Background and Aim: Amyloidosis is characterized by extracellular deposition of misfolded fibrillar proteins that can disrupt tissue architecture and function. Although amyloid deposition has been extensively investigated in several organs, testicular involvement remains poorly characterized. This study investigated whether sodium caseinate (CasNa) administration induces progressive amyloid deposition in the testes of adult male rats and characterized the associated histopathological, histochemical, and β-amyloid (Aβ) immunohistochemical changes. Materials and Methods: Twenty healthy adult male Albino Wistar rats (Rattus norvegicus) were randomly allocated to control and treatment groups (n = 10/group). Controls received 3.0 mL/kg intraperitoneal distilled water, whereas treated rats received 3.0 mL/kg body weight of 8% (w/v) CasNa. Five rats from each group were euthanized on days 25 and 60. Testicular tissues were evaluated using hematoxylin and eosin staining, histomorphometric measurement of seminiferous tubule diameter, and Congo red staining for amyloid-associated deposits. Immunohistochemistry was performed to assess Aβ immunoreactivity. Results: CasNa administration induced progressive testicular lesions. On day 25, treated rats showed seminiferous tubule alterations, vascular congestion and vasculitis, germinal epithelial detachment, impaired spermatogenesis, and proteinaceous deposition in the interstitium. By day 60, severe necrosis, loss of normal testicular architecture, and mononuclear inflammatory cell infiltration were evident. Seminiferous tubule diameter decreased significantly in treated rats, with a greater reduction on day 60 (241.4 ± 9.664) than on day 25 (301.7 ± 4.597) (p < 0.0001). Congo red staining demonstrated extracellular amyloid-associated deposits predominantly in the interstitial tissue and along seminiferous tubule basement membranes. Aβ immunoreactivity was absent in controls but was moderate on day 25 (30.42 ± 6.870) and intense on day 60 (61.34 ± 6.870) in treated rats. Immunoreactivity increased significantly in both treated groups compared with controls and was significantly greater on day 60 than on day 25 (p < 0.0001). Conclusion: Chronic CasNa administration induces progressive testicular amyloid-associated deposition, substantial structural damage, and increasing Aβ immunoreactivity in adult male rats. These findings support this experimental model for investigating testicular amyloidosis; however, Congo red staining and Aβ immunoreactivity alone don’t conclusively establish the biochemical identity of the deposited protein. Therefore, complementary molecular and biochemical approaches are required to confirm the identity of the deposited amyloid protein and determine its effects on male reproductive function. Keywords: amyloid deposition, β-amyloid, Congo red, histopathology, immunohistochemistry, sodium caseinate, testicular amyloidosis, Rattus norvegicus.
Background and Aim: Antimicrobial resistance (AMR) is a major global health concern affecting humans, animals, and the environment. Although surveillance has traditionally focused on healthcare settings and livestock, wildlife is increasingly recognized as a potential reservoir, carrier, and indicator of antimicrobial-resistant bacteria and resistance genes. This review examines the ecological factors, movement patterns, and transmission pathways influencing AMR in wildlife, with particular emphasis on potential spillover to humans and domestic animals. Wildlife can acquire resistant microorganisms through contaminated water, soil, food, agricultural runoff, wastewater, and interactions with humans and livestock. Habitat fragmentation, urbanization, agricultural expansion, feeding behavior, and predator–prey relationships further influence exposure and opportunities for bacterial transmission. Local movements and long-distance migration may contribute to the geographical dissemination of resistant bacteria and resistance genes between ecosystems. Evidence from different wildlife populations has identified extended-spectrum β-lactamase-producing Escherichia coli in migratory birds, methicillin-resistant Staphylococcus aureus in wild mammals, and fluoroquinolone-resistant bacteria in primates inhabiting areas with frequent human contact. Marine and aquatic wildlife may also contribute to AMR circulation through exposure to wastewater, pharmaceutical residues, and other environmental contaminants. Horizontal gene transfer involving plasmids, integrons, and transposons further contribute to the persistence and dissemination of resistance genes within microbial communities. Humans may be exposed through direct contact, wildlife handling, hunting, and indirect exposure to contaminated water, soil, and food. However, detecting resistant bacteria in wildlife does not necessarily establish the direction or occurrence of transmission to humans. Key challenges include limited wildlife surveillance, inconsistent sampling methods, insufficient ecological data, and difficulty identifying transmission sources. Integrating wildlife monitoring with genomic analysis, environmental surveillance, and animal movement studies could improve understanding of AMR distribution and transmission risks. A coordinated One Health approach involving human health, veterinary, environmental, and wildlife sectors is essential to better characterize potential transmission pathways, strengthen surveillance, and develop effective strategies to reduce AMR spread across interconnected ecosystems. Keywords: antimicrobial resistance, environmental contamination, horizontal gene transfer, migratory birds, One Health, resistance genes, wildlife reservoirs, zoonotic spillover.
Background and Aim: Antimalarial drug resistance highlights the need to identify alternative bioactive compounds. Azadirachta indica (neem) contains phytochemicals with potential antiplasmodial properties, although their molecular interactions remain incompletely characterized. This study aimed to characterize the metabolites of A. indica leaf extract, evaluate its antimalarial activity against Plasmodium berghei, and investigate predicted interactions between selected metabolites and parasite-associated proteins. Materials and Methods: Ethanolic leaf extract was characterized using liquid chromatography–high-resolution mass spectrometry (LC-HRMS). Twenty-five male BALB/c mice infected with chloroquine-sensitive P. berghei ANKA were randomly assigned to five groups (n = 5). Three groups received oral extract doses of 250, 500, or 750 mg/kg body weight for four consecutive days. Artesunate (2.4 mg/kg) and distilled water served as positive and negative controls, respectively. Parasitemia suppression was evaluated using a modified Peters' 4-day suppressive test. Selected metabolites underwent molecular docking against Plasmodium falciparum dihydrofolate reductase–thymidylate synthase, chloroquine resistance transporter, and plasmepsin II, alongside exploratory host-response targets caspase-3 and Bcl-2. Molecular dynamics simulations (100 ns) and binding free-energy calculations assessed predicted interaction stability. Results: LC-HRMS tentatively identified 33 metabolites, predominantly flavonols and flavonol glycosides. Scopoletin exhibited the greatest relative abundance, followed by rutin, trifolin, isoquercetin, quercetin 3-β-D-glucoside, and morin. Parasitemia suppression reached 78.11%, 82.08%, and 76.83% at 250, 500, and 750 mg/kg, respectively, compared with 95.75% for artesunate. The highest numerical suppression occurred at 500 mg/kg. Differences among extract-treated and artesunate groups were not statistically significant (p > 0.05), although therapeutic equivalence was not established. Computational analyses identified favorable predicted interactions involving catechin, luteolin, and 2-coumaroylquinic acid, particularly luteolin with the chloroquine resistance transporter. Conclusion: A. indica leaf extract demonstrated preliminary antimalarial activity against chloroquine-sensitive P. berghei. Integrated metabolite profiling and computational analyses identified candidate compounds warranting further investigation. Experimental target validation, individual-compound testing, and evaluation against drug-resistant parasites are necessary to establish mechanisms and therapeutic relevance. Keywords: Azadirachta indica, antimalarial activity, liquid chromatography–high-resolution mass spectrometry, molecular docking, molecular dynamics, neem leaf extract, Plasmodium berghei, phytochemical profiling.
Background and Aim: Repeated high-intensity exercise may alter reproductive hormone concentrations and compromise semen quality in performance stallions. L-carnitine participates in mitochondrial energy metabolism and has been investigated for its potential reproductive benefits. However, evidence regarding systemic L-carnitine supplementation in exercised Warmblood stallions remains limited. This study aimed to evaluate the effects of intramuscular L-carnitine administration on reproductive hormone concentrations and the quality of fresh and chilled semen in Warmblood stallions subjected to repeated exercise. Materials and Methods: Two clinically healthy, proven fertile Warmblood stallions were enrolled in a preliminary randomized-sequence crossover study. Each stallion underwent three experimental conditions: no exercise or supplementation (negative control), repeated exercise with saline placebo (positive control), and repeated exercise with intramuscular L-carnitine supplementation (6 mg/kg body weight, twice weekly). Each treatment period lasted 8 weeks, with an 8-week washout between periods. Serum luteinizing hormone, also termed interstitial cell-stimulating hormone, and testosterone concentrations were measured using enzyme-linked immunosorbent assays. Fresh semen was evaluated for macroscopic characteristics, progressive motility, viability, and plasma membrane integrity. Chilled semen was assessed at 0, 24, 48, and 72 h of storage. Results: Exercise with placebo was associated with lower reproductive hormone concentrations and poorer fresh semen quality than the negative control. L-carnitine supplementation was associated with higher luteinizing hormone (1.79 ± 0.28 vs. 1.12 ± 0.14 ng/mL) and testosterone (1.91 ± 0.26 vs. 1.43 ± 0.12 ng/mL) concentrations than exercise with placebo. Fresh semen progressive motility (73.40 ± 4.68% vs. 60.40 ± 1.47%), viability (80.00 ± 3.74% vs. 68.50 ± 1.77%), and plasma membrane integrity (64.10 ± 3.97% vs. 55.30 ± 1.10%) were also higher during supplementation. At 72 h of chilled storage, progressive motility was 36.92 ± 1.01% with supplementation and 34.79 ± 0.95% with placebo. Semen quality declined during storage under all conditions, with broadly similar proportional reductions. Conclusion: Intramuscular L-carnitine supplementation was associated with more favorable reproductive hormone concentrations and semen quality during repeated exercise. However, including only two stallions limits statistical inference and generalizability. Larger, controlled studies are needed before establishing practical breeding recommendations. Keywords: chilled semen, exercise-induced stress, health, L-carnitine, luteinizing hormone, sperm motility, testosterone, Warmblood stallions.
Background and Aim: Plant-derived compounds have attracted interest as potential complementary approaches to feline glycemic management. However, evidence regarding the metabolic effects of Paederia foetida in cats remains limited. This study aimed to characterize the phenolic and flavonoid contents and in vitro antioxidant activity of P. foetida leaf extract and evaluate its effects on glycemic and lipid parameters in cats subjected to a mild dietary carbohydrate challenge. Materials and Methods: Mature P. foetida leaves were extracted using 70% ethanol. Total phenolic and flavonoid contents were determined using colorimetric assays, and antioxidant activity was assessed using the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. Eighteen clinically healthy cats participated in a non-randomized, one-sequence crossover study. Following control diet and high-carbohydrate-diet phases, all cats received glipizide (2.5 mg/cat/day) for 4 weeks, underwent a 4-week washout, and subsequently received P. foetida extract (500 mg/cat/day) for 4 weeks. Fasting blood glucose, intravenous glucose tolerance, serum triglycerides, and total cholesterol were evaluated. Results: Total phenolic and flavonoid contents were 2.50 ± 0.33 mg gallic acid equivalents/g extract and 1.69 ± 0.50 mg quercetin equivalents/g extract, respectively. The extract exhibited relatively weak antioxidant activity, with a half-maximal inhibitory concentration of 2,000.00 ± 1.88 μg/mL. The high-carbohydrate diet increased fasting blood glucose from 85.3 ± 3.8 to 98.3 ± 4.3 mg/dL. Extract administration reduced fasting glucose to 88.1 ± 3.0 mg/dL (p < 0.05), whereas glipizide reduced it to 52.9 ± 7.7 mg/dL. Glucose tolerance improved following extract administration, although the response was less pronounced than with glipizide. Serum triglycerides were lower during extract treatment than during glipizide treatment, whereas the extract did not significantly reduce cholesterol relative to the high-carbohydrate-diet condition. Conclusion: P. foetida leaf extract was associated with modest improvements in glycemic parameters in cats experiencing mild diet-induced blood glucose elevation. However, its antioxidant activity was limited, and its glucose-lowering effect was smaller than that of glipizide. Randomized studies that optimize dosing, standardize phytochemical characterization, and include longer-term monitoring are needed before establishing clinical applicability. Keywords: antioxidant activity, feline glucose metabolism, glycemic regulation, high-carbohydrate diet, lipid profile, Paederia foetida, plant extract, sequential crossover study.
