Background and Aim: Infrared thermography has become an increasingly valuable non-invasive imaging technique for monitoring physiological status and detecting abnormalities in horses. However, its application under tropical environmental conditions remains insufficiently investigated, particularly regarding the influence of ambient conditions on measurement accuracy and agreement with conventional temperature assessment methods. This study aimed to evaluate the agreement among a thermographic camera (TC), infrared thermometer (IR), and rectal temperature (RT), characterize body surface temperature distribution before and after exercise, and determine the effects of environmental conditions on temperature measurements in exercising horses under tropical climate conditions. Materials and Methods: Thirty-one clinically healthy horses performing schooling, training, or lunging exercises were enrolled in this prospective repeated-measures observational study. Temperature measurements were obtained using RT, IR at the medial canthus, and TC across the eye, neck, shoulder, back, trunk, and hindquarters regions before exercise, immediately before and after exercise, and during recovery. Environmental variables, including ambient temperature and relative humidity, were simultaneously recorded. Temperature differences among anatomical regions, exercise stages, and measurement methods were analyzed using linear mixed-effects models. Agreement between methods was assessed using Bland-Altman analysis, whereas environmental influences were evaluated using Kendall’s rank correlation. Results: No significant differences were observed between the left and right body sides or among exercise activities. RT consistently recorded the highest temperatures, followed by IR and TC. Thermographic assessment demonstrated that the eye exhibited the highest surface temperature under resting conditions, whereas the back, trunk, and hindquarters showed the greatest post-exercise temperature increases (p < 0.001). Outdoor measurements produced significantly higher TC temperatures than indoor assessments (p < 0.001). IR demonstrated closer agreement with RT, with a mean difference of 1.02°C, whereas TC underestimated RT by 2.88°C and exhibited wider limits of agreement. Ambient temperature showed a strong positive correlation with TC measurements (r = 0.52, p < 0.001) but had minimal influence on IR measurements, indicating superior environmental stability of IR. Surface temperatures generally returned to baseline during the recovery period. Conclusion: Thermographic imaging provides comprehensive visualization of regional surface heat distribution and effectively identifies exercise-induced thermal changes in horses under tropical conditions. However, its measurements are substantially influenced by environmental temperature and should not be used as a surrogate for core body temperature. In contrast, IR demonstrated greater agreement with RT and superior resistance to environmental variation, making it a practical alternative for rapid estimation of body temperature when rectal measurement is impractical. These findings provide valuable guidance for selecting appropriate temperature monitoring techniques for equine health assessment, exercise monitoring, and welfare management in tropical climates. Keywords: ambient temperature, equine thermography, exercise physiology, heat stress, infrared thermometer, surface temperature, thermographic camera, tropical climate.
Background and Aim: The increasing occurrence of antimicrobial-resistant bacteria in livestock environments highlights the need for sustainable alternatives to conventional antimicrobials. Bacteriophages offer a targeted approach for reducing bacterial contamination while supporting antimicrobial stewardship. This study aimed to develop and evaluate a farm-adapted multicomponent bacteriophage preparation for environmental biocontrol in commercial dairy cattle housing. Materials and Methods: Bacterial strains and bacteriophages were isolated from samples collected at a commercial dairy farm. Selected bacteriophages were characterized for host range, efficiency of plating, multiplicity of infection, and host range expansion through phage adaptation. Purified phage clones were subjected to whole-genome sequencing and bioinformatic analysis. A multicomponent bacteriophage preparation was formulated and lyophilized to facilitate storage and field application. The preparation was subsequently applied to selected environmental surfaces in dairy cattle housing, and bacterial recovery before and after treatment was evaluated using microbiological methods. Results: Phage screening and adaptation enabled the selection of bacteriophages with complementary activity against bacterial isolates recovered from the dairy farm environment. Adaptation expanded the lytic activity of selected phages against previously less susceptible bacterial hosts. Genomic characterization supported the selection of phages for inclusion in the final preparation, while formulation and lyophilization yielded a preparation suitable for field application. Following environmental application under commercial dairy farm conditions, qualitative changes in bacterial recovery were observed on treated surfaces, demonstrating the feasibility of using a farm-adapted bacteriophage preparation for environmental biocontrol. The findings also demonstrated the practical integration of bacterial surveillance, phage isolation, adaptation, genomic characterization, formulation, and on-farm application within a single biocontrol strategy. Conclusion: A farm-adapted multicomponent bacteriophage preparation can provide a practical, targeted approach for environmental bacterial control in commercial dairy cattle housing. The combination of locally isolated phages, host range adaptation, genomic characterization, and a field-applicable lyophilized formulation is a key strength of this approach. These findings provide a foundation for further controlled studies using quantitative microbial assessments to determine efficacy, optimize application protocols, and evaluate the contribution of bacteriophage-based environmental interventions to antimicrobial stewardship and One Health strategies in livestock production. Keywords: antimicrobial resistance, bacteriophage adaptation, bacteriophage biocontrol, dairy cattle, environmental biocontrol, livestock production, One Health, phage therapy.
Background and Aim: Avian pathogenic Escherichia coli (APEC), a major cause of colibacillosis in poultry, possesses diverse virulence-associated genes (VAGs) that contribute to extraintestinal pathogenicity. However, comprehensive information on VAG combinations and co-occurrence patterns among APEC isolates in Vietnam remains limited. This study aimed to characterize VAG prevalence, combination profiles, and co-occurrence patterns in APEC and non-pathogenic E. coli (NPEC) isolates from chickens in northern Vietnam. Materials and Methods: From August 2023 to August 2025, 515 spleen samples were collected from healthy and non-healthy chickens on 35 commercial poultry farms in Hanoi, Hai Phong, Phu Tho, Bac Ninh, and Thai Nguyen. A total of 200 E. coli isolates were recovered and screened by multiplex polymerase chain reaction for 19 VAGs: fimC, eaeA, papC, tsh, sfa_foc, iucD, iutA, irp2, fyuA, iucA, iroN, vat, hlyA, hlyF, astA, ompT, iss, cva/cvi, and colV. Isolates carrying ≥5 of nine selected APEC-associated VAGs were classified as APEC; the remainder were classified as NPEC. We compared gene prevalence, combination profiles, and pairwise co-occurrence using appropriate statistical tests and Jaccard similarity analysis. Results: Among 200 isolates, 121 (60.5%) were classified as APEC and 79 (39.5%) as NPEC. APEC isolates most frequently carried hlyF (100%), fimC and iutA (96.7% each), ompT and iucD (95.9% each), iss (90.1%), irp2 (79.3%), and iroN (65.3%). APEC isolates contained up to 19 VAGs, whereas NPEC isolates contained up to 14. Twenty-three distinct VAG combination profiles were identified among APEC isolates, and 103/121 (85.1%) harbored >8 VAGs. In contrast, NPEC isolates displayed 13 less complex profiles containing 3–6 VAGs. Most VAGs were significantly more prevalent in APEC than in NPEC isolates, while papC, fyuA, and eaeA occurred exclusively in APEC. Co-occurrence analyses demonstrated greater complexity and stronger VAG associations among APEC isolates. Conclusion: APEC isolates from northern Vietnam exhibited extensive VAG diversity and complex co-occurrence patterns distinct from those of NPEC isolates. These findings provide a molecular basis for improved APEC characterization, epidemiological surveillance, and targeted control of poultry colibacillosis. Keywords: avian pathogenic Escherichia coli, chickens, colibacillosis, gene co-occurrence, northern Vietnam, poultry, virulence-associated genes, virulence profiles.
Background and Aim: Tilapia aquaculture systems may act as reservoirs of antimicrobial resistance (AMR) because continuous exposure to antibiotics, disinfectants, and heavy metals can select for resistant bacterial populations. Although Escherichia coli is widely recognized as an indicator organism for monitoring environmental AMR, information on reduced susceptibility to disinfectants and heavy metals in Southeast Asian aquaculture remains limited. Furthermore, epidemiological cut-off values (ECOFFs) for these agents have not been established for E. coli isolated from Thai tilapia farms. This study aimed to characterize the susceptibility of E. coli isolates to disinfectants and heavy metals, propose ECOFFs, and investigate biofilm formation and resistance determinants to improve understanding of AMR ecology in tilapia aquaculture. Materials and Methods: A total of 333 E. coli isolates recovered from fish (n = 251) and cultivation water (n = 82) were investigated. Minimum inhibitory concentrations for five disinfectants and three heavy metals were determined using the agar dilution method, and ECOFFs were estimated using ECOFF Finder software. Biofilm formation was quantified using the tissue culture plate assay. We detected disinfectant- and heavy metal resistance genes by polymerase chain reaction. Principal component analysis was used to classify isolates according to resistance gene profiles, and multivariable logistic regression was performed to identify factors associated with the presence of the class 1 integron gene (int1). Results: A bimodal minimum inhibitory concentration distribution was observed for triclosan, supporting an ECOFF of 0.004 μg/mL; 52.6% of isolates exceeded this value, indicating a non-wild-type population with reduced susceptibility. In contrast, benzalkonium chloride, chlorhexidine (CHX), glutaraldehyde, and nitrilotriacetic acid exhibited unimodal distributions consistent with wild-type susceptibility. Among heavy metals, isolates were most susceptible to silver nitrate (minimum inhibitory concentration (MIC)₅₀ = 16 μg/mL), whereas higher MIC₅₀ values were observed for copper sulfate (CuSo4) and zinc chloride (ZnCl2). Most isolates (87.7%) exhibited weak biofilm formation, while strong biofilm producers were uncommon and predominantly recovered from cultivation water. Chromosomally encoded efflux pump genes, particularly ydgE (97.9%) and mdfA (92.8%), predominated, whereas plasmid-mediated disinfectant resistance genes were infrequent. Principal component analysis identified four distinct resistance gene clusters, and logistic regression demonstrated significant associations between int1 and several antimicrobial and disinfectant resistance determinants. Conclusion: This study proposes the first ECOFFs for triclosan, selected disinfectants, and heavy metals in E. coli isolated from Thai tilapia farms and demonstrates the emergence of non-wild-type isolates with reduced triclosan susceptibility. The predominance of chromosomally encoded efflux pump genes and the observed resistance gene clustering suggest that intrinsic resistance mechanisms contribute substantially to bacterial persistence in aquaculture environments. These findings provide baseline evidence to support routine disinfectant susceptibility monitoring, prudent biocide use, and the integration of aquaculture into One Health AMR surveillance programs.
Background and Aim: Improving reproductive efficiency is a major objective in sheep production because increased lamb output directly enhances flock productivity and economic returns. Ovariocytotoxic serum (OCS), an organ-specific hyperimmune preparation produced against ovarian tissue antigens, has been proposed as a biological alternative to conventional hormonal reproductive management. However, its effects on innate humoral immunity and reproductive performance in sheep remain insufficiently characterized. This study evaluated the effects of OCS administration on hematological and humoral immune parameters, estrus and insemination dynamics, and reproductive outcomes in Kazakh Merino ewes under field conditions. Materials and Methods: A controlled, non-randomized field study was conducted during the breeding season using 200 clinically healthy Kazakh Merino ewes allocated into an OCS-treated group (n = 100) and an untreated control group (n = 100). Treated ewes received two subcutaneous OCS injections before the planned insemination campaign. Hematological indices and serum lysozyme, bactericidal, and complement activities were evaluated in a laboratory subset of 20 ewes per group at five sampling points. Estrus detection, artificial insemination, lambing performance, twinning rate, lamb output, and lamb survival were recorded. Reproductive outcomes were analyzed using Fisher's exact test or chi-square test, and effect estimates were expressed as relative risk (RR), odds ratio, and 95% confidence interval (CI). Results: OCS administration was associated with descriptive increases in humoral innate immune parameters. Mean lysozyme activity increased from 29.1% to 37.3%, bactericidal activity from 49.4% to 58.8%, and complement activity from 19.8% to 21.6% around estrus. By day 25 of the insemination campaign, 93.0% of OCS-treated ewes had exhibited estrus and been inseminated compared with 46.0% of controls (RR = 2.02; 95% CI: 1.62-2.52; p < 0.0001), reducing the insemination period from 21 to 14 days. Twin lambings were significantly more frequent in the OCS group (32.0% vs. 14.0%; RR = 2.29; 95% CI: 1.30-4.02; p = 0.004), increasing lamb output from 104 to 129 lambs per 100 ewes. Although lambing rate was numerically higher following OCS treatment (97.0% vs. 90.0%), the difference was not statistically significant (p = 0.082). Conclusion: OCS administration was associated with enhanced humoral innate immune responses, accelerated estrus and insemination dynamics, increased twinning rate, and greater lamb production in Kazakh Merino ewes. These findings support the potential of OCS as a hormone-sparing biological approach for improving reproductive efficiency under field conditions. Nevertheless, randomized placebo-controlled multicenter studies with comprehensive endocrine, immunological, and long-term safety evaluations are required before routine application in sheep breeding programs can be recommended.
Background and Aim: Proteomic characterization of semen provides molecular insights into reproductive function beyond conventional semen evaluation. Although the seminal plasma and sperm proteomes of the common bottlenose dolphin (Tursiops truncatus) have been reported, corresponding information for the Indo-Pacific bottlenose dolphin (Tursiops aduncus) is lacking. This study aimed to characterize the protein composition of seminal plasma and sperm in T. aduncus to identify proteins and biological pathways associated with reproductive function. Materials and Methods: Semen samples were collected from two healthy adult captive male T. aduncus exhibiting >70% sperm motility. Equal volumes of semen from both animals were pooled, and seminal plasma and sperm fractions were separated by centrifugation. Proteins were extracted, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and identified using liquid chromatography-tandem mass spectrometry. Protein identification was performed with Proteome Discoverer 2.5 using a 1% false discovery rate. Functional annotation was conducted using Gene Ontology enrichment and Search Tool for the Retrieval of Interacting Genes/Proteins protein-protein interaction analyses. Results: A total of 199 proteins were identified, including 84 proteins in seminal plasma and 119 in sperm, with only four proteins shared between the two fractions. Seminal plasma proteins were predominantly enriched in carbohydrate and lipid catabolism, proteolysis, and regulation of endopeptidase activity, indicating roles in maintaining the extracellular environment, protecting sperm, and supporting post-ejaculatory survival. In contrast, sperm proteins were significantly enriched in oxidative phosphorylation, aerobic respiration, mitochondrial adenosine triphosphate synthesis, respiratory electron transport, spermatogenesis, sperm flagellum organization, and motility. Protein interaction analysis demonstrated a highly interconnected mitochondrial network in sperm, whereas seminal plasma proteins formed a more dispersed regulatory network. These findings indicate distinct functional specialization of the two semen fractions and suggest that oxidative phosphorylation represents the principal energy-producing pathway supporting sperm function in this marine mammal. Conclusion: This study provides the first comprehensive proteomic characterization of seminal plasma and sperm in the Indo-Pacific bottlenose dolphin. The identified proteins and enriched biological pathways advance understanding of cetacean reproductive physiology, reveal molecular adaptations associated with mitochondrial energy metabolism, and establish a valuable resource for developing fertility biomarkers and assisted reproductive technologies to support the conservation and reproductive management of this Near Threatened species.
Background and Aim: Trichinella nativa is a freeze-resistant zoonotic nematode that circulates predominantly in Arctic and subarctic wildlife and represents an important foodborne parasite affecting both animal and human health. Its remarkable ability to survive prolonged freezing conditions distinguishes it from other Trichinella species and contributes to its persistence in wildlife reservoirs and transmission through the consumption of raw or undercooked game meat. This review summarizes current knowledge on the taxonomy, epidemiology, transmission, pathogenesis, clinical manifestations, and diagnostic approaches for T. nativa, with particular emphasis on recent advances in molecular and immunological detection methods. Conventional diagnostic techniques, including artificial digestion, serological assays, and molecular methods, are critically discussed, along with their advantages and limitations, for species-specific identification. Emerging recombinant antigen-based diagnostic approaches are also reviewed for their potential to improve the sensitivity and specificity of early detection. In addition, the review highlights recent epidemiological findings from endemic regions, including Central Asia, and discusses the importance of wildlife surveillance and integrated One Health approaches for disease prevention and control. Current knowledge gaps, particularly regarding standardized species-specific diagnostics, field validation of novel biomarkers, and harmonized surveillance strategies, are identified. Overall, this review provides a comprehensive overview of T. nativa biology, ecology, and diagnosis, while emphasizing future research priorities to improve early detection, epidemiological monitoring, food safety, and the effective management of trichinellosis in endemic regions.
Background and Aim: Mesenchymal stem cell-derived exosomes (MSC-Exos) have emerged as promising cell-free therapeutics because they retain the regenerative and immunomodulatory properties of their parent cells while minimizing the risks associated with cell transplantation. Canine dental pulp stem cell-derived exosomes (cDPSC-Exos) represent an attractive veterinary regenerative platform because dental pulp is readily accessible through minimally invasive procedures. However, their molecular composition and xenogeneic therapeutic potential remain largely unexplored. This study aimed to characterize canine dental pulp stem cells (cDPSCs) and their derived exosomes, define the proteomic profile of cDPSC-Exos, and evaluate their safety and therapeutic potential for post-castration wound-healing in cats. Materials and Methods: cDPSCs were isolated from canine dental pulp and characterized according to mesenchymal stem cell (MSC) criteria using morphology, colony-forming ability, proliferation, trilineage differentiation, and reverse transcription-quantitative polymerase chain reaction analysis of MSC surface and stemness markers. Exosomes were isolated from conditioned medium and characterized by nanoparticle tracking analysis and CD9 expression. High-resolution mass spectrometry was used for proteomic profiling, followed by functional pathway and protein interaction analyses. A randomized controlled pilot study involving six healthy male cats (n = 3/group) compared topical cDPSC-Exos gel (3.3 × 10⁸ particles/g) with Bioplacenton® following castration. Wound-healing was evaluated using macroscopic wound scores, hematological parameters, and laser speckle contrast imaging. Results: The isolated cDPSCs fulfilled the defining characteristics of MSCs, including multilineage differentiation and expression of MSC-associated markers. cDPSC-Exos exhibited typical exosome characteristics, with a mean particle diameter of 111 ± 24 nm. Proteomic analysis identified 312 proteins, including 22 proteins associated with inflammation regulation, angiogenesis, extracellular matrix organization, cell migration, and tissue remodeling. Western blot analysis confirmed the presence of vimentin and β-actin, supporting the proteomic findings. Topical administration of cDPSC-Exos was well tolerated, without adverse reactions, and significantly improved wound redness, swelling, and pain scores during early healing (p < 0.05), while hematological variables and tissue perfusion remained comparable to the positive control. Conclusion: This study provides the first comprehensive proteomic characterization of cDPSC-Exos and the first evidence supporting their xenogeneic application in feline post-castration wound-healing. The findings demonstrate that cDPSC-Exos constitute a safe and promising cell-free regenerative therapy capable of modulating early wound repair through multiple biological pathways. Larger controlled studies are warranted to validate their clinical efficacy and optimize therapeutic protocols.
Background and Aim: Oil palm by-products are abundant lignocellulosic resources with considerable potential as sustainable ruminant feed but are constrained by poor digestibility and limited rumen fermentability. White-rot fungi (WRF) produce ligninolytic enzymes capable of modifying lignocellulosic biomass, whereas fungal consortia may enhance substrate bioconversion through complementary enzymatic activities. This study evaluated the effects of single and consortium WRF on the nutritional quality, in vitro rumen fermentation, digestibility, methane (CH4) production, and rumen microbial populations of mixed oil palm by-products. Materials and Methods: A substrate comprising oil palm fronds, palm kernel cake, and oil palm decanter cake was subjected to 28-day solid-state fermentation using 6% (w/w) inoculum. Five treatments were evaluated: Unfermented control, fermented control without fungal inoculation, Phanerochaete chrysosporium, Trametes versicolor, and their consortium, each with five biological replicates. Nutrient composition was determined after fermentation. In vitro gas production kinetics, rumen fermentation characteristics, CH4 production, digestibility, and metabolizable energy (ME) were evaluated over 72 h using buffered rumen fluid. Rumen microbial populations were quantified by quantitative polymerase chain reaction. Results: Fermentation significantly increased crude protein (CP) by 24.8%–34.8% while decreasing ether extract, hemicellulose, and non-fiber carbohydrates (p < 0.05). Conversely, relative lignin concentration and structural fiber fractions increased, indicating preferential carbohydrate utilization during fungal growth. All fermented treatments reduced cumulative gas production, total volatile fatty acids, CH4 production, digestibility, and ME compared with the unfermented control (p < 0.05). CH4 production declined by 26.6%–33.6%, although this reduction was accompanied by lower fermentation efficiency rather than improved feed utilization. Ruminal pH and ammonia nitrogen remained within acceptable ranges and were unaffected by fungal inoculation. Total bacterial abundance differed among treatments, whereas the populations of Fibrobacter succinogenes and Ruminococcus albus remained unchanged. The consortium treatment did not provide additional benefits over single-fungus fermentations for nutrient utilization or rumen fermentation responses. Conclusion: Bioprocessing mixed oil palm by-products with single or consortium WRF altered substrate composition by increasing CP while reducing CH4 production. However, these changes were accompanied by reduced rumen fermentability, digestibility, and energy availability, indicating that the tested fermentation conditions did not improve the overall feeding value of the mixed substrate. Consortium fermentation offered no clear advantage over single fungal cultures, highlighting the need to optimize fungal species combinations and fermentation conditions before practical application as a biological pretreatment for ruminant feeds.
Background and Aim: Spotted scat (Scatophagus argus) is a euryhaline teleost of increasing importance in aquaculture, ornamental fisheries, and environmental monitoring across the Indo-Pacific region. Hematological evaluation is a valuable tool for assessing fish health, physiological status, and environmental quality; however, species-specific reference data for spotted scat remain scarce. This study aimed to characterize blood cell morphology and morphometry and establish preliminary hematological reference values for wild-caught spotted scat from Southern Thailand to support veterinary diagnostics, future research, and biomonitoring applications. Materials and Methods: Twenty-six wild-caught spotted scats were collected from Tha Sung Canal, Nakhon Si Thammarat Province, Thailand, between June and August 2025. Fish were anesthetized with clove oil before blood collection from the caudal vein into ethylenediaminetetraacetic acid tubes. Wright-stained blood smears were prepared for microscopic evaluation of blood cell morphology and morphometry. Hemoglobin (Hb) concentration was measured using an automated hematology analyzer, whereas packed cell volume (PCV), erythrocyte and leukocyte counts, differential leukocyte counts, thrombocyte counts, and erythrocyte indices were determined using standard manual hematological methods. Descriptive statistics were generated, and leukocyte morphometry was compared using the Kruskal-Wallis test followed by Dunn’s multiple-comparison test. Results: Four leukocyte types were identified: neutrophils, eosinophils, monocytes, and lymphocytes. Neutrophils and monocytes were the largest leukocytes and exhibited similar diameters (p > 0.05), although monocytes possessed a characteristically deeper blue cytoplasm that facilitated differentiation. Eosinophils occurred as small and large forms, whereas lymphocytes represented the predominant circulating leukocyte population. Juvenile male fish had a mean PCV of 0.24 ± 0.07 L/L, Hb concentration of 74.41 ± 20.94 g/L, erythrocyte count of 1.80 ± 0.68 × 10¹²/L, and leukocyte count of 32.47 ± 14.81 × 10⁹/L. Lymphocytes (16.14 ± 7.27 × 10⁹/L) were the predominant leukocyte subtype, followed by monocytes, neutrophils, and eosinophils. Conclusion: This study provides the first comprehensive description of blood cell morphology together with preliminary hematological reference values for wild-caught spotted scat from Southern Thailand. These baseline data enhance diagnostic capacity for piscine hematology, support health assessment of ornamental and cultured fish, and provide a valuable foundation for future studies investigating environmental stress, disease, and the use of spotted scat as a bioindicator of freshwater, brackish, and mangrove ecosystems within the One Health framework.
Background and Aim: Cannabidiol (CBD) is increasingly administered via electronic vaporizers because inhalation enables rapid pulmonary absorption and bypasses first-pass hepatic metabolism. However, the respiratory consequences of repeated exposure to purified CBD vapor remain insufficiently characterized. This study investigated the systemic and pulmonary effects of repeated whole-body inhalation of purified CBD vapor in Sprague–Dawley rats, with emphasis on the temporal progression of pulmonary histopathological changes. Materials and Methods: Thirty 9-week-old male Sprague–Dawley rats were randomly allocated to five groups (n = 6/group): saline-exposed control and CBD vapor exposure for 15, 30, 60, or 90 consecutive days. Approximately 100 mg of purified CBD isolate was loaded into an electronic vaporization device for each exposure session, and individual rats underwent whole-body exposure for 5 min/day under standardized chamber conditions. Clinical status, body weight, and rectal temperature were monitored. Hematological, biochemical, and electrolyte parameters were measured at the end of each exposure period. Lung and extrapulmonary tissues were examined histopathologically. Pulmonary lesions were independently evaluated by two blinded veterinary pathologists using a semi-quantitative grading scale. Results: No mortality, clinically apparent morbidity, abnormal respiratory signs, or significant differences in body weight were observed. Several hematological and biochemical variables differed significantly among groups; however, most post hoc comparisons were nonsignificant, and all values remained within physiological reference intervals. No histopathological abnormalities were detected in the liver, gastrointestinal tract, kidneys, or brain. In contrast, repeated CBD vapor exposure produced progressive pulmonary inflammatory and vascular alterations. Interstitial pneumonia and bronchus-associated lymphoid tissue hyperplasia were evident from 15 days, whereas perivascular eosinophilic cuffing and pulmonary arterial smooth muscle hypertrophy increased with exposure duration. The most severe lesions occurred after 60 and 90 days and included marked interstitial inflammation, dense eosinophilic perivascular infiltrates, hemorrhagic foci, and advanced arterial remodeling. Conclusion: Repeated whole-body inhalation of purified CBD vapor was associated with progressive, localized pulmonary inflammatory and vascular changes despite the absence of clinically meaningful systemic toxicity. These findings support further studies incorporating aerosol characterization, inhaled dosimetry, pulmonary function testing, mechanistic biomarkers, recovery assessment, and clinically relevant vaping formulations.
Background and Aim: Dried fish sold in informal markets is an affordable source of protein but may be vulnerable to microbial contamination because of inadequate hygiene and handling practices. This study aimed to assess microbial contamination levels, identify bacterial isolates using 16S rRNA sequencing, and determine the antimicrobial resistance (AMR) profiles of bacteria recovered from sun-dried, smoked, and salted fish sold in informal markets in Gauteng Province, South Africa. The research addressed limited information on the microbiological quality and AMR of dried fish sold in South African informal markets, particularly the integration of culture-based, molecular, and resistance-profiling approaches in a One Health framework. Materials and Methods: A total of 140 dried fish samples (80 sun-dried, 40 smoked, and 20 salted) were collected from 12 informal markets in Johannesburg and Pretoria using convenience sampling. Total bacterial counts (TBC) were determined on nutrient agar, followed by phenotypic characterization and molecular identification using 16S rRNA gene sequencing. Antimicrobial susceptibility testing was performed using the disk diffusion method against seven antibiotics. Data were analyzed using descriptive statistics, analysis of variance with Bonferroni correction, and chi-square tests, with statistical significance set at p < 0.05. Results: TBC ranged from 0.8 × 10⁷ to 5.34 × 10⁷ colony forming units/g, with a significant difference between sun-dried and salted fish after Bonferroni adjustment (p = 0.012). Sixty-seven bacterial isolates were identified, dominated by Clostridium spp. (10/67, 14.92%), Staphylococcus spp. (8/67, 11.94%), Staphylococcus xylosus (6/67, 8.95%), and Klebsiella pneumoniae (5/67, 7.46%), including Clostridium botulinum (2/67, 2.98%). High resistance was observed to streptomycin (73.1%) and erythromycin (58.2%), and several bacterial isolates exhibited multidrug resistance. No significant differences in bacterial counts were detected among sampling locations (p = 0.457). Conclusion: This study provides critical evidence of multidrug-resistant bacteria, including C. botulinum, in dried fish from informal markets, highlighting serious food safety and public health risks. Findings underscore the need for enhanced hygiene practices, vendor training, and regulatory oversight to mitigate contamination and the spread of AMR in informal food systems.
Background and Aim: Cholesterol (CHO) sequestration from the sperm plasma membrane is a fundamental event during mammalian sperm capacitation, but the molecular mechanisms underlying capacitation-like sperm activation in brachyuran crabs remain unknown. Although Niemann–Pick type C2 (NPC2), a high-affinity CHO-binding protein, has been implicated in sperm maturation in penaeid shrimp, its occurrence and physiological role in brachyuran species have not been investigated. This study characterized the Portunus pelagicus NPC2 (PpNPC2) gene, determined its localization in the male reproductive tract, and evaluated its role in capacitation-like sperm remodeling during reproductive tract transit in CHO-dependent capacitation. Materials and Methods: The PpNPC2 sequence was identified and analyzed using comparative molecular and phylogenetic approaches. Localization of PpNPC2 in the testis and spermatic duct was examined by immunohistochemistry. Recombinant PpNPC2 protein was produced in Escherichia coli and functionally evaluated using isolated testicular sperm. Membrane CHO content was quantified by high-performance thin-layer chromatography and Filipin III fluorescence staining, whereas sperm morphology and protein tyrosine phosphorylation were assessed using light microscopy, immunofluorescence, and Western blotting. Results: PpNPC2 encoded a conserved NPC2 protein that clustered phylogenetically with crustacean NPC2 homologs and was predominantly localized in epithelial cells of the testis and spermatic duct. During sperm transit, membrane CHO decreased by approximately 32%, accompanied by progressive transformation of round testicular sperm into characteristic snowman-like spermatic duct sperm. Recombinant PpNPC2 reproduced these physiological changes by promoting CHO sequestration, inducing time- and concentration-dependent morphological remodeling, and significantly increasing protein tyrosine phosphorylation by up to 2.33-fold (p < 0.01). These findings indicate that PpNPC2 functions as a CHO-sequestering protein that initiates capacitation-like biochemical and structural remodeling during sperm maturation. Conclusion: This study provides the first molecular and functional characterization of NPC2 in a brachyuran crab and demonstrates that NPC2-mediated CHO sequestration is a conserved mechanism driving capacitation-like sperm activation during reproductive tract transit. These findings expand current understanding of reproductive biology in marine crustaceans and provide a molecular foundation for future studies on sperm preservation, assisted reproduction, and crustacean aquaculture.
Background and Aim: Deep-bedded housing systems have been increasingly adopted as alternatives to conventional pig production because they provide manipulable substrates, improve behavioral opportunities, and support circular manure management. However, their effects on animal welfare, production performance, and environmental sustainability remain inconsistent across production stages, bedding materials, and management systems. Furthermore, evidence comparing individual bedding materials is fragmented, particularly for tropical substrates such as rice husk. This systematic review aimed to critically evaluate the effects of deep-bedded pig systems on animal welfare, productivity, environmental sustainability, and economic performance through qualitative evidence synthesis and quantitative meta-analysis, and to develop practical translational frameworks to support veterinary decision-making. Materials and Methods: A systematic review was conducted according to the PRISMA 2020 guidelines using Web of Science, Scopus, and PubMed to identify studies published between January 2000 and March 2026. A total of 790 records were retrieved, of which 221 comparative studies met the eligibility criteria for qualitative synthesis. Pairwise random-effects meta-analyses were performed for average daily gain (ADG), feed conversion ratio (FCR), and mortality when sufficient quantitative data were available. Risk-of-bias was assessed using Risk-of-bias In Non-randomized Studies of Interventions (ROBINS)-I and RoB 2 principles, whereas certainty of evidence was evaluated using the Grading of Recommendations Assessment, Development and Evaluation. The feasibility of network meta-analysis was also examined. Two translational frameworks, the Healthy Pig Production Framework and the Sustainable Deep-Bedding Model, were developed to integrate scientific evidence into veterinary practice. Results: Deep-bedding consistently improved behavioral welfare by increasing rooting, exploratory activity, and resting comfort compared with conventional housing. Meta-analysis demonstrated no overall effect on ADG (mean difference: +9.8 g/day; 95% confidence interval: −58.3 to 77.9; p = 0.78), although subgroup analysis indicated improved growth in nursery-growing pigs and reduced performance in growing-finishing pigs. Evidence for FCR and mortality remained inconclusive because of incomplete reporting and substantial heterogeneity. Environmental outcomes indicated improved manure recycling potential but greater risks of ammonia emissions and greenhouse gas production when bedding moisture and ventilation were poorly managed. Material-specific evidence was insufficient to support a formal network meta-analysis, highlighting a major research gap. Rice husk emerged as a promising bedding material for tropical production systems, although current evidence remains predominantly qualitative. Conclusion: Deep-bedded housing should be regarded as a management-dependent strategy for welfare and sustainability rather than a simple flooring alternative. Although welfare benefits were consistently observed, production and environmental outcomes varied by production stage, substrate type, and management quality. The proposed translational frameworks provide practical tools for veterinary assessment and sustainable pig production while identifying priorities for future substrate-specific comparative research.
Background and Aim: Effective pain management is fundamental to animal welfare and successful perioperative care. Although multimodal analgesia incorporating opioids is considered the standard of care in companion animal practice, opioid availability and clinical use vary considerably among countries because of differences in regulations, access, and professional training. Information regarding opioid prescribing and pain management practices in the Baltic region is lacking. This study aimed to evaluate opioid availability, opioid use patterns, perioperative pain management practices, and pain assessment approaches among veterinarians in Latvia, with particular emphasis on regional disparities, surgical activity, and generational differences in clinical practice. Materials and Methods: A national cross-sectional survey was conducted using a structured online questionnaire comprising 24 multiple-choice items distributed to licensed veterinarians in Latvia between December 2023 and January 2024. The questionnaire collected information on respondent demographics, opioid availability and use, perioperative analgesic practices, pain assessment methods, adverse effects, and factors influencing opioid selection. Descriptive statistics were used to summarize responses. Associations between categorical variables were evaluated using the chi-square test, whereas relationships between professional experience and opioid availability were examined using Spearman's rank correlation and binary logistic regression. Results: A total of 101 veterinarians completed the survey, representing 10.4% of registered Latvian Veterinary Association members. Opioids were used routinely in 80.0% of practices, most commonly in combination with non-steroidal anti-inflammatory drugs as part of multimodal analgesia. Butorphanol and buprenorphine were the most widely available opioids, whereas fentanyl and methadone availability was significantly associated with higher surgical caseloads (p < 0.01). Fentanyl availability was greatest in Rīga and surrounding regions but was absent in Latgale, demonstrating marked regional disparities. Veterinarians with fewer years of professional experience reported significantly greater access to fentanyl and more frequent use of standardized pain assessment tools. Opioid selection generally reflected perceived pain severity, with butorphanol preferred for mild pain, buprenorphine for moderate pain, and fentanyl and methadone for severe pain. Despite widespread opioid adoption and positive attitudes toward analgesia, only one-third of respondents routinely used validated pain scoring systems. Conclusion: This study provides the first national overview of opioid use and perioperative pain management practices among companion animal veterinarians in Latvia and the Baltic region. The findings demonstrate broad adoption of evidence-based multimodal analgesia while identifying important regional inequalities in opioid availability and persistent underuse of standardized pain assessment tools. These results highlight priorities for continuing professional education, wider implementation of validated pain assessment methods, and improved access to essential analgesics to promote consistent, evidence-based pain management in veterinary practice.
Background and Aim: Sudden acquired retinal degeneration syndrome (SARDS) is an important cause of acute, irreversible blindness in dogs and is frequently accompanied by systemic clinical and biochemical abnormalities. Despite numerous studies describing the disease, no clinically applicable prediction tool has been developed to facilitate early recognition before electroretinographic confirmation. This study aimed to identify demographic, hematological, and biochemical factors associated with SARDS and to develop and internally validate a practical clinical risk score for early identification of affected dogs. Materials and Methods: A retrospective case–control study was conducted using medical records of dogs that underwent electroretinography between January 2018 and December 2023 at the Kasetsart University Veterinary Teaching Hospital in Thailand. Fifty-three dogs with electroretinography-confirmed SARDS were compared with 219 breed-frequency-matched cataract patients exhibiting normal electroretinographic responses. Demographic characteristics, complete blood count results, and serum biochemical parameters were analyzed. Variables associated with SARDS in univariable logistic regression (p < 0.20) were entered into multivariable logistic regression using a manual purposeful selection approach. Independent predictors were incorporated into a weighted clinical risk score. Diagnostic performance was evaluated using receiver operating characteristic analysis, sensitivity, specificity, and area under the receiver operating characteristic curve (AuROC). Results: Dogs with SARDS exhibited significantly higher neutrophil counts, alkaline phosphatase (ALP) activity, plasma protein concentration, alanine aminotransferase activity, creatinine concentration, and white blood cell counts than control dogs. Multivariable logistic regression identified elevated neutrophil count (odds ratio [OR] = 8.80; p < 0.001), elevated ALP activity (OR = 6.80; p < 0.001), and increased plasma protein concentration (OR = 3.94; p = 0.004) as independent predictors of SARDS. Based on optimal cutoff values (neutrophils ≥10,525 cells/μL, ALP ≥74 U/L, and plasma protein ≥7.3 g/dL), a weighted clinical risk score ranging from 0 to 5 points was developed. A threshold score of ≥3 demonstrated good discriminative ability, with an AuROC of 0.83 (95% confidence interval: 0.76–0.90), sensitivity of 74.47%, and specificity of 79.79%, outperforming each individual laboratory parameter. Conclusion: This study identified readily available hematological and biochemical variables that are independently associated with SARDS and developed the first weighted clinical risk score for early identification of the disease in dogs. The proposed model demonstrated good diagnostic performance and may assist veterinarians in prioritizing dogs for ophthalmic evaluation and electroretinography. Although external validation in independent populations is warranted, this practical scoring system represents a valuable adjunct for improving early clinical suspicion and diagnostic decision-making in canine SARDS.
Background and Aim: Femoral head and neck ostectomy (FHO) is a widely performed salvage procedure for canine coxofemoral disorders; however, postoperative recovery is frequently limited by reduced weight bearing, persistent lameness, muscle atrophy, and restricted hip mobility. Although underwater treadmill (UWTM) rehabilitation has been used for various orthopedic conditions, objective evidence supporting its effectiveness after FHO remains limited. This study aimed to evaluate the effectiveness of a standardized 10-session UWTM rehabilitation protocol on functional recovery in dogs after FHO, using clinical and objective gait assessment measures. Materials and Methods: A prospective controlled pilot study was conducted using 10 client-owned dogs that underwent unilateral FHO. Dogs were allocated to either a UWTM rehabilitation group (n = 5) or a non-UWTM control group (n = 5). One dog in the UWTM group was withdrawn due to force plate noncompliance, leaving four treatment and five control dogs to complete the 12-week follow-up. Rehabilitation consisted of 10 weekly UWTM sessions initiated approximately 3 weeks after surgery. Clinical evaluations included lameness and pain scores, thigh circumference, and force plate kinetic gait analysis. Peak vertical force (PVF) normalized to body weight (BW) served as the primary objective outcome measure. Assessments were performed at postoperative weeks 4, 8, and 12. Results: Dogs receiving UWTM rehabilitation demonstrated significant improvements in functional recovery over time. Lameness scores decreased significantly between postoperative months 1 and 3 (mean difference = 2.00 points, p < 0.05), while PVF of the operated limb increased significantly by 23.66% of BW between months 1 and 3 (p < 0.05). Interlimb PVF asymmetry also decreased significantly in the UWTM group. In contrast, the control group showed significant improvement only in lameness score by month 3, whereas PVF did not change significantly throughout the study. Pain scores and thigh circumference did not differ significantly between groups over time. No serious treatment-related adverse events were observed during the rehabilitation protocol. Conclusion: A standardized 10-session UWTM rehabilitation protocol may accelerate postoperative functional recovery following FHO by improving limb weight bearing and reducing lameness without treatment-related complications. Although these preliminary findings support UWTM as a promising adjunctive rehabilitation strategy after FHO, the small sample size and pilot design limit generalizability. Larger randomized controlled trials with extended follow-up are warranted to establish optimal rehabilitation protocols and confirm long-term clinical benefits.
Background and Aim: The increasing restrictions on antibiotic growth promoters have intensified the search for safe and effective phytogenic alternatives for poultry production. Oregano (Origanum vulgare L.) essential oil (OEO) possesses antibacterial, antioxidant, anti-inflammatory, and immunomodulatory activities, but its high volatility and poor thermal stability limit practical applications. This study aimed to develop microencapsulated OEO and evaluate its effects on growth performance, carcass characteristics, and meat quality in broiler chickens. Materials and Methods: OEO microcapsules were prepared by complex coacervation using gelatin and gum Arabic as wall materials. Physicochemical characteristics were evaluated through encapsulation efficiency, oil loading capacity, particle size analysis, Fourier-transform infrared spectroscopy, scanning electron microscopy, thermogravimetric analysis, differential scanning calorimetry, and release behavior studies. A feeding trial involving 3,600 one-day-old Arbor Acres broilers was conducted for 42 days. Birds were randomly allocated to four dietary treatments containing 0, 80, 120, or 160 mg/kg microencapsulated OEO. Growth performance, slaughter traits, meat quality parameters, and proximate composition of breast and thigh muscles were determined. Results: The optimized microcapsules exhibited an encapsulation efficiency of 84.2%, a product yield of 82.37%, and a mean particle size of 151 μm. Structural and thermal analyses demonstrated effective encapsulation and enhanced thermal stability with sustained-release properties. Dietary supplementation with microencapsulated OEO improved broiler performance in a dose-dependent manner. Birds receiving 160 mg/kg supplementation exhibited significantly greater final body weight and average daily gain, together with reduced feed-to-gain ratio and mortality rate (p < 0.05). Breast muscle yield increased, whereas abdominal fat percentage decreased significantly (p < 0.05). Supplementation also improved meat quality by increasing muscle pH, reducing drip loss, enhancing redness, and decreasing lightness. Furthermore, breast muscle crude protein content increased, whereas crude fat content in breast and thigh muscles decreased significantly (p < 0.05). Conclusion: Microencapsulation using gelatin and gum Arabic effectively enhanced the stability and controlled-release properties of OEO. Dietary inclusion of 160 mg/kg microencapsulated OEO improved growth performance, carcass characteristics, and meat quality while reducing fat deposition in broilers. These findings indicate that microencapsulated OEO represents a promising phytogenic feed additive and a potential alternative to antibiotic growth promoters in sustainable poultry production.
Background and Aim: Spontaneous intracerebral hemorrhage (sICH) is a devastating subtype of stroke characterized by high mortality, severe neurological disability, and limited effective pharmacological treatment. Secondary brain injury following sICH is largely driven by excessive neuroinflammation, in which activated microglia predominantly adopt a pro-inflammatory (M1-like) phenotype that exacerbates neuronal damage. The voltage-gated potassium channel Kv1.3 is highly expressed in activated microglia and represents a promising therapeutic target for modulating neuroinflammatory responses. This study investigated whether selective Kv1.3 inhibition with 5-(4-phenoxybutoxy) psoralen (PAP-1) could promote immuno-modulatory microglial polarization, attenuate brain injury, and improve neurological recovery in a collagenase-induced mouse model of sICH. Materials and Methods: Adult male ICR mice with collagenase-induced sICH received daily intraperitoneal administration of PAP-1 (40 mg/kg) or vehicle for up to 7 days. Survival, body weight, hematoma volume, neuronal survival, Kv1.3 expression, and microglial polarization were evaluated using histological and immunohistochemical analyses. Neurological recovery was assessed using the modified neurological severity score, cylinder test, corner turn test, and open field test. Histological analyses were performed on post-injury days 1, 2, 3, and 7, whereas behavioral assessments and survival monitoring were conducted longitudinally throughout the experimental period. Results: Selective Kv1.3 inhibition significantly improved neurological recovery following sICH. PAP-1 treatment increased 7-day survival from 40% to 70% and significantly reduced hematoma volume from day 2 onward, with reductions of approximately 19% on days 2 and 3 and 24% on day 7 compared with vehicle-treated mice. PAP-1 preserved perihematomal neurons, producing an approximately 2.9-fold increase in NeuN-positive neuronal density on day 7. Furthermore, PAP-1 suppressed Kv1.3 expression, reduced CD16/32-positive M1-like microglia, enhanced CD206-positive M2-like microglial polarization beginning on day 1, increased the M2/M1 ratio, and significantly improved sensorimotor and locomotor performance across all behavioral assessments. These findings demonstrate that selective Kv1.3 inhibition effectively attenuated neuroinflammation, accelerated hematoma resolution, and promoted functional recovery after experimental sICH. Conclusion: Selective inhibition of Kv1.3 with PAP-1 promoted a favorable shift toward an immunomodulatory microglial phenotype, reduced hematoma burden, preserved perihematomal neurons, and accelerated neurological recovery following collagenase-induced sICH. The integration of histological, immunological, and functional outcomes provides strong evidence supporting Kv1.3 as a promising therapeutic target for early intervention after hemorrhagic stroke. Although the study was limited to male mice, was restricted to a 7-day observation period, and lacked detailed downstream mechanistic analyses, the findings establish a robust preclinical foundation for future investigations into long-term functional recovery, molecular signaling pathways, and translational evaluation of Kv1.3-targeted therapies in sICH.
