The freeze-thaw process in sperm cryopreservation generates reactive oxygen species (ROS) that impair motility, viability, and fertilizing capacity in fish, including the Javaen barb (Systomus rubripinnis), an endemic Indonesian cyprinid of conservation concern. Cryopreservation underpins gene bank development for aquatic genetic resources and supports biodiversity conservation alongside hatchery-based stock enhancement. This study evaluated five natural antioxidants: basil leaf extract (BLE), noni fruit extract (NFE), moringa leaf extract (MLE), sambiloto leaf extract (SLE), and honey-egg yolk (HEY). Each antioxidant was supplemented into the Kurokura's cryopreservation extender across a graded concentration series (BLE 1-4%, NFE and MLE 0.5-1.5%, SLE 6-9%, and HEY 1-2% with a fixed 5% egg yolk) and compared with an antioxidant-free control. Radical scavenging capacity was measured by DPPH assay (IC50: BLE 134.06, MLE 297.04, NFE 422.86, SLE 3010.46 ppm), and post-thaw motility, duration of motility, viability, abnormality, fertilization rate, and ATP bioluminescence were assessed. The most effective doses tested were doses of 1% for BLE, NFE, and MLE, 1.25% for HEY, and 7% for SLE. BLE at 1% gave the best overall profile (motility 85.00 ± 0.88%; duration 7.14 ± 0.38 min; viability 82.00 ± 1.86%; abnormality 18.26 ± 0.16%), significantly exceeding the control (65.00 ± 2.31%; 4.56 ± 0.33 min; 68.00 ± 2.42%; 23.54 ± 0.28%; p < 0.05), whereas SLE sustained the longest motility. All extracts supported fertilization rates above 82% versus 70.08 ± 3.82% in the control, and 1% BLE preserved post-thaw luminescence equivalent to fresh sperm (165,217 vs. 164,087 RLU), interpreted comparatively. These locally available, plant-derived antioxidants offer a low-cost route to germplasm conservation of Javaen barb, strengthening conservation hatchery and broodstock management, sustainable aquaculture production, and food security for inland fisheries communities.

This study evaluated the dose-dependent effects of Lactiplantibacillus plantarum-fermented noni juice (OLP-FNJ) on growth performance and intestinal physiology in Thai native chickens (Surat Thani strain). A total of 240 birds were randomly allocated to four treatments (0, 2.5, 5.0, and 10.0 mL L[-1] OLP-FNJ administered via drinking water) in a completely randomized design and reared for 16 weeks. Growth performance, serum biochemical indices, digestive enzyme activities, intestinal morphology, tight junction-related and immune-related gene expression, and cecal microbiota composition were evaluated. Dose-response relationships were assessed using polynomial regression analysis. OLP-FNJ supplementation exerted a significant linear dose-response reduction in feed conversion ratio over the entire experimental period (3.11 vs. 2.70; linear p < 0.001) and significantly increased body weight gain during the starter phase (280.06 vs. 307.08 g; linear p < 0.05). The greatest numerical responses were observed at 10.0 mL L[-1] without adverse effects. Performance enhancement was associated with increased digestive enzyme activities, greater villus height-crypt depth ratios, upregulation of tight junction-related genes, and reduced expression of pro-inflammatory markers. Cecal microbial diversity increased in a dose-dependent manner, accompanied by shifts in dominant bacterial phyla and an elevated Firmicutes-to-Bacteroidetes ratio. Serum biochemical parameters remained within physiological ranges, indicating systemic safety. In conclusion, OLP-FNJ enhanced nutrient utilization efficiency in a dose-dependent manner and was associated with coordinated changes in digestive, structural, barrier, immune, and microbial functions. Fermented noni juice shows potential as a functional drinking-water additive to support sustainable production of Thai native chickens.

Understanding toxin resistance in insects is key to appreciating niche adaptations but remains challenging due to its often-polygenic basis. A well-known example is the specialized association of Drosophila sechellia with noni fruit (Morinda citrifolia), which is toxic to other insects, including Drosophila simulans and Drosophila melanogaster. The main noni toxin is octanoic acid (OA), but the mechanisms that determine sensitivity or resistance to OA in different species remain unclear. Here, we experimentally evolved D. simulans with increased OA resistance, identifying multiple loci under selection. Cross-referencing these with a genome-wide, OA resistance CRISPR screen in a D. melanogaster cell line highlighted two proteins: Kraken, a putative detoxification enzyme expressed in digestive and renal tissues, and Alkbh7, a mitochondrial protein linked to fatty acid metabolism. Both genes show elevated expression in D. sechellia and OA-resistant D. simulans. In D. melanogaster, kraken mutants are more OA-sensitive, while Alkbh7 overexpression increased OA resistance. Mutation of these genes in D. sechellia reduced OA tolerance. Our identification of genes contributing to OA resistance in laboratory and natural contexts demonstrates how complementary selection approaches can provide insights into complex mechanisms of toxin susceptibility and adaptation. Such methods could have practical applications in the characterization of natural and artificial insecticides.

β2-containing nicotinic acetylcholine receptors (nAChRs) and dopamine D2 receptors (D2Rs) cooperate to shape striatal dopamine (DA) output, yet the mechanisms by which nicotinic-dopaminergic crosstalk influences D2-like autoreceptor-mediated inhibition of DA release remain unclear. Here we use NiCh8, a covalently linked nicotinic-dopaminergic hybrid, as a molecular probe to test whether single-molecule co-engagement can enhance D2-like autoreceptor-mediated inhibition of DA release. NiCh8 binds native α4β2* and α6β2* nAChRs as well as D2Rs, and behaves as a very low-efficacy partial agonist with antagonistic activity at α4β2 nAChRs. In equilibrium slice/synaptosome assays, NiCh8 elicits a modest dihydro-β-erythroidine-sensitive [[3]H]DA release that is absent in α4/α6 double-knockout (KO) synaptosomes. Strikingly, in superfused striatal synaptosomes NiCh8 potently suppresses basal and nicotine (NIC)-evoked DA outflow at nanomolar concentrations; this inhibition is preserved in β2-KO preparations, abolished by sulpiride, and is not reproduced by the parent pharmacophores (NONI and PAMC) alone or in combination. Structure-guided modeling supports a plausible bitopic binding mode at D2R, in which the dopaminergic fragment is predicted to engage the orthosteric site while the nicotinic fragment may contact a secondary binding region, providing a working structural rationale for the observed inhibitory phenotype. Together, these data identify NiCh8 as a hybrid probe that functionally enhances sulpiride-sensitive D2-like autoreceptor-mediated inhibition of DA release and provides a basis for future studies testing the role of D2R secondary-pocket engagement.

This study investigated the effects of noni fruit flavonoids (NFF) on antioxidant status and anti-inflammatory responses in cashmere goats and explored the underlying mechanisms based on liver transcriptomics and metabolomics. Sixteen male Albas cashmere goats were randomly assigned to two groups: a control (CON) group and a group supplemented with 0.1% NFF. The adaptation and trial periods were 2 and 12 weeks, respectively. The results showed that NFF significantly enhanced antioxidant capacity, with mean increases of 32.9%, 20.7%, and 35.5% for total antioxidant capacity (T-AOC), glutathione peroxidase (GPx), and catalase (CAT) activities, respectively, across serum, liver, spleen, and thymus (p < 0.05). NFF also modulated the inflammatory profile, with a mean increase of 18.0% for interleukin (IL)-10 and mean reductions of 18.6%, 19.4%, and 16.7% for IL-1β, IL-2, and tumor necrosis factor (TNF)-α, respectively, across the serum, liver, and thymus (p < 0.05). Integrative transcriptomic and metabolomic analyses suggested that the AMPK signaling pathway, glutathione metabolism, and TGF-β signaling pathway may be critically involved in mediating these effects. GPX3, GPX8, GCLM, NFKB1, and IL1B were identified as key candidate genes, while adenosine-5'-monophosphate (AMP), cysteinylglycine, and pyroglutamic acid were key differential metabolites. Dietary supplementation with 0.1% NFF enhanced the antioxidant levels and anti-inflammatory response in cashmere goats.

Human papillomavirus (HPV) DNA testing is the recommended primary screening method for cervical cancer prevention. However, implementation in resource-limited and geographically challenging settings is constrained by cost, logistics, and time-sensitive screen-and-treat workflow. Sample pooling may improve testing efficiency, but evidence for its feasibility on point-of-care molecular platforms remains limited. We evaluated the analytical performance and feasibility of sample pooling for high-risk HPV (hrHPV) detection using the STANDARD™ M10 HPV point-of-care assay. Archived self-collected vaginal swab samples from a rural cervical cancer screening programme, Programme SUARA, in Sarawak, Malaysia, were used, including four hrHPV-positive samples spanning a range of cycle threshold (Ct) values and seven hrHPV-negative samples. Pooling strategies of 2-in-1, 4-in-1, and 8-in-1 were assessed using a Dorfman two-stage testing approach. Qualitative agreement, sensitivity, Ct shifts, and concordance with individual testing were analyzed. All hrHPV-positive samples were consistently detected across individual and pooled formats, with no false-negative results observed. Sensitivity remained 100% for all pooling strategies, with perfect agreement between pooled and individual testing (κ = 1.0). Pooling resulted in modest Ct shifts, with mean increases of +1.0, +1.7, and +1.8 cycles for 2-in-1, 4-in-1, and 8-in-1 pools, respectively, without compromising qualitative detection, including samples with low viral loads. Sample pooling for hrHPV DNA testing using the STANDARD™ M10 platform is analytically reliable and operationally feasible. These findings support pooled testing as an evidence-based strategy to improve throughput, reduce resource consumption, and enable same-day screen-and-treat workflows in resource-limited and remote settings.

Livestock remain an understudied component of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ecology in Southeast Asia despite close and frequent contact with humans in smallholder farming systems. This cross-sectional study investigated the seroprevalence of SARS-CoV-2 neutralizing antibodies in livestock in Sarawak, Malaysian Borneo. Archived plasma samples collected between June and September 2024 from 180 livestock animals, including pigs, cattle, buffalo, sheep, and goats, were obtained from the Sarawak Veterinary Diagnostic Service Laboratory and tested using the GenScript cPASS surrogate virus neutralization test (sVNT), a species-independent assay detecting antibodies that block the SARS-CoV-2 spike receptor-binding domain (RBD)-ACE2 interaction. Overall, 19.4% (35/180) of samples were seropositive using the manufacturer-recommended cut-off. Seroprevalence varied significantly by species, with the highest prevalence observed in goats (48.6%), followed by sheep (28.6%), cattle (11.4%), buffalo (8.3%), and pigs (2.6%), and also differed by sampling location but not by age group or sex. Most positive samples exhibited low-level inhibition values near the assay cut-off, suggesting weak or transient neutralizing antibody responses. Multivariable logistic regression identified species as the primary factor independently associated with seropositivity. Collectively, these findings are consistent with sporadic spillback exposure of SARS-CoV-2 from humans into livestock in Sarawak, without evidence of widespread or sustained transmission among livestock populations. Periodic, risk-based surveillance of livestock remains warranted to monitor potential changes in host susceptibility as SARS-CoV-2 continues to evolve.

Noni (Morinda citrifolia L.) is a tropical fruit with significant bioactive potential, yet quality variations between geographical origins remain poorly understood. This study employed UPLC-ESI-MS/MS and GC-MS metabolomics to characterize Samoan (WSM-FNJ) and Hainan, China (HN-FNJ) fermented noni juices. A total of 2047 soluble and 283 volatile metabolites were identified, with 726 and 81 differentially accumulated ones, respectively. While both juices exhibited potent antioxidant capacities, WSM-FNJ exhibited superior antimicrobial activity with inhibition zones of 14.67 ± 0.45 mm and 14.97 ± 0.75 mm against Escherichia coli and Staphylococcus aureus, respectively (significantly higher than those of HN-FNJ, P < 0.01). Furthermore, WSM-FNJ more effectively suppressed pneumonia-related pro-inflammatory genes (IL-6, IL-8) in vitro. These differences may be associated with the higher abundance of specific phenolic acids (e.g., 3-(3-hydroxyphenyl)propionic acid) and iridoids in WSM-FNJ. This work provides preliminary data for the screening of origin-associated biomarkers and the targeted functional development of noni-based products.

Cold plasma (CP) is an innovative processing technology that effectively preserves product quality. This study applies CP to noni juice and evaluates its effects on flavor and bioactive compounds. CP reduced PPO activity of noni juice by 22% and increased POD activity by 60%. Total phenolic content increased from 713.18 to 848.12 μg GAE/mL and total flavonoid content peaked at 554.67 μg RE/mL under CP-treatment at 50 V. In contrast, iridoid content remained stable at approximately 1044.13 μg CAT/mL across all treatments. Metabolomics analysis revealed significant compositional alterations in noni juice, with marked increases observed in various flavonoids, terpenoids, and amino acids. Notably, HS-SPME-GC-MS analysis revealed a reduction in off-flavor fatty acids and a increase in esters such as methyl octanoate, methyl decanoate, etc., leading to improved flavor. Overall, CP significantly improved the flavor characteristics of noni juice and concurrently enhanced various bioactive compounds, thereby strengthening its potential for industrial application.