, but progression to severe stromal necrosis with spontaneous sloughing of necrotic corneal tissue is rarely documented. A 48-year-old contact lens wearer presented with right eye pain, redness, and decreased vision. Initial examination at a local clinic revealed anterior chamber inflammation without epithelial defects, radial keratoneuritis, or a ring infiltrate, and topical betamethasone was started before referral. At referral (0&#x2009;weeks; 12&#x2009;days after symptom onset), slit-lamp examination revealed diffuse stromal edema, fine keratic precipitates, Descemet membrane folds, and anterior chamber inflammation, closely mimicking viral corneal endotheliitis. Aqueous humor polymerase chain reaction for human herpesviruses 1-8 was negative. Because viral endotheliitis or noninfectious anterior uveitis was considered, topical corticosteroids and systemic prednisolone were administered (initially 30&#x2009;mg/day and was gradually tapered). During follow-up, the clinical appearance evolved toward AK, and anti-infective therapy was revised. Corneal scraping was performed, but direct microscopy and bacterial and fungal cultures were negative. PCR testing of a corneal scraping specimen for Acanthamoeba, bacteria, and fungi was negative. Despite treatment, stromal necrosis progressed to severe corneal melting. At 34&#x2009;weeks after referral, necrotic corneal tissue spontaneously sloughed as a sequestrum and was submitted for pathological evaluation. Hematoxylin and eosin and periodic acid-Schiff staining revealed necrotic corneal stromal tissue with inflammatory cell infiltration and multiple double-walled cystic structures consistent with Acanthamoeba cysts, confirming AK. This case illustrates a diagnostically challenging form of AK that initially mimicked corneal endotheliitis, yielded negative scraping-based microbiological tests, and was ultimately confirmed by histopathologic examination of spontaneously sloughed necrotic stromal tissue. Severe AK rarely manifests as necrotic stromal sequestration and spontaneous sloughing, in addition to the more commonly recognized destructive outcomes such as corneal thinning, perforation, or keratoplasty-requiring disease. Repeated diagnostic reassessment is important in contact lens wearers with atypical endotheliitis-like keratitis, particularly when the disease progresses despite antiviral or anti-inflammatory therapy.

We investigated the therapeutic effects of combined erythromycin and budesonide treatment on tracheal stenosis (TS) and the associated changes in macrophage-related markers and HDAC2 expression. A rabbit model of TS was established by mechanical scraping of the tracheal inner wall, and the animals were then treated with erythromycin, budesonide, or their combination. Treatment effects were evaluated through histological analysis. We quantified the mRNA expression of M1 and M2 macrophage markers using quantitative PCR and assessed the protein expression of histone deacetylase 2 (HDAC2) by western blotting. RAW264.7 cells were stimulated with lipopolysaccharide or interleukin-4, and macrophage phenotypic changes after erythromycin and budesonide treatment were assessed by flow cytometry. M1/M2 macrophages were co-cultured with fibroblasts, and alpha smooth muscle actin (&#x3b1;-SMA) expression was detected by immunofluorescence. The rabbit TS model exhibited TS characterized by significant thickening of the tracheal mucosa and submucosa. These histological changes were ameliorated in the treatment groups, with the greatest improvement observed in the combination therapy group. The mRNA expression levels of iNOS, CD206, CD163, and Arg1, as well as the protein expression of HDAC2, were elevated in treated groups. In vitro, erythromycin combined with budesonide altered macrophage phenotypic distribution, with the M2/M1 ratio shifting toward 1.0. Additionally, fibroblast &#x3b1;-SMA expression varied according to macrophage phenotype in the co-culture system. Erythromycin combined with budesonide effectively ameliorated injury-induced TS and was associated with changes in macrophage-related markers and increased HDAC2 expression.

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Columnaris is an important bacterial disease of freshwater fish and is increasingly recognised as being caused by a species complex historically referred to as Flavobacterium columnare. A two-step molecular workflow was applied to ornamental fish with clinical signs of columnaris disease from a multispecies commercial distributor operating a shared recirculating system in S&#xe3;o Paulo, Brazil. Skin and fin lesion scrapings were Gram stained, and lesion-derived tissue DNA was screened by ISR-PCR for the columnaris disease complex. ISR-PCR yielded the expected amplicon in 61/123 samples (49.6%) from 28 ornamental fish species. All ISR-PCR-positive samples were then analysed by multiplex PCR for species assignment. Group-specific amplicons were obtained from 29/61 samples (47.5%): Flavobacterium covae in 27/61 (44.3%) and F. davisii in 2/61 (3.3%); no GG1 or GG4 amplicons were detected. The remaining 32/61 samples (52.5%) were not typeable by multiplex PCR. These findings provide baseline species-level data and support combined molecular surveillance in multispecies ornamental fish systems.