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16 pages, 1185 KiB  
Article
Hematological, Enzymatic, and Endocrine Response to Intense Exercise in Lidia Breed Cattle During the Roping Bull Bullfighting Celebration
by Julio Sedeño, Salvador Ruiz, Germán Martín and Juan Carlos Gardón
Animals 2025, 15(15), 2303; https://doi.org/10.3390/ani15152303 (registering DOI) - 6 Aug 2025
Abstract
The Lidia cattle breed is featured in several traditional popular bullfighting festivals throughout Spain, including the “Toro de Cuerda” event, in which the animals are subjected to intense physical exercise. However, the physiological impact and welfare implications of these activities remain poorly characterized. [...] Read more.
The Lidia cattle breed is featured in several traditional popular bullfighting festivals throughout Spain, including the “Toro de Cuerda” event, in which the animals are subjected to intense physical exercise. However, the physiological impact and welfare implications of these activities remain poorly characterized. This study aimed to evaluate the stress response and muscle damage in Lidia breed bulls during roping bull celebrations through comprehensive blood analysis. Blood samples were collected from 53 adult male Lidia bulls before and after a standardized 45 min continuous running exercise during traditional roping bull events in four Spanish autonomous regions. Hematological parameters, muscle enzymes (creatine kinase, lactate dehydrogenase, lactate), and stress hormones (cortisol and ACTH) were analyzed. Significant increases (p < 0.05) were observed in leukocytes, lymphocytes, monocytes, eosinophils, neutrophils, erythrocytes, hematocrit, hemoglobin, and post-exercise platelets. Muscle enzymes showed marked elevations, with creatine kinase increasing up to 10-fold above baseline values. Stress hormones, cortisol and ACTH, also demonstrated significant increases. Despite the magnitude of these changes, all parameters remained within established reference ranges for the bovine species. This study provides the first physiological assessment of Lidia cattle during popular bullfighting celebrations, establishing baseline data for evidence-based welfare evaluation and management protocols. Full article
(This article belongs to the Section Cattle)
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15 pages, 2022 KiB  
Article
Dual-Emission Au-Ag Nanoclusters with Enhanced Photoluminescence and Thermal Sensitivity for Intracellular Ratiometric Nanothermometry
by Helin Liu, Zhongliang Zhou, Zhiwei Wang, Jianhai Wang, Yu Wang, Lu Huang, Tianhuan Guo, Rongcheng Han and Yuqiang Jiang
Biosensors 2025, 15(8), 510; https://doi.org/10.3390/bios15080510 (registering DOI) - 6 Aug 2025
Abstract
We report the development of highly luminescent, bovine serum albumin (BSA)-stabilized gold–silver bimetallic nanoclusters (Au-AgNCs@BSA) as a novel platform for high-sensitivity, ratiometric intracellular temperature sensing. Precise and non-invasive temperature sensing at the nanoscale is crucial for applications ranging from intracellular thermogenesis monitoring to [...] Read more.
We report the development of highly luminescent, bovine serum albumin (BSA)-stabilized gold–silver bimetallic nanoclusters (Au-AgNCs@BSA) as a novel platform for high-sensitivity, ratiometric intracellular temperature sensing. Precise and non-invasive temperature sensing at the nanoscale is crucial for applications ranging from intracellular thermogenesis monitoring to localized hyperthermia therapies. Traditional luminescent thermometric platforms often suffer from limitations such as high cytotoxicity and low photostability. Here, we synthesized Au-AgNCs@BSA via a one-pot aqueous reaction, achieving significantly enhanced photoluminescence quantum yields (PL QYs, up to 18%) and superior thermal responsiveness compared to monometallic counterparts. The dual-emissive Au-AgNCs@BSA exhibit a linear ratiometric fluorescence response to temperature fluctuations within the physiological range (20–50 °C), enabling accurate and concentration-independent thermometry in live cells. Time-resolved PL and Arrhenius analyses reveal two distinct emissive states and a high thermal activation energy (Ea = 199 meV), indicating strong temperature dependence. Silver doping increases radiative decay rates while maintaining low non-radiative losses, thus amplifying fluorescence intensity and thermal sensitivity. Owing to their small size, excellent photostability, and low cytotoxicity, these nanoclusters were applied to non-invasive intracellular temperature mapping, presenting a promising luminescent nanothermometer for real-time cellular thermogenesis monitoring and advanced bioimaging applications. Full article
(This article belongs to the Section Nano- and Micro-Technologies in Biosensors)
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15 pages, 1040 KiB  
Article
Alcalase Specificity by Different Substrate Proteins Under Different Conditions: The Enzyme Immobilization on Carrageenan Beads Strongly Affects the pH/Activity Curve Depending on the Substrate Protein
by Alan Portal D’Almeida, Pedro Abellanas-Perez, Luciana Rocha Barros Gonçalves, Tiago Lima de Albuquerque, Ivanildo José da Silva Junior and Roberto Fernandez-Lafuente
Catalysts 2025, 15(8), 750; https://doi.org/10.3390/catal15080750 - 5 Aug 2025
Abstract
Alcalase was immobilized–stabilized on carrageenan beads following a previously described protocol. Then, the activities of free and immobilized enzymes were compared using different protein substrates (casein, (CS), bovine serum albumin (BSA), or hemoglobin (HG)) at different pH values and temperatures. The observed activity [...] Read more.
Alcalase was immobilized–stabilized on carrageenan beads following a previously described protocol. Then, the activities of free and immobilized enzymes were compared using different protein substrates (casein, (CS), bovine serum albumin (BSA), or hemoglobin (HG)) at different pH values and temperatures. The observed activity depended on the substrate protein and enzyme formulation used. The highest enzyme activity could be observed at pHs 5, 7, or 10, depending on the substrate protein and the Alcalase formulation. The effect of the temperature at these pHs on the activity versus the different substrate proteins showed a common pattern. At low temperatures, the immobilized enzyme presented higher (mainly at acidic-neutral pH values and using BSA) or similar specific activity than the free enzyme. At temperatures near the optimal for the free enzyme, it became the most active, while at higher temperatures, the immobilized enzyme recovered the lead, although differences in the optimal temperature were not very significant. This may be explained by the lower mobility of the immobilized–stabilized enzyme. The immobilized enzyme could be much more active than the free enzyme or slightly less active, even using mild conditions, depending on the substrate protein, pH, and temperature used to determine the enzyme activity. Full article
(This article belongs to the Section Biocatalysis)
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14 pages, 1669 KiB  
Article
Guinea Pig X Virus Is a Gammaherpesvirus
by Vy Ngoc Yen Truong, Robert Ellis and Brent A. Stanfield
Viruses 2025, 17(8), 1084; https://doi.org/10.3390/v17081084 - 5 Aug 2025
Abstract
The Guinea Pig X Virus (GPXV), a newly identified gammaherpesvirus, provides an opportunity to study viral evolution and host–virus dynamics. This study characterizes the GPXV genome and investigates its phylogenetic relationships and divergence from related viruses through comparative genomic and phylogenetic analyses. Virus [...] Read more.
The Guinea Pig X Virus (GPXV), a newly identified gammaherpesvirus, provides an opportunity to study viral evolution and host–virus dynamics. This study characterizes the GPXV genome and investigates its phylogenetic relationships and divergence from related viruses through comparative genomic and phylogenetic analyses. Virus propagation was conducted in Vero cells, followed by genomic DNA extraction and pan-herpesvirus nested PCR. Sanger sequencing filled gaps in the initial genome assembly, and whole-genome sequencing was performed using the Illumina MiSeq platform. Phylogenetic analyses focused on ORF8 (glycoprotein B), ORF9 (DNA polymerase catalytic subunit), ORF50 (RTA: replication and transcription activator), and ORF73 (LANA: latency-associated nuclear antigen). Results showed that GPXV ORFs showed variable evolutionary relationships with other gammaherpesviruses, including divergence from primate-associated viruses and clustering with bovine and rodent viruses. In addition to phylogenetics, a comprehensive comparative analysis of protein-coding genes between GPXV and the previously described Guinea Pig Herpes-Like Virus (GPHLV) revealed divergence. Twenty-four non-ORF genomic features were unique to GPXV, while 62 shared ORFs exhibited low to high sequence divergence. These findings highlight GPXV’s distinct evolutionary trajectory and its potential role as a model for studying host-specific adaptations and gammaherpesvirus diversity. Full article
(This article belongs to the Special Issue Animal Herpesvirus 2025)
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11 pages, 486 KiB  
Article
Multi-Mycotoxin Contamination of Concentrates Fed to Dairy Calves in Southeast Brazil: A Case Report
by Rogério D’Antonio Pires, Aline Moreira Borowsky, Tobias Alves e Silva, Giovanna Canela Ruiz Castro Evangelista, Carla Maris Machado Bittar and Carlos Humberto Corassin
Dairy 2025, 6(4), 44; https://doi.org/10.3390/dairy6040044 - 4 Aug 2025
Viewed by 81
Abstract
The diet of dairy calves can be contaminated with mycotoxins, posing a potential risk to animal health. This case study report aimed to make the first assessment of the presence of multiple mycotoxins in concentrates fed to dairy calves in Brazil. A total [...] Read more.
The diet of dairy calves can be contaminated with mycotoxins, posing a potential risk to animal health. This case study report aimed to make the first assessment of the presence of multiple mycotoxins in concentrates fed to dairy calves in Brazil. A total of 19 concentrate samples intended for dairy calves were analyzed using liquid chromatography coupled with mass spectrometry. Aflatoxins, deoxynivalenol, and T-2 toxin were not detected in any samples, whereas fumonisins B1 (FB1) and B2 (FB2) were present in 100% of the samples, with mean concentrations of 2750.1 μg/kg and 834.9 μg/kg, respectively. Zearalenone (ZEN) was detected in 36.8% of samples, with a mean concentration of 929.9 μg/kg. Significant correlations were observed between FB1 and FB2 (ρ = 0.978; p < 0.001) and between FB2 and ZEN (ρ = 0.735; p = 0.05). While the physical form of the concentrate did not influence (p > 0.05) mycotoxin concentrations, a trend was observed for FB1 (ρ = −0.417; p = 0.07) and FB2 (ρ = −0.395; p = 0.09). These findings highlight the frequent occurrence of Fusarium mycotoxins, likely due to pre-harvest contamination, emphasizing the potential risk of additive or synergistic effects in dairy calves. Full article
(This article belongs to the Section Dairy Animal Nutrition and Welfare)
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12 pages, 2807 KiB  
Article
Evaluation of Hydroxyapatite–β-Tricalcium Phosphate Collagen Composites for Socket Preservation in a Canine Model
by Dong Woo Kim, Donghyun Lee, Jaeyoung Ryu, Min-Suk Kook, Hong-Ju Park and Seunggon Jung
J. Funct. Biomater. 2025, 16(8), 286; https://doi.org/10.3390/jfb16080286 - 3 Aug 2025
Viewed by 218
Abstract
This study aimed to compare the performance of three hydroxyapatite–β-tricalcium phosphate (HA–β-TCP) collagen composite grafts in a canine model for extraction socket preservation. Eight mongrel dogs underwent atraumatic bilateral mandibular premolar extraction, and sockets were randomly grafted with HBC28 (20% high-crystalline HA, 80% [...] Read more.
This study aimed to compare the performance of three hydroxyapatite–β-tricalcium phosphate (HA–β-TCP) collagen composite grafts in a canine model for extraction socket preservation. Eight mongrel dogs underwent atraumatic bilateral mandibular premolar extraction, and sockets were randomly grafted with HBC28 (20% high-crystalline HA, 80% β-TCP bovine collagen), HBC37 (30% HA, 70% β-TCP, bovine collagen), or HPC64 (60% HA, 40% β-TCP, porcine collagen). Grafts differed in their HA–β-TCP ratio and collagen origin and content. Animals were sacrificed at 4 and 12 weeks, and the healing sites were evaluated using micro-computed tomography (micro-CT) and histological analysis. At 12 weeks, all groups showed good socket maintenance with comparable new bone formation. However, histological analysis revealed that HBC28 had significantly higher residual graft volume, while HPC64 demonstrated more extensive graft resorption. Histomorphometric analysis confirmed these findings, with statistically significant differences in residual graft area and bone volume fraction. No inflammatory response or adverse tissue reactions were observed in any group. These results suggest that all three HA–β-TCP collagen composites are biocompatible and suitable for socket preservation, with varying resorption kinetics influenced by graft composition. Selection of graft material may thus be guided by the desired rate of replacement by new bone. Full article
(This article belongs to the Special Issue Biomechanical Studies and Biomaterials in Dentistry)
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14 pages, 265 KiB  
Article
Bovine Leptospirosis: Serology, Isolation, and Risk Factors in Dairy Farms of La Laguna, Mexico
by Alejandra María Pescador-Gutiérrez, Jesús Francisco Chávez-Sánchez, Lucio Galaviz-Silva, Juan José Zarate-Ramos, José Pablo Villarreal-Villarreal, Sergio Eduardo Bernal-García, Uziel Castillo-Velázquez, Rubén Cervantes-Vega and Ramiro Avalos-Ramirez
Life 2025, 15(8), 1224; https://doi.org/10.3390/life15081224 - 2 Aug 2025
Viewed by 188
Abstract
Leptospirosis is a globally significant zoonosis affecting animal health, productivity, and the environment. While typically associated with tropical climates, its persistence in semi-arid regions such as La Laguna, Mexico—characterized by low humidity, high temperatures, and limited water sources—remains poorly understood. Although these adverse [...] Read more.
Leptospirosis is a globally significant zoonosis affecting animal health, productivity, and the environment. While typically associated with tropical climates, its persistence in semi-arid regions such as La Laguna, Mexico—characterized by low humidity, high temperatures, and limited water sources—remains poorly understood. Although these adverse environmental conditions theoretically limit the survival of Leptospira, high livestock density and synanthropic reservoirs (e.g., rodents) may compensate, facilitating transmission. In this cross-sectional study, blood sera from 445 dairy cows (28 herds: 12 intensive [MI], 16 semi-intensive [MSI] systems) were analyzed via microscopic agglutination testing (MAT) against 10 pathogenic serovars. Urine samples were cultured for active Leptospira detection. Risk factors were assessed through epidemiological surveys and multivariable analysis. This study revealed an overall apparent seroprevalence of 27.0% (95% CI: 22.8–31.1), with significantly higher rates in MSI (54.1%) versus MI (12.2%) herds (p < 0.001) and an estimated true seroprevalence of 56.3% (95% CI: 50.2–62.1) in MSI and 13.1% (95% CI: 8.5–18.7) in MI herds (p < 0.001). The Sejroe serogroup was isolated from urine in both systems, confirming active circulation. In MI herds, rodent presence (OR: 3.6; 95% CI: 1.6–7.9) was identified as a risk factor for Leptospira seropositivity, while first-trimester abortions (OR:10.1; 95% CI: 4.2–24.2) were significantly associated with infection. In MSI herds, risk factors associated with Leptospira seropositivity included co-occurrence with hens (OR: 2.8; 95% CI: 1.5–5.3) and natural breeding (OR: 2.0; 95% CI: 1.1–3.9), whereas mastitis/agalactiae (OR: 2.8; 95% CI: 1.5–5.2) represented a clinical outcome associated with seropositivity. Despite semi-arid conditions, Leptospira maintains transmission in La Laguna, particularly in semi-intensive systems. The coexistence of adapted (Sejroe) and incidental serogroups underscores the need for targeted interventions, such as rodent control in MI systems and poultry management in MSI systems, to mitigate both zoonotic and economic impacts. Full article
(This article belongs to the Section Animal Science)
22 pages, 598 KiB  
Article
Re-Consider the Lobster: Animal Lives in Protein Supply Chains
by Karl T. Ulrich
Sustainability 2025, 17(15), 7034; https://doi.org/10.3390/su17157034 - 2 Aug 2025
Viewed by 120
Abstract
Animal protein production represents a complex system of lives transformed into nutrition, with profound ethical and environmental implications. This study provides a quantitative analysis of animal lives required to produce human-consumable protein across major food production systems. Categorizing animal lives based on cognitive [...] Read more.
Animal protein production represents a complex system of lives transformed into nutrition, with profound ethical and environmental implications. This study provides a quantitative analysis of animal lives required to produce human-consumable protein across major food production systems. Categorizing animal lives based on cognitive complexity and accounting for all lives involved in production, including direct harvests, reproductive animals, and feed species, reveals dramatic variations in protein efficiency. The analysis considers two categories of animal life: complex-cognitive lives (e.g., mammals, birds, cephalopods) and pain-capable lives (e.g., fish, crustaceans). Calculating protein yield per life demonstrates efficiency differences spanning more than five orders of magnitude, from 2 g per complex-cognitive life for baby octopus to 390,000 g per life for bovine dairy systems. Key findings expose disparities between terrestrial and marine protein production. Terrestrial systems involving mammals and birds show higher protein yields and exclusively involve complex-cognitive lives, while marine systems rely predominantly on pain-capable lives across complex food chains. Dairy production emerges as the most efficient system. Aquaculture systems reveal complex dynamics, with farmed carnivorous fish requiring hundreds of feed fish lives to produce protein, compared to omnivorous species that demonstrate improved efficiency. Beyond quantitative analysis, this research provides a framework for understanding the ethical and ecological dimensions of protein production, offering insights for potential systemic innovations. Full article
(This article belongs to the Section Sustainable Food)
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10 pages, 868 KiB  
Article
The Response of Cell Cultures to Nutrient- and Serum-Induced Changes in the Medium
by Marijana Leventić, Katarina Mišković Špoljarić, Karla Vojvodić, Nikolina Kovačević, Marko Obradović and Teuta Opačak-Bernardi
Sci 2025, 7(3), 105; https://doi.org/10.3390/sci7030105 - 2 Aug 2025
Viewed by 178
Abstract
Cell culture models are of central importance for the investigation of cellular metabolism, proliferation and stress responses. In this study, the effects of different concentrations of glucose (1 g/L vs. 4.5 g/L) and fetal bovine serum (FBS; 5%, 10%, 15%) on viability, mitochondrial [...] Read more.
Cell culture models are of central importance for the investigation of cellular metabolism, proliferation and stress responses. In this study, the effects of different concentrations of glucose (1 g/L vs. 4.5 g/L) and fetal bovine serum (FBS; 5%, 10%, 15%) on viability, mitochondrial function and autophagy are investigated in four human cell lines: MRC-5, HeLa, Caco-2 and SW-620. Cells were cultured in defined media for 72 h, and viability was assessed by LDH release, mitochondrial membrane potential using Rhodamine 123, ATP content by luminescence and autophagy activity by dual fluorescence staining. The results showed that HeLa and SW-620 cancer cells exhibited increased proliferation and mitochondrial activity under high glucose conditions, while low glucose media resulted in decreased ATP content and increased membrane permeability in HeLa cells. MRC-5 fibroblasts and Caco-2 cells showed greater resilience to nutrient stress, with minimal changes in LDH release and consistent proliferation. Autophagy was activated under all conditions, with a significant increase only in selected cell-medium combinations. These results highlight the importance of medium composition in influencing cellular bioenergetics and stress responses, which has implications for cancer research, metabolic disease modelling and the development of serum-free culture systems for regenerative medicine. Full article
(This article belongs to the Section Biology Research and Life Sciences)
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19 pages, 1134 KiB  
Article
Application of Animal- and Plant-Derived Coagulant in Artisanal Italian Caciotta Cheesemaking: Comparison of Sensory, Biochemical, and Rheological Parameters
by Giovanna Lomolino, Stefania Zannoni, Mara Vegro and Alberto De Iseppi
Dairy 2025, 6(4), 43; https://doi.org/10.3390/dairy6040043 - 1 Aug 2025
Viewed by 88
Abstract
Consumer interest in vegetarian, ethical, and clean-label foods is reviving the use of plant-derived milk coagulants. Cardosins from Cynara cardunculus (“thistle”) are aspartic proteases with strong clotting activity, yet their technological impact in cheese remains under-explored. This study compared a commercial thistle extract [...] Read more.
Consumer interest in vegetarian, ethical, and clean-label foods is reviving the use of plant-derived milk coagulants. Cardosins from Cynara cardunculus (“thistle”) are aspartic proteases with strong clotting activity, yet their technological impact in cheese remains under-explored. This study compared a commercial thistle extract (PC) with traditional bovine rennet rich in chymosin (AC) during manufacture and 60-day ripening of Caciotta cheese. Classical compositional assays (ripening index, texture profile, color, solubility) were integrated with scanning electron microscopy, three-dimensional surface reconstruction, and descriptive sensory analysis. AC cheeses displayed slower but sustained proteolysis, yielding a higher and more linear ripening index, softer body, greater solubility, and brighter, more yellow appearance. Imaging revealed a continuous protein matrix with uniformly distributed, larger pores, consistent with a dairy-like sensory profile dominated by milky and umami notes. Conversely, PC cheeses underwent rapid early proteolysis that plateaued, producing firmer, chewier curds with lower solubility and darker color. Micrographs showed a fragmented matrix with smaller, heterogeneous pores; sensory evaluation highlighted vegetal, bitter, and astringent attributes. The data demonstrate that thistle coagulant can successfully replace animal rennet but generates cheeses with distinct structural and sensory fingerprints. The optimization of process parameters is therefore required when targeting specific product styles. Full article
(This article belongs to the Section Milk Processing)
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22 pages, 7580 KiB  
Article
Bacterial and Physicochemical Dynamics During the Vermicomposting of Bovine Manure: A Comparative Analysis of the Eisenia fetida Gut and Compost Matrix
by Tania Elizabeth Velásquez-Chávez, Jorge Sáenz-Mata, Jesús Josafath Quezada-Rivera, Rubén Palacio-Rodríguez, Gisela Muro-Pérez, Alan Joel Servín-Prieto, Mónica Hernández-López, Pablo Preciado-Rangel, María Teresa Salazar-Ramírez, Juan Carlos Ontiveros-Chacón and Cristina García-De la Peña
Microbiol. Res. 2025, 16(8), 177; https://doi.org/10.3390/microbiolres16080177 - 1 Aug 2025
Viewed by 109
Abstract
Vermicomposting is a sustainable biotechnological process that transforms organic waste through the synergistic activity of earthworms, such as Eisenia fetida, and their associated microbiota. This study evaluated bacterial and physicochemical dynamics during the vermicomposting of bovine manure by analyzing the microbial composition [...] Read more.
Vermicomposting is a sustainable biotechnological process that transforms organic waste through the synergistic activity of earthworms, such as Eisenia fetida, and their associated microbiota. This study evaluated bacterial and physicochemical dynamics during the vermicomposting of bovine manure by analyzing the microbial composition of the substrate and the gut of E. fetida at three time points (weeks 0, 6, and 12). The V3–V4 region of the 16S rRNA gene was sequenced, and microbial diversity was characterized using QIIME2. Significant differences in alpha diversity (observed features, Shannon index, and phylogenetic diversity) and beta diversity indicated active microbial succession. Proteobacteria, Bacteroidota, and Actinobacteriota were the dominant phyla, with abundances varying across habitats and over time. A significant enrichment of Proteobacteria, Bacteroidota, and the genera Chryseolinea, Flavobacterium, and Sphingomonas was observed in the manure treatments. In contrast, Actinobacteriota, Firmicutes, and the genera Methylobacter, Brevibacillus, Enhygromyxa, and Bacillus, among others, were distinctive of the gut samples and contributed to their dissimilarity from the manure treatments. Simultaneously, the physicochemical parameters indicated progressive substrate stabilization and nutrient enrichment. Notably, the organic matter and total organic carbon contents decreased (from 79.47% to 47.80% and from 46.10% to 27.73%, respectively), whereas the total nitrogen content increased (from 1.70% to 2.23%); these effects reduced the C/N ratio, which is a recognized indicator of maturity, from 27.13 to 12.40. The macronutrient contents also increased, with final values of 1.41% for phosphorus, 1.50% for potassium, 0.89% for magnesium, and 2.81% for calcium. These results demonstrate that vermicomposting modifies microbial communities and enhances substrate quality, supporting its use as a biofertilizer for sustainable agriculture, soil restoration, and agrochemical reduction. Full article
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17 pages, 1353 KiB  
Article
Inhibition of Human Coronavirus 229E by Lactoferrin-Derived Peptidomimetics
by Maria Carmina Scala, Magda Marchetti, Martina Landi, Marialuigia Fantacuzzi, Fabiana Superti, Mariangela Agamennone, Pietro Campiglia and Marina Sala
Pharmaceutics 2025, 17(8), 1006; https://doi.org/10.3390/pharmaceutics17081006 - 1 Aug 2025
Viewed by 237
Abstract
Background/Objectives: Viral respiratory infections have a significant impact on global health and the economy. While vaccines are effective in preventing infection, they might not be available or sufficient when used alone and must be complemented by specific therapeutic strategies. The development of new [...] Read more.
Background/Objectives: Viral respiratory infections have a significant impact on global health and the economy. While vaccines are effective in preventing infection, they might not be available or sufficient when used alone and must be complemented by specific therapeutic strategies. The development of new antiviral agents is increasingly important due to the continual emergence of novel respiratory pathogens. Previously we identified bovine lactoferrin (bLf)-derived tetrapeptides and peptidomimetics that showed potent in vitro activity against the influenza A virus in the picomolar range. Methods: Inspired by these results, in this study, we evaluated the antiviral potential of these compounds against HCoV-229E, a human coronavirus that can cause severe disease in immunocompromised individuals, using a compound repositioning approach. Results: Functional studies revealed that SK(N-Me)HS (3) interferes with viral entry and replication, while compound SNKHS (5) primarily blocks infection in the early stages. Biophysical analyses confirmed the occurrence of high-affinity binding to the viral spike protein, and computational studies suggested that the compounds target a region involved in conformational changes necessary for membrane fusion. Conclusions: These findings highlight these compounds as promising candidates for coronavirus entry inhibition and underscore the value of compound repurposing in antiviral development. Full article
(This article belongs to the Special Issue Peptides-Based Antiviral Agents)
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14 pages, 939 KiB  
Review
Revisiting Male Fertility in Livestock: The Case of Bull Sperm RNA
by Rene A. Ramírez-Sosa, Francisco J. Jahuey-Martínez, Monserrath Felix-Portillo and José A. Martínez-Quintana
Biology 2025, 14(8), 969; https://doi.org/10.3390/biology14080969 (registering DOI) - 1 Aug 2025
Viewed by 282
Abstract
To achieve the goals of productivity and sustainability across diverse livestock systems, reproductive factors play a pivotal role. Historically, reproductive research has primarily focused on females, as they are responsible for maintaining pregnancy and delivering offspring following oocyte fertilization. However, since the early [...] Read more.
To achieve the goals of productivity and sustainability across diverse livestock systems, reproductive factors play a pivotal role. Historically, reproductive research has primarily focused on females, as they are responsible for maintaining pregnancy and delivering offspring following oocyte fertilization. However, since the early 2000s, the biological significance of sperm RNAs has been increasingly recognized in various livestock species. These RNAs contribute both genetically and epigenetically at the time of fertilization and during early embryonic development. Multiple types of sperm RNA have been identified in bovine, porcine, ovine, buffalo, and caprine spermatozoa. Notably, transcriptomic profiling has shown potential to differentiate between high- and low-fertility males, even when conventional semen quality values appear normal in both groups. This opens the possibility for more accurate identification of highly fertile sires. Nevertheless, a definitive marker or set of markers has yet to be established, likely due to the transcriptome’s sensitivity to environmental conditions and to the variability in evaluation methodologies. Therefore, global scientific efforts should aim to establish standardized, robust protocols, as sperm RNA represents a promising avenue for enhancing the sustainability of animal production systems. Full article
(This article belongs to the Special Issue The Biology of Animal Reproduction)
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14 pages, 2882 KiB  
Article
Babesia bovis Enolase Is Expressed in Intracellular Merozoites and Contains B-Cell Epitopes That Induce Neutralizing Antibodies In Vitro
by Alma Cárdenas-Flores, Minerva Camacho-Nuez, Massaro W. Ueti, Mario Hidalgo-Ruiz, Angelina Rodríguez-Torres, Diego Josimar Hernández-Silva, José Guadalupe Gómez-Soto, Masahito Asada, Shin-ichiro Kawazu, Alma R. Tamayo-Sosa, Rocío Alejandra Ruiz-Manzano and Juan Mosqueda
Vaccines 2025, 13(8), 818; https://doi.org/10.3390/vaccines13080818 (registering DOI) - 31 Jul 2025
Viewed by 216
Abstract
Background: Bovine babesiosis, caused by the tick-borne apicomplexan parasite Babesia spp., is an economically significant disease that threatens the cattle industry worldwide. Babesia bovis is the most pathogenic species, leading to high morbidity and mortality in infected animals. One promising approach to [...] Read more.
Background: Bovine babesiosis, caused by the tick-borne apicomplexan parasite Babesia spp., is an economically significant disease that threatens the cattle industry worldwide. Babesia bovis is the most pathogenic species, leading to high morbidity and mortality in infected animals. One promising approach to vaccination against bovine babesiosis involves the use of multiple protective antigens, offering advantages over traditional live-attenuated vaccines. Tools such as immunobioinformatics and reverse vaccinology have facilitated the identification of novel antigens. Enolase, a “moonlighting” enzyme of the glycolytic pathway with demonstrated vaccine potential in other pathogens, has not yet been studied in B. bovis. Methods: In this study, the enolase gene from two B. bovis isolates was successfully identified and sequenced. The gene, consisting of 1366 base pairs, encodes a predicted protein of 438 amino acids. Its expression in intraerythrocytic parasites was confirmed by RT-PCR. Two peptides containing predicted B-cell epitopes were synthesized and used to immunize rabbits. Hyperimmune sera were then analyzed by ELISA, confocal microscopy, Western blot, and an in vitro neutralization assay. Results: The hyperimmune sera showed high antibody titers, reaching up to 1:256,000. Specific antibodies recognized intraerythrocytic merozoites by confocal microscopy and bound to a ~47 kDa protein in erythrocytic cultures of B. bovis as detected by Western blot. In the neutralization assay, antibodies raised against peptide 1 had no observable effect, whereas those targeting peptide 2 significantly reduced parasitemia by 71.99%. Conclusions: These results suggest that B. bovis enolase contains B-cell epitopes capable of inducing neutralizing antibodies and may play a role in parasite–host interactions. Enolase is therefore a promising candidate for further exploration as a vaccine antigen. Nonetheless, additional experimental studies are needed to fully elucidate its biological function and validate its vaccine potential. Full article
(This article belongs to the Special Issue Vaccines against Arthropods and Arthropod-Borne Pathogens)
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Article
Diversity and Seasonal Abundance of Culicoides (Diptera: Ceratopogonidae) in Tengchong County of Yunnan, China
by Yi-Nan Wang, Ying-Liang Duan, Zhan-Hong Li, Jia-Ming Deng, Xing-Nan Sun, Xue-Ying Shen, An-Xi Yang and Shi-Long Li
Insects 2025, 16(8), 780; https://doi.org/10.3390/insects16080780 - 30 Jul 2025
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Abstract
Culicoides (Diptera, Ceratopogonidae) are small biting midges and are known as vectors for many arboviruses, including bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV). Tengchong County of Yunnan Province, China, which borders Myanmar, has many private farms with goats, sheep, and cattle. [...] Read more.
Culicoides (Diptera, Ceratopogonidae) are small biting midges and are known as vectors for many arboviruses, including bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV). Tengchong County of Yunnan Province, China, which borders Myanmar, has many private farms with goats, sheep, and cattle. To estimate the risk of Culicoides-borne viral diseases such as bluetongue (BT) and epizootic hemorrhagic disease (EHD) in this area, an investigation of the diversity and abundance of Culicoides in Tengchong between May 2024 and April 2025 was performed. As a result, 70 collections totaling approximately 93,000 Culicoides were carried out at five farms (cattle + Asian buffaloes, goats, and sheep, respectively). Nineteen species were identified, and eight potential cryptic species were found. A total of 13 cox1 sequences and 4 28S sequences for 13 specimens were generated. The most dominant species were Obsoletus (44.1%), C. homotomus (23.3%), and C. arakawae (12.9%) at the bovine farm; C. tainanus (68.0%), C. orientalis (12.6%), and C. newsteadi (Asia) (6.3%) at the goat farm; and C. tainanus (73.6%), C. fenggangensis (7.3%), and C. sp. nr palpifer (6.3%) at the sheep farm. In this investigation, C. tainanus, Obsoletus, and C. orientalis were the most dominant potential BTV vectors, and the period between July and October may be the main period for epidemics of Culicoides-borne viruses in Tengchong. Full article
(This article belongs to the Section Insect Systematics, Phylogeny and Evolution)
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