Search Results (168)

The objective of this study was to evaluate the effect of various additives on the nutritional value and aerobic stability of safflower (Carthamus tinctorius L.) silages. Silages were prepared from whole safflower plants harvested 102 days after planting, which were chopped to a particle size of 2.0 ± 0.5 cm and fermented for 120 days in polyvinyl chloride microsilos (6” × 46 cm), evaluating the following treatments: (1) safflower silage (SS) without additives, (2) SS supplemented with Guanacaste tree (Enterolobium cyclocarpum) pod meal, (3) SS supplemented with corn meal, (4) SS supplemented with sorghum meal, (5) SS supplemented with molasses, (6) SS supplemented with homofermentative inoculant, and (7) SS supplemented with fermentative inoculant + molasses. Compared with SS without additives, the addition of all the evaluated additives increased (p < 0.0001) the crude protein content and the relative forage value, while simultaneously decreasing the pH in SS. In contrast, the use of Guanacaste tree pod meal, corn, and sorghum decreased (p < 0.0001) the neutral detergent fiber and acid detergent fiber contents, while simultaneously increasing (p < 0.0001) the in vitro digestibility of dry matter in SS. All the evaluated additives increased (p < 0.05) the aerobic stability of the SS, which broke 42 h after opening the microsilos, whereas the silage without additives broke at 30 h. In conclusion, the use of Guanacaste tree pod meal, corn, and sorghum as additives improves the nutritive value and aerobic stability of safflower silage. Full article

This study aimed to optimize the inoculation dosage and fermentation duration to enhance the protein content and reduce soluble oligosaccharides in soybean meal using Aspergillus oryzae and assessed its performance in dog food extrusion. A 3 × 5 factorial design was used to determine the optimal fermentation conditions. These conditions were applied to ferment soybean meal in bulk for nutritional analysis. Finally, the impact of fermentation on extrusion processing was assessed by formulating and extruding four diets: SBM (30% soybean meal), AMF (30% soybean meal with 1% Amaferm^®—A. oryzae biomass), FSBM (30% fermented soybean meal), and SPI (18% soy protein isolate). Diets were extruded with a single-screw extruder, and physical characteristics of kibbles, particle size distribution, and viscosity of raw mixes were analyzed. The optimal fermentation conditions were 1 × 10⁴ spore/g substrate for 36 h, which increased the crude protein content by 4.63% DM, methionine and cysteine total content by 0.15% DM, and eliminated sucrose, while significantly reducing stachyose, raffinose, and verbascose (95.22, 87.37, and 41.82%, respectively). The extrusion results showed that FSBM had intermediate specific mechanical energy (SME), in-barrel moisture requirements, and sectional expansion index (198.7 kJ/kg, 28.2%, and 1.80, respectively) compared with SBM (83.7 kJ/kg, 34.5%, and 1.30, respectively) and SPI (305.3 kJ/kg, 33.5%, and 2.55, respectively). The FSBM also exhibited intermediate particle size distribution and the least raw mix viscosity. These findings demonstrate that A. oryzae fermentation enhances the nutrient profile of soybean meal while improving extrusion efficiency and kibble quality, supporting its potential use as a sustainable pet food ingredient. Full article

The objective of this study was to evaluate the time-course of incubation for the potential preventative mitigation of megaviruses using Termin-8 (a formaldehyde-based product) and Finio (non-formaldehyde solution) from Anitox. Emiliania huxleyi virus (EhV), an algal surrogate for African swine fever virus (ASFV), was treated with the recommended concentrations of Termin-8 (0.1% to 0.3%) and Finio (0.05% to 0.2%), and both viability qPCR (V-qPCR) and standard PCR (S-qPCR) were used to quantify EhV concentrations at 1 h, 5 h, 24 h and day 7 post-inoculation. Overall, Finio, and to a lesser extent Termin-8, at their highest treatment concentrations, showed the greatest log reduction of 4.5 and 2 log₁₀ units, respectively, at 1 h post-inoculation. Although Termin-8 efficacy did not improve with time, due to its fixing of viral particles and rendering them non-infectious, treatment with Finio showed 100% viable viral inactivation (>5 log₁₀ reduction units) at the lowest concentration after 7 days of exposure. Our results demonstrate that both Termin-8 and Finio can be used as effective chemical mitigants against megaviruses such as EhV and ASFV and can be used as effective preventive or mitigation strategies to prevent the transmission of ASFV by reducing particle viability in contaminated feed, although additional research is warranted. Full article

Soil degradation exerts profound impacts on soil ecological functions, global food security, and human development, making the development of effective technologies to mitigate degradation a critical research focus. Microorganisms play a leading role in rehabilitating degraded land, improving soil hydraulic properties, and enhancing soil structural stability. Mosses contribute to soil particle fixation through their unique rhizoid structures; however, the mechanisms underlying their interactions in mixed inoculation remain unclear. Therefore, this study addresses soil and water loss caused by rainfall erosion in the cold black soil region. We conducted controlled laboratory experiments cultivating Bacillus subtilis and cold-adapted moss species, evaluating the erosion mitigation effects of different biological treatments under gradient slopes (3°, 6°, 9°) and rainfall intensities (70 mm h⁻¹, 120 mm h⁻¹), and elucidating their carbon-based structural reinforcement mechanism. The results indicated that compared to the control group, Treatment C significantly increased the mean weight diameter (MWD) and geometric mean diameter (GMD) of soil aggregates by 121.6% and 76.75%, respectively. In separate simulated rainfall events at 70 mm h⁻¹ and 120 mm h⁻¹, Treatment C reduced soil loss by 95.70% and 96.75% and decreased runoff by 38.31% and 67.21%, respectively. Crucially, the dissolved organic carbon (DOC) loss rate in Treatment C was only 21.98%, significantly lower than that in Treatment A (32.32%), Treatment B (22.22%), and the control group (51.07%)—representing a 59.41% reduction compared to the control. This demonstrates the following: (1) Bacillus subtilis enhances microbial metabolism, driving carbon conversion into stable pools, while mosses reduce carbon leaching via physical barriers, synergistically forming a dual “carbon protection–structural reinforcement” barrier. (2) The combined inoculation optimizes soil structure by increasing the proportion of large soil particles and enhancing aggregate stability, effectively suppressing soil loss even under extreme rainfall erosion. This study elucidates, for the first time, the biological pathway through which microbe–moss interactions achieve synergistic carbon sequestration and erosion resistance by regulating aggregate formation and pore water dynamics. It provides a scalable “carbon–structure”-optimized biotechnology system (co-inoculation of Bacillus subtilis and moss) for the ecological restoration of the cold black soil region. Full article

Avian influenza is an economically significant disease affecting poultry worldwide and is caused by influenza A viruses that can range from low to highly pathogenic strains. These viruses primarily target the respiratory, digestive, and nervous systems of birds, leading to severe outbreaks that threaten poultry production and pose zoonotic risks. The ectodomain of the avian influenza virus (AIV) matrix protein 2 (M2e), known for its high conservation across influenza strains, has emerged as a promising candidate for developing a universal influenza vaccine in a mouse model. However, the efficacy of such expression against poultry AIVs remains limited. The objective of this study was to evaluate the immunogenicity of nodavirus-like particles displaying the M2e proteins. In this study, three synthetic heterologous M2e genes originated from AIV strains H5N1, H9N2 and H5N2 were fused with the nodavirus capsid protein (NVC) of the giant freshwater prawn Macrobrachium rosenbergii (NVC-3xAvM2e) prior to immunogenicity characterisations in chickens. The expression vector pTRcHis-TARNA2 carrying the NVC-3xAvM2e gene cassette was introduced into E. coli TOP-10 cells. The recombinant proteins were purified, inoculated into one-week-old specific pathogen-free chickens subcutaneously and analysed. The recombinant protein NVC-3xAvM2e formed virus-like particles (VLPs) of approximately 25 nm in diameter when observed under a transmission electron microscope. Dynamic light scattering (DLS) analysis revealed that the VLPs have a polydispersity index (PDI) of 0.198. A direct ELISA upon animal experiments showed that M2e-specific antibodies were significantly increased in vaccinated chickens after the booster, with H5N1 M2e peptides having the highest mean absorbance value when compared with those of H9N2 and H5N2. A challenge study using low pathogenic AIV (LPAI) strain A/chicken/Malaysia/UPM994/2018 (H9N2) at 10^6.5 EID₅₀ showed significant viral load in the lung and cloaca, but not in the oropharyngeal of vaccinated animals when compared with the unvaccinated control group. Collectively, this study suggests that nodavirus-like particles displaying three heterologous M2e have the potential to provide protection against LPAI H9N2 in chickens, though the vaccine’s efficacy and cross-protection across different haemagglutinin (HA) subtypes should be further evaluated. Full article

Bovine alphaherpesvirus 1 (BoAHV-1) is a promising oncolytic virus that can infect the human lung carcinoma cell line A549. In an effort to adapt the virus to grow more rapidly in these cells through the serial passaging of viral progeny, we were unsuccessful. Here, we found that extracellular vesicles (EVs) secreted by BoAHV-1-infected A549 cells (referred to as EDVs) contain 59 viral proteins, including both viral structure proteins (such as gC and gD) and viral regulatory proteins (such as bICP4 and bICP22), as identified via a proteomic analysis. These EDVs can bind to and enter target cells, inhibit viral particles binding to cells, and stimulate the production of IFN-α and IFN-β in A549 cells. When EDVs are inoculated into rabbits via either the conjunctival sacs or intravenously, they can be readily detected in neurons within the trigeminal ganglia (TG), where they reduce viral replication and promote the transcription of IFN-γ. Furthermore, incorporation of the known anti-herpesvirus drug Acyclovir (ACY) into the EDVs leads to synergistically enhanced antiviral efficacy. Collectively, the EDVs exhibit antiviral effects by blocking viral binding to target cells and stimulating the innate immune response, thereby leading to the failure of the serial passaging of viral progeny in these cells, and these EDVs may serve as a promising vector for delivering drugs targeting TG tissues for antiviral purposes. Full article

This study evaluates the effects of solid-state fermentation inoculated with Aspergillus spp. on the nutritional profile of selected agro-industrial by-products (AIBPs: cowpea shell, groundnut shell, soybean hull, and maize shaft). These AIBPs were assessed as potential feedstuffs in weaner rabbit diets, which often exhibit digestive disorders when introduced to highly lignified feed ingredients. The AIBPs were milled to a particle size of 2 mm, sterilized, and subjected to fermentation with Aspergillus spp. under microaerophilic conditions at 28 ± 2 °C for 10 days. Samples (four replicates per treatment) were analyzed for chemical constituents (mineral and proximate composition, anti-nutritional factors, and fibre fractions) before and after fermentation. Digestible energy and digestibility coefficient of gross energy were calculated. Data were subjected to two-way analysis of variance (ANOVA). There was an increase (p < 0.05) in mineral profile, proximate composition, digestible energy, digestibility coefficient of gross energy, and dry matter, with a reduction (p < 0.05) in crude fibre, fibre fractions, and anti-nutritional factors. It was concluded that fermentation with Aspergillus spp. improved the nutritional value of the selected agro-industrial by-products. Therefore, fermented materials possess a better nutritional profile to be used in feeding programs for weaner rabbits. This will ensure sustainable animal production and add value to agricultural waste, which would otherwise constitute an environmental nuisance. Full article

Parainfluenza viruses are a common cause of respiratory illness in many species. In this study, experimental, alphavirus-derived RNA particle vaccines either with or without adjuvant were evaluated against porcine parainfluenza virus 1 (PPIV1) challenge and compared to live virus exposure. Groups of ten, three-week-old pigs were vaccinated intramuscularly with an adjuvanted RNA particle (RPAdj/C) or non-adjuvanted RP (RP/C) or administered an intranasal live exposure (LE/C) dose of PPIV1 at 0- and 21-days post vaccination (DPV) followed by challenge with PPIV1 at 40 DPV. In addition, two groups were included as non-vaccinated, non-challenged (NV/NC) and non-vaccinated, challenged (NV/C) controls. Intranasal virus exposure and RP vaccination, regardless of adjuvant, reduced PPIV1 shedding in nasal swabs by 5 days post inoculation (DPI). All vaccinated or exposed pigs seroconverted as shown by enzyme-linked immunosorbent assay and serum virus neutralization. The antibody isotype detected in bronchoalveolar lavage fluid (BALF) LE/C was predominantly IgA while RP vaccination induced an IgG response. Reduced PPIV1 antigen was observed in the LE/C, RP/C and RPAdj/C groups in lung, trachea, or nasal turbinate epithelium. Additionally, the RPAdj vaccine significantly reduced nasal shedding compared to NV/C pigs although not as much as LE/C pigs. These results suggest vaccination could mitigate PPIV1 infection in commercial systems. Full article

In order to investigate the causes of swine diarrhea in Yunnan Province, this study was conducted to detect and monitor diarrhea viruses through regular sampling and reverse transcription polymerase chain reaction (RT-PCR). In October 2023, porcine sapelovirus (PSV) was detected in fecal specimens collected from diarrheal pigs in Honghe City, and three strains of PSV were successfully isolated by inoculating them into PK-15 cells; electron microscopy revealed virus particles with diameters of ~32 nm. Next-generation sequencing (NGS) revealed that the PSV isolate genomes ranged from 7480 to 7515 nucleotides in length. Homology analyses indicated that ML-15 and ML-16 showed the highest nucleotide and amino acid identities with the Asian PSV strains, ML-19 showed the highest sequence identities with the Zambia PSV strains, and the VP1 to VP4 genes of the three PSV isolates were in the hypervariable region. Phylogenetic analyses showed that the three PSVs isolated in this study all clustered together with Chinese PSV strains; furthermore, recombination analyses indicated that PSV-ML-19 might be a recombined strain and may have emerged through genetic recombination between the major putative parent strain PSV-21-V and the minor putative parent GER L00798-K11 14-02. This was the first reported instance of the isolation and phylogenetic analyses of the PSV strains in Yunnan Province, which enriched the understanding of Chinese PSV strains and indicated the need to prevent and control PSV; the mutation of the VP1 and 3D genes may also provide an important reference for the development of PSV vaccines. Full article

The aim of this study was to develop a novel functional ingredient—goat’s skim milk enriched with Agrocybe aegerita (V. Brig.) Vizzini mushroom extract (ME/M)—using Central Composite Design (CCD). The optimized ME/M ingredient was evaluated for its physico-chemical, techno-functional, biological, and antimicrobial properties. Physico-chemical properties were analyzed using Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) spectroscopy, Scanning Electron Microscopy (SEM), and Dynamic Light Scattering (DLS). The ingredient exhibited a polymodal particle size distribution and contained glucans, along with a newly formed polypeptide resulting from the selective cleavage of goat milk proteins. A 0.1% ME/M solution demonstrated good emulsifying and foaming properties. Additionally, ME/M showed strong antiproliferative effects on human cancer cell lines, particularly Caco-2 (colorectal) and MCF7 (breast) cancer cells. The ingredient also promoted HaCaT cell growth without cytotoxic effects, suggesting its safety and potential wound-healing properties. Furthermore, the addition of ME/M to HaCaT cells inoculated with Staphylococcus aureus resulted in reduced IL-6 levels compared to the control (without ME/M), indicating a dose-dependent anti-inflammatory effect. The optimized ME/M ingredient also exhibited antibacterial, antifungal, anticandidal, and antibiofilm activity in one-fourth of MIC. These findings suggest that the formulated ME/M ingredient has strong potential for use in the development of functional foods offering both desirable techno-functional properties and bioactive benefits. Full article

Plastic pollution is a global emerging concern, but in the Arctic Ocean, the role of the plastisphere as a potential carrier of pathogens and antibiotic-resistant bacteria is unknown yet. An initial assessment of the spread of these target bacteria through their colonization of plastic particles, attributed to the micro-sized fraction (less than 5 mm, named microplastics, MPs), was carried out across the 75° N transect (Greenland Sea). To fill the knowledge gaps regarding the bacterial community associated withmicroplastics (MPs)—belonging to the so-called “plastisphere”—and the potential risks related to their spread, our study focused on the abundance and taxonomic composition of the plastisphere, including potential pathogenic and antibiotic-resistant bacteria, using a culture-dependent approach. MPs particles were collected through a Manta net, and decimal dilutions were cultured on Marine agar plates to estimate the abundance of culturable heterotrophic bacteria. For the search of target pathogenic bacterial species (Escherichia coli, Enterococcus spp., Salmonella spp., potentially pathogenic Vibrio spp., and Staphylococcus aureus), small volumes were inoculated into selective culture media using aspread plate directly or after enrichment. Screening of the antibiotic susceptibility profiles of the bacterial isolates was performed to assess the presence of antibiotic-resistant bacteria. The culturable heterotrophic bacterial community was dominated by members of the phyla Gammaproteobacteria and Actinobacteria, with bacterial isolates assigned to the genera Psychrobacter, Pseudoalteromonas, Shewanella, and Arthrobacter. Selective enrichments resulted in the detection of target bacterial pathogens, mostly identified as potentially pathogenic Vibrios spp., in the examined samples. The antibiotic susceptibility profiles pointed out that multiple antibiotic-resistant bacteria were also isolated, suggesting the need to shed light on the potential risks to human and animal health deriving from the plastisphere in remote cold regions as well. Full article

This work determined the resistance of paint formulations containing TiO₂ particles to fungal growth. Siloxane, acrylic and silicone paints were placed outdoors, and the fungal species growing thereon were recorded after 3, 6 and 9 months. In addition, three paint types containing TiO₂ with/without biocide were inoculated with fungal spores and irradiated using UV. Acrylic paints were also doped with different metals and were inoculated and incubated under fluorescent light. Following outdoor incubation, the silicone paint was the least colonised by different fungal species. The species most recovered from the surfaces were Aspergillus spp. and Penicillium spp. Following UV irradiation on different paints containing biocide and/or a photocatalyst, no fungal growth was demonstrated on some of the paint combinations. When the paint samples were doped with different metals and incubated using light, the sample most efficient at preventing fungal growth contained lanthanum (0.004%). The paint samples containing praseodymium (light:1.72) facilitated the densest fungal colonies. Most of the surfaces demonstrated heterogeneous coverage by the fungi. The most clustered fungal colonisation was on surfaces incubated in the light. This work demonstrated that fungal colonisation on paints changed over time and that the antimicrobial efficacy of TiO₂ was affected by the chemical composition, biocide and doping of the paint. Full article

The Spodoptera frugiperda Sf9 insect cell line is used in the baculovirus expression vector system for the development of various viral vaccines and some gene therapy products. Early studies indicated that Sf9 cells produced a reverse transcriptase (RT) activity that was detected using a sensitive PCR-enhanced reverse transcriptase (PERT) assay. Since RT is generally associated with retrovirus particles, we undertook the investigation of the physical properties and infectious nature of the extracellular RT activity that was constitutively expressed from Sf9 cells or induced after the chemical treatment of the cells with drugs known to activate endogenous retroviruses. A density gradient analysis indicated that the peak RT activity corresponded to a low buoyant density of about 1.08 g/mL. Ultracentrifugation and size filtration of cell-free Sf9 supernatant indicated that different particle sizes were associated with the RT activity. This was confirmed by transmission electron microscopy and cryoEM, which revealed a diversity in particle size and type, including viral-like and extracellular vesicles. The treatment of Sf9 cells with 5-iodo-2′-deoxyuridine (IUdR) induced a 33-fold higher RT activity with a similar low buoyant density compared to untreated cells. Infectivity studies using various target cells (human A204, A549, MRC-5, and Raji, and African green monkey Vero cells) inoculated with cell-free supernatant from untreated and IUdR-treated Sf9 cells showed the absence of a replicating retrovirus by PERT-testing of cell-free supernatant during the 30 day-culturing period. Additionally, there was no evidence of virus entry by whole genome analysis of inoculated MRC-5 cells using high-throughput sequencing. This is the first study to identify extracellular retroviral-like particles in Spodoptera. Full article

The antifungal efficacy of coffee residue extract (CRE) and chitosan nanoparticles (CNPs) alone and in combination (CNPs-CRE) against the growth of Rhizopus stolonifer was assessed. Two nanostructured edible coatings (ECs), one consisting of chitosan nanoparticles (CCNP) and another consisting of coffee residue extract encapsulated in chitosan nanoparticles (CCNP-CRE), were elaborated, characterized, and applied on inoculated Naples tomatoes to evaluate their fungicidal activity and their effect on fruit quality. The tests consisted of evaluating physicochemical variables in tomato previously sprayed with CCNP and CCNP-CRE for 30 days preharvest and 14 days at 10 °C postharvest. CNPs with a particle size of 2.4 ± 0.24 nm with a Z potential of −1.62 mV were observed, while CNPs-CRE showed a size of 3.9 ± 0.55 nm with a Z potential of −0.89 mV. The FTIR spectrum showed the integration of CRE into the CCNP-CRE. A synergistic effect between CNPs and 1% CRE was observed, obtaining the greatest inhibition of mycelial growth (43%). In the preharvest trials, both ECs showed differences with respect to the control in the variables of color, total carotenoids and ethylene. In the postharvest test, they showed differences in color and CO₂ production. The severity of the infection decreased by 33% in tomatoes with coatings. The ECs evaluated represent a emergent technological advancement in the conservation of the tomato. Full article

The immobilisation of essential oil components (EOCs) on food-grade supports is a promising strategy for preserving liquid foods without the drawbacks of direct EOC addition such as poor solubility, high volatility, and sensory alterations. This study presents a novel method for covalently immobilising EOCs, specifically thymol and carvacrol, on SiO₂ particles (5–15 µm) using the Mannich reaction. This approach simplifies conventional covalent immobilisation techniques by reducing the steps and reagents while maintaining antimicrobial efficacy and preventing compound migration. The antimicrobial effectiveness of the EOC–SiO₂ system, applied as an additive, was tested against foodborne pathogens (Escherichia coli, Salmonella enterica, Staphylococcus aureus, and Listeria monocytogenes) inoculated into phosphate buffer solution and fresh apple juice. The results showed high antimicrobial activity, with inactivation exceeding 4-log reductions, depending on the EOC type, target microorganism, and medium. Moreover, the addition of functionalised particles did not affect the juice organoleptic properties. This study demonstrates that the Mannich reaction is an effective method for developing antimicrobial systems based on the covalent immobilisation of EOCs on silica particles, and offers a practical solution for food preservation without compromising food quality. Full article