Search Results (1,936)

Alpacas have become increasingly popular as farm and companion animals in many countries and were introduced in Hungary approximately 15 years ago. However, reliable data on their population size and management practices have remained limited. Therefore, this study aimed to provide the first nationwide overview of alpaca keeping in Hungary. An anonymous Google Forms-based questionnaire comprising 55 questions was used to collect data on farm characteristics, herd size, husbandry practices, and animal health management. A total of 53 valid responses were analyzed. The results indicated that approximately 260 alpacas are currently kept in Hungary, with most herds being small and typically consisting of 1–5 animals. Most owners planned to expand their herds, primarily through domestic breeding or purchases; however, only a minority considered alpaca keeping economically profitable. Responses related to quarantine, veterinary access, and disease management suggested limited knowledge of biosecurity and animal health. While most owners reported high levels of trust in veterinarians, many had difficulty accessing specialists with expertise in alpacas. In conclusion, alpaca keeping in Hungary is a growing but still small-scale sector. Improving owner education and strengthening professional support may enhance animal welfare and reduce health risks, thereby supporting the One Health approach. Full article

Mountain orchards in southern China are characterized by fragmented and complex terrain with a wide slope variation range (5~30°), which easily leads to uneven pesticide distribution and pesticide accumulation on gentle slopes. These issues give rise to core technical bottlenecks such as low pesticide utilization rate, poor operational efficiency, and unclear atomization mechanism, hindering the optimization of pesticide application parameters, causing pesticide waste and environmental pollution, and restricting the sustainable development of the mountain fruit industry. To address this problem, this study designed a slope-classified pipeline layout and developed a high-efficiency fixed pipeline system for phytosanitary application in mountain orchards, featuring stable operation, low labor intensity, and easy intelligent transformation. Following the technical route of “theoretical design-atomization mechanism analysis-parameter optimization-laboratory verification-field application”, ruby nozzles with high wear resistance, uniform droplet distribution, and long service life were selected and optimized to meet the demand for long-term fixed pesticide application in mountain orchards. High-speed imaging technology was used to real-time capture the dynamic atomization process of nozzles, providing support for clarifying the atomization mechanism. Advanced methods such as fluorescence tracing were adopted to quantitatively evaluate key indicators including droplet deposition in canopies, and the system performance was verified through laboratory and field tests, laying a scientific foundation for its popularization and application. Field test results showed that the optimal spray pressure should not be less than 8 MPa. The XR9002 nozzle can generate fine droplets to achieve pesticide reduction while forming a stable hollow cone atomization flow. Fluorescence tracing analysis indicated that the droplet deposition on the adaxial leaf surface decreases with increasing altitude (presumably affected by wind speed), while the initial deposition on the abaxial leaf surface is low and shows no significant variation with altitude. Deposition on the adaxial leaf surface decreased with canopy height, while abaxial deposition was much lower (8.9–14.9%). This technology enables high-precision quantitative analysis of droplet deposition. The core innovations of this study are: clarifying the atomization mechanism of ruby high-pressure nozzles under pesticide application conditions in mountain orchards, constructing a slope-classified terrain-adaptive pipeline layout model, and establishing a closed-loop technical system of “atomization mechanism-pipeline layout-parameter optimization-deposition detection”. This study provides theoretical and technical support for green and precision pesticide application in mountain orchards, and has important academic value and broad application prospects for promoting the intelligent upgrading of the fruit industry in southern China. Full article

Mythimna separata, an important insect pest of grain crops, exhibits density-dependent prophylaxis (DDP). Most studies have focused on DDP from the perspective of immune system regulation. In this report, we show that two cuticular protein genes, MsCP1 and MsCP2, are involved in DDP of M. separata. MsCP1 and MsCP2 were highly expressed in 4th and 5th instar larvae of the M. separata gregarious high-density phase as compared to solitary, low-density individuals. The expression levels of MsCP1 and MsCP2 were significantly higher in the cuticle of gregarious larvae than in solitary individuals. When MsCP1 and MsCP2 were knocked down by star polycation (SPc) nanocarrier-mediated RNA interference (RNAi), the expression levels of the two genes were significantly downregulated in gregarious larvae as compared to dsGFP/SPc-treated gregarious larvae but were not significantly different from dsGFP/SPc-treated solitary larvae. Furthermore, silencing of MsCP1 and MsCP2 caused abnormal molting, alterations in ultrastructure, and reduced cuticle thickness in gregarious larvae. The mortality rates in dsMsCP1/SPc- and dsMsCP2/SPc-treated gregarious larvae were significantly higher after infection with the entomopathogenic fungus Beauveria bassiana as compared to the dsGFP/SPc-treated gregarious larva but were not significantly different from mortality rates in dsGFP/SPc-treated solitary larvae. Taken together, these findings suggest that upregulation of MsCP1 and MsCP2 are associated with resistance of gregarious M. separata to B. bassiana. These results broaden our understanding of insect DDP and are of great significance in the biological control of M. separata. Full article

Background: Bovine tuberculosis (bTB) caused by Mycobacterium bovis is a major zoonotic disease in West Africa. In Africa, bTB is endemic in cattle with a prevalence ranging from 2% up to 18%. The disease causes significant public health risks due to unpasteurized milk and milk product consumption. In the context of the EU-PRISMA project, which promotes research and innovation for productive, resilient, and healthy agropastoral systems in West Africa, a cross-sectional survey was conducted in dairy herds from Mali and Niger to assess animal, herd, and within-herd bTB prevalence, as well as to identify animal risk factors and predictors of bTB herd status. Method and principal findings: A random cross-sectional survey on dairy cattle farms using comparative intradermal tuberculin test and epidemiological inquiry was performed in four regions of Mali (Bamako, Koulikoro, Mopti, and Sikasso) and three regions of Niger (Tahoua, Dosso, and Tillabéry). Herd and animal prevalence of bTB and within-herd prevalence were significantly higher in Mali (especially in Bamako and Koulikoro) than in Niger. Several risk factors were significantly associated with animals positive to bTB, i.e., the region where animals live, the age range from 3 to 7 years old, and female animals. In addition, in regions with higher bTB prevalence, the herd with slaughtering of animals in the farm and the herd with the presence of an animal assembly area were associated with the most unfavorable status of a herd with regards to bTB. Moreover, the average and the median annual economic losses of bTB at animal level were estimated at €262 and €137 respectively, with large variability depending on the farm (between €46 and €838). Conclusion and significance: This survey provides useful data on bTB epidemiology and economical losses in Mali and Niger and urges for improvement of surveillance systems and prevention and control strategies. Cost-benefit, return of investment, or similar analyses are strongly recommended to help with decision making. Full article

Lactococcosis caused by Lactococcus garvieae is an emerging threat to warmwater aquaculture, yet evidence integrating field outbreaks with robust molecular confirmation and controlled virulence testing remains limited for Thailand’s cage-cultured tilapia. From May to October 2025, acute mortality events were investigated in cage-cultured Nile tilapia (Oreochromis niloticus) in a reservoir in Ubon Ratchathani Province, Thailand. Suspected outbreaks were defined by abrupt daily mortality exceeding 5% accompanied by septicemia-like clinical signs. Water quality during sampling covered the following ranges: temperature 28.6–31.9 °C, pH 6.5–7.0, salinity 0.02–0.03 ppt, electrical conductivity 0.036–0.046 mS/cm, TDS 22.20–26.50 mg/L, total alkalinity 17.0–34.0 mg/L as CaCO₃, total hardness 12.0–60.0 mg/L as CaCO₃, dissolved oxygen 6.5–7.0 mg/L, and NH₃ were below the limit of detection. Full-length 16S rRNA tissue profiling revealed strong tissue partitioning: blood microbiomes were consistently dominated by Lactococcus and L. garvieae at the species level, whereas gills showed higher richness and mixed communities with multiple opportunistic taxa. Culture isolation was more reliable from blood than gills, yielding 16 Gram-positive, catalase-negative isolates (AAHM-LG2501–AAHM-LG2516) that clustered within the L. garvieae clade in near full-length 16S rRNA phylogenetic analysis and were separated from closely related Lactococcus lineages. A representative blood isolate (AAHM-LG2501) showed dose-dependent virulence in controlled challenges, with an LD₅₀ of ~1.05 × 10⁵ CFU/fish by intraperitoneal injection and an LC₅₀ of ~1.20 × 10⁶ CFU/mL by immersion. Histopathology supported systemic dissemination, with injection producing more consistent multi-organ lesions than immersion, particularly in head kidney, liver, and spleen, while gills exhibited route-associated epithelial and vascular alterations. Together, these findings confirm L. garvieae as a major etiological agent of septicemic outbreaks in cage-cultured tilapia in Thailand and support a practical surveillance framework prioritizing blood sampling, molecular confirmation, and risk-based monitoring to guide biosecurity and vaccine-oriented prevention. Full article

Background: Parents serve as the primary decision-makers for childhood vaccination while also making decisions regarding their own vaccination, yet vaccination decision drivers are typically studied separately by vaccine type or target population. Methods: This study investigated parental decision-making processes for two self-paid and non-National Immunization Program vaccines in China, childhood rotavirus vaccine and adult influenza vaccine, by utilizing a structured survey grounded in the World Health Organization Behavioral and Social Drivers of Vaccination framework. Spearman’s rank correlation coefficients were used to assess the consistency of parental attitudes toward the two vaccines across behavioral and social driver domains. Structural equation models were conducted separately for childhood and adult vaccines to examine decision-making pathways. Results: The findings indicated that parental drivers related to awareness, social processes, and practical issues showed a high consistency across adult and childhood vaccination decisions (r > 0.7), whereas the consistency in vaccination behaviors remained low (r = 0.21). Compared with adult vaccination, childhood vaccination decisions were more strongly influenced by vaccine safety concerns and healthcare practitioners’ recommendations, which emerged as key drivers. Furthermore, family norms emerged as an effectively shared driver of vaccination decisions for both adult and childhood vaccines (adult: β = 0.784; childhood: β = 0.970). Conclusions: By jointly synthesizing adult and childhood vaccination decisions from a parental perspective, this study provides crucial evidence to support the development of integrated, family-centered strategies to improve vaccine uptake. Full article

Cryptosporidium spp. are common protist pathogens, and the growing popularity of pet rodents raises concerns about their potential role in zoonotic parasites transmission. However, epidemiological data on Cryptosporidium spp. in pet rodents in Yunnan Province is scarce. To examine the occurrence of Cryptosporidium spp. in pet rodents in Yunnan, we collected 762 fecal samples from four rodent species across four cities. Nested PCR and DNA sequencing were used to characterize the species and subtypes of Cryptosporidium. The occurrence of Cryptosporidium spp. in guinea pigs (Cavia porcellus), Siberian dwarf hamsters (Phodopus sungorus), Syrian hamsters (Mesocricetus auratus) and fancy rats (Rattus norvegicus domestica) was 18.7% (80/426), 17.3% (36/207), 12.5% (15/120), 0% (0/9), respectively, with an overall rate of 17.2% (131/762). According to regions, the positivity rate of Cryptosporidium spp. in Zhaotong city, Kunming city, Yuxi city and Qujing city was 21.0%, 17.9%, 16.8% and 10.5%, respectively. In terms of sampling location, the occurrence of Cryptosporidium spp. in pet markets, farms and shops was 19.5%, 18.6% and 0%, respectively. Sequence analysis of the small subunit ribosomal RNA (SSU rRNA) gene identified six Cryptosporidium species/genotypes: Cryptosporidium homai (n = 52), Cryptosporidium wrairi (n = 30), Cryptosporidium sp. hamster genotype (n = 25), Cryptosporidium andersoni (n = 20), Cryptosporidium parvum (n = 5), and Cryptosporidium muris (n = 1). Further subtyping of C. andersoni isolates using multilocus sequence typing (MLST) revealed a single subtype, with all isolates identified as A3A4A2A2. All five C. parvum isolates were identified as subtype IIdA15G1 based on the gp60 gene. Our findings demonstrated the presence of the zoonotic C. parvum IIdA15G1 subtype in pet rodents, suggesting that these animals, particularly hamsters, may serve as reservoirs for human-pathogenic Cryptosporidium species. These results underscore the need for improved biosecurity and husbandry practices in the pet rodent trade to mitigate public health risks. Full article

International timber trade has accelerated the global spread of the invasive red-haired pine bark beetle H. ligniperda, posing persistent challenges to phytosanitary inspection and border biosecurity. Rapid isothermal amplification assays are increasingly deployed in frontline quarantine settings to support timely regulatory decisions. However, their performance under the heterogeneous biological backgrounds typical of traded timber remains insufficiently evaluated, particularly with respect to the practical implications of low-level false-positive signals. We re-evaluated a previously reported isothermal assay for H. ligniperda using conditions that simulate timber transport and routine customs workflows. Fifty non-target arthropod species (predominantly insects), selected from quarantine interception records, were included to represent taxa likely to co-occur in operational contexts. Material from Lema decempunctata consistently generated weak but reproducible amplification signals across replicates. Sanger sequencing excluded contamination, confirming low-level non-target amplification in complex biological matrices. Although the signals were faint, ambiguous results in quarantine settings may trigger shipment detention, confirmatory laboratory testing, or temporary trade restrictions, thereby increasing inspection workload, delaying clearance, and generating avoidable compliance costs. These findings indicate that trade-mediated species assemblages can compromise assay performance beyond expectations derived from conventional taxonomy-based specificity testing. To reduce interpretive uncertainty and associated regulatory burden, we propose a tiered diagnostic workflow combining rapid on-site isothermal screening with specificity-oriented SYBR Green qPCR confirmation. This strategy enhances diagnostic reliability while preserving operational efficiency in applied biosecurity surveillance. Full article

Glaesserella parasuis (formerly Haemophilus parasuis, HPS), Actinobacillus pleuropneumoniae (APP), Streptococcus suis (SS), and Pasteurella multocida (PM) are common bacterial pathogens associated with Porcine Respiratory Disease Complex (PRDC), a major cause of economic losses in the swine industry. To address this, a cross-sectional study was conducted across 27 large-scale swine farms in Xinjiang, China (October 2024–May 2025). A total of 1239 clinical samples were analyzed by species-specific PCR, and positive samples were further serotyped. Overall, SS and HPS were the predominant pathogens, with higher detection rates in winter and spring. Notably, SS and HPS were most frequent in nasal swabs, while APP and PM predominated in tissue samples. Furthermore, co-infections were common, with HPS + SS being the most prevalent. Serotyping revealed dominance of HPS serotype 12, APP serotype 12, SS serotype 3, and PM serotypes A and B (serotypes E and F not detected). In addition, SS was also detected in environmental samples and farm workers’ nasal swabs. These findings suggest that future prevention and control strategies should focus on developing multivalent vaccines targeting the predominant serotypes identified, implementing regular serotype surveillance to guide precision immunization protocols, and strengthening environmental disinfection and biosecurity practices to reduce co-infections and occupational exposure risks. Full article

Objectives: Platinum resistance remains a critical bottleneck in ovarian cancer management, yet reliable pre-treatment predictive tools are lacking. Existing markers like the platinum-free interval are retrospective, while genomic profiling is often cost-prohibitive. This study aimed to develop an accessible, machine learning-based dynamic weighted fusion (DWF) model using routine laboratory data to provide bidirectional risk stratification, particularly to reliably rule out platinum resistance before treatment initiation. Methods: In this retrospective study (2019–2023), seventy baseline clinical features were collected to differentiate platinum-resistant from platinum-sensitive ovarian cancer patients. We developed a DWF framework that dynamically integrates the top-performing classifiers from a library of 168 algorithms (combining 14 feature selection and 12 machine learning methods). Class imbalance was addressed via oversampling, and model efficacy was evaluated using area under the curve (AUC), accuracy, sensitivity, and specificity. Results: The DWF model achieved a robust AUC of 0.760 (95% CI: 0.683–0.837), outperforming all individual base classifiers. Subgroup analysis demonstrated highly consistent overall discrimination across initial treatment strategies (AUC of 0.755 for primary debulking surgery and 0.761 for neoadjuvant chemotherapy). Feature interpretation highlighted that resistance is driven by synergistic dysregulation of systemic inflammation and hypercoagulability, rather than single biomarkers. Conclusions: The proposed DWF model effectively leverages low-cost, standardized clinical data to serve as a robust bidirectional stratification tool. Its exceptional ability to rule out resistance provides clinicians with the evidence-based confidence to proceed with standard therapies, while its high-risk alerts identify candidates for early therapeutic adjustments and enhanced surveillance in ovarian cancer care. Full article

Tomato (Solanum lycopersicum) is a widely consumed vegetable crop and an established model system for plant functional genomics and genetic research in dicotyledons. Salt stress is a major abiotic factor limiting tomato productivity worldwide. The WRKY transcription factor family, one of the largest and most conserved plant-specific transcription factor families, plays pivotal roles in stress responses. This review summarizes recent advances in understanding the functions of tomato WRKY genes under salt stress, focusing on the genomic basis and evolutionary characteristics of the WRKY family, the roles of core WRKY members under salt stress, and the multi-layered regulatory networks mediating WRKY-dependent salt and alkali tolerance. To date, approximately 10 core SlWRKY genes have been functionally validated to regulate tomato salt tolerance, mainly by maintaining ion homeostasis, regulating reactive oxygen species (ROS) balance, facilitating osmotic adjustment, and integrating hormone signaling pathways. Despite this progress, systemic regulatory hierarchies and epigenetic modulation remain poorly resolved. Furthermore, we discuss how specific WRKY members directly regulate downstream effector genes, such as SlSOS1 and SlNHX4. However, direct experimental evidence for the coordination between tomato WRKYs and mitogen-activated protein kinase (MAPK) cascades, as well as epigenetic modifiers under salt stress, is still scarce in current studies. This review provides a theoretical framework and outlines potential technical pathways for translating fundamental insights into tomato salt tolerance into practical applications for sustainable agriculture. Full article

Wildlife health surveillance is a vital element of disease prevention, biodiversity conservation, and public health protection, especially as most emerging infectious diseases originate from wildlife. In Slovenia, long-term passive surveillance based on necropsy data has yielded valuable insights into wildlife mortality patterns over the past three decades, despite inherent limitations such as carcass detectability, reporting bias, scavenging, and decomposition. Ongoing cooperation among governmental institutions, veterinary services, hunters, and wildlife management organisations has enabled the effective operation of this system, although passive surveillance remains subject to spatial, temporal, and species-specific biases. Necropsy data show that infectious diseases, particularly parasitic infections, are the main causes of mortality in key species such as roe deer and chamois, reflecting both their population abundance and targeted monitoring. In contrast, carcasses of species such as wild boar, red deer, small mammals, and birds are underrepresented due to ecological factors, biosecurity constraints, or low detectability. Overall, while passive wildlife surveillance does not provide representative population-level mortality estimates, it remains a reliable tool for identifying the presence or absence of significant diseases and for understanding broad mortality patterns when interpreted in the context of known methodological and ecological limitations. Full article

Corolla movement is a typical plant movement behavior that enables plants to optimize pollination and adapt to environmental changes. Nevertheless, its molecular mechanism remains poorly understood. In the present study, we conduct a comprehensive transcriptome analysis of Mirabilis jalapa (Nyctaginaceae) corolla at five stages (AG-EG) to elucidate the regulatory networks underlying movement. The results showed that the differentially expressed genes (DEGs) were mainly associated with cellular processes, catalytic activity, MAPK signaling, plant hormone signal transduction, and photosynthesis-related pathways, highlighting their involvement in corolla dynamics. Transcriptome profiling further demonstrated that auxin, ethylene, and abscisic acid signaling pathways were key hormonal regulators of corolla movement. Moreover, Ca²⁺ transport genes (CNGCs and CMLs) and respiratory burst oxidase homologs (RBOHs) were significantly enriched, indicating that Ca²⁺–ROS signaling oscillations also play an important role in driving differential cell expansion and turgor changes. Transcription factor analysis also revealed the upregulation of WRKY2, WRKY22, and WRKY33, suggesting that WRKYs act as the critical transcriptional regulators linking ROS–Ca²⁺ signals with downstream gene expression. The reliability of RNA-Seq data was confirmed by RT-qPCR, which showed high consistency with transcriptome profiles. These findings suggested that corolla movement in M. jalapa is carried through the integration of hormonal pathways, Ca²⁺–ROS signaling, and WRKY-mediated transcriptional regulation. This research provided novel insights into the molecular basis of plant movement and established a foundation for further study on floral dynamics and adaptive strategies in angiosperms. Full article

Background: Rabies is a highly fatal zoonotic disease that causes approximately 59,000 human deaths worldwide each year. Current inactivated rabies vaccines require multiple doses and are associated with high costs. The full-length rabies virus glycoprotein (RVG), a membrane protein, exhibits substantial instability in its trimeric structure during recombinant expression. This instability makes it difficult to obtain high-purity, correctly folded antigens. Objectives: This study focuses on the preparation of a full-length recombinant RVG subunit vaccine candidate expressed in a glycoengineered Pichia pastoris system with mammalian-like glycosylation. Methods: The full-length RVG gene (including the transmembrane domain and cytoplasmic tail) from the Challenge Virus Standard-11 (CVS-11) strain was codon-optimized and inserted into the pPICZαA vector to construct the recombinant expression plasmid pPICZαA-RVG. The plasmid was transformed into glycoengineered Pichia pastoris X33-7 (low-mannose type) by electroporation for inducible expression. The target protein was purified by nickel affinity chromatography, anion-exchange chromatography, and Superdex-200 size-exclusion chromatography. The structural characteristics of the purified protein were analyzed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The purified antigen was formulated with the adjuvants AS03 or MF59. BALB/c mice (n = 5 per group) were immunized intramuscularly following a four-dose schedule (days 0, 7, 14, and 28). Antigen-specific IgG antibody titers were measured by ELISA, and neutralizing antibody titers were determined using the rapid fluorescent focus inhibition test (RFFIT). Results: Glycoengineered Pichia pastoris yeast strains expressing wild-type RVG (RVG-WT) or a mutant variant (RVG-M6: R84S, R199S, H270P, R279S, K300S, and R463S) were successfully constructed. The purified RVG antigen formed nanoparticles with an average particle size of approximately 75 nm. Immunized mice generated robust RVG-specific IgG responses, with titers reaching approximately 6.31 × 10⁵ for RVG-WT after the fourth immunization, compared to 3.16 × 10³ for RVG-M6 and 5.62 × 10³ for the RVG-WT-PEG control. Two weeks after the fourth immunization, RVG-WT formulated with AS03 or MF59 induced significant neutralizing antibody responses compared with the control group (p < 0.0001 and p < 0.01, respectively). The neutralizing antibody titers reached 1:79.43 in the AS03 group and 1:33.11 in the MF59 group, whereas the WT-PEG + AS03 control group showed a low titer of 1:3.72. In contrast, RVG-M6 formulated with MF59 failed to induce detectable neutralizing antibodies (1:3.02). Furthermore, RVG-WT + AS03 induced significantly higher neutralizing antibody responses than the WT-PEG + AS03 control group (p < 0.0001), and a significant difference was also observed between the RVG-WT + MF59 and RVG-M6 + MF59 groups (p < 0.01). Conclusions: The glycoengineered Pichia pastoris expression system successfully produced uniform full-length rabies virus glycoprotein nanoparticles with high purity. When formulated with the AS03 adjuvant, RVG-WT induced high-titer neutralizing antibodies in mice, suggesting a promising strategy for the development of recombinant subunit vaccines against rabies. However, this study is limited by the absence of challenge studies and validation in target animal species, which will be further investigated in future work. Full article

Fusarium oxysporum f. sp. radicis-lycopersici (Forl) is the etiological agent of tomato Fusarium crown and root rot (FCRR), a devastating soil-borne disease that severely compromises global tomato production. The pathogenicity of Forl has been increasingly linked to its capacity to produce phytotoxic isocassadiene-type diterpenoids. In this study, Forl was cultured in rice medium to obtain Forl cultures, which were used for the separation and identification of secondary metabolites. After removing the known metabolites, two new isocassadiene-type diterpenoid compounds, namely fusariumic acids I (1) and J (2), were isolated from the ethyl acetate extract. Their structures were identified using spectroscopic data analyses and quantum chemical calculations. This is the first report of the fusariumic acid analogs containing a hydroxyl group at position C–1 in the molecule. Fusariumic acids I (1) and J (2) exhibited significantly inhibitory activities on the hypocotyl elongation of tomato (Solanum lycopersicum) and sesame (Sesamum indicum) seedlings, as well as on the coleoptile elongation of rice (Oryza sativa var. japonica) seedlings at concentrations from 10 to 100 µg/mL. The discovery of two new phytotoxic isocassadiene-type diterpenoids expanded the diversity of secondary metabolites of Forl. Meanwhile, it provided critical insights into Forl-tomato interactions and the candidate lead compounds for the development of new herbicides as well. Full article