Search Results (3,538)

Reproductive failure in cattle production is a global concern and is influenced by various factors, including genetic alterations. This study explored the relationship between an X-linked single-nucleotide variant (NC_037357.1: g.87298881A>G, rs135720414) in the upstream of the bovine forkhead box P3 (FOXP3) gene and infertility. To this end, we examined the genotypes of the variant in old Asian cattle breeds, including 48 Bali and 5 Jaliteng cattle, and 20 water buffaloes, which have recently shown subclinical signs of infertility and repeated breeding problems among populations in Indonesia. We also examined the genotypes in 69 parous and 39 non-parous Holstein Friesian (HF) cows and investigated the relationship between the genotypes and serum concentration of anti-Müllerian hormone (AMH). The G allele frequency was markedly high in Bali (0.944) and Jaliteng cattle (0.714), and water buffaloes (1), suggesting that the G allele may be originally a wild-type variant in old Asian cattle and buffaloes. In HF cows, the G allele frequency was moderately high, and the AMH concentration was significantly lower (p < 0.05) in parous cows carrying the G allele (A/G and G/G genotypes) than in parous cows with the A/A genotype. In contrast, there were no significant differences in AMH concentrations among the three genotypes of non-parous HF cows. This suggests that both G allele and aging are associated with infertility in HF cows. In conclusion, the G allele of the FOXP3 gene variant may potentially be associated with infertility in different bovine breeds and species. Therefore, special attention should be paid to this variant, and infertility in bovine herds may be improved by selection and/or introduction of the A allele. Full article

Acmella radicans, commonly known as the “toothache plant,” is traditionally attributed with medicinal properties, although few studies have validated its biological effects. In the present study, a chemical analysis of the wild plant was performed using gas chromatography–mass spectrometry (GC-MS). In addition, the antioxidant, anti-inflammatory, and antibacterial potential of ethanolic extracts from the roots (RE) and aerial parts (AE), as well as their respective fractions, was evaluated. The dichloromethane fractions of the aerial parts (DFAE) and root extracts (DFRE) at a concentration of 25 μg/mL demonstrated the highest inhibition of nitric oxide (NO) production, reducing levels to 22.2 ± 1.9 and 22.2 ± 2.9 μM, respectively. Moreover, these fractions exhibited a notable inhibition of TNF-α production, lowering its concentration to 22.6 ± 3.3 pg/mL (DFAE) and 24.8 ± 5.3 pg/mL (DFRE) at 25 µg/mL. GC-MS chemical profiling revealed the presence of alkamides such as N-isobutyl-2E,6Z,8E-decatrienamide, N-(2-methylbutyl)-2E,6Z,8E-decatrienamide, and N-(2-phenylethyl)-2E,4Z-octadienamide in both root and aerial part extracts. The dichloromethane fractions showed a higher abundance of alkamides compared to the hexane fractions, suggesting that these compounds may be at least partially responsible for the observed anti-inflammatory activity. Additionally, AE showed moderate activity against S. typhimurium and low activity against other bacteria, while RE was especially effective against a resistant strain of S. aureus, indicating an MIC of 31.25 μg/mL, likely due to its high content of alkamides, particularly spilanthol. Several fractions also inhibited bacteria such as P. aeruginosa, S. aureus, and E. coli, possibly because of the presence of alkamides and compounds like β-amyrin. Full article

The utilization of organic fertilizers for the cultivation of wild edible and medicinal plants offers agronomic and ecological benefits, given their suitability to low-input and sustainable production systems. Under such conditions, these species may also benefit from targeted foliar applications of micronutrients to enhance their nutritional quality. This study examined the effects of a vinasse-based organic fertilizer and conventional fertilization regime, in combination with foliar applications of iron (Fe) and zinc (Zn), on the biomass, leaf photochemistry, and plant stress-related responses of Sideritis cypria and Origanum dubium. In S. cypria, organic fertilization resulted in a similar yield compared to conventional fertilization, while O. dubium showed a significant decrease in yield when using organic fertilizers. The impact of spraying with Zn on S. cypria dry matter content was related to the availability of nutrients, particularly nitrogen, while in O. dubium Zn spraying induced a decrease in dry matter. The total phenols content and antioxidant activity of S. cypria were elevated by conventional fertilization and foliar application of Fe, while the combination of organic fertilization and foliar application of Fe and Zn reduced lipid peroxidation. In O. dubium, foliar application of Fe and Zn led to a reduction in total phenols content, antioxidant capacity, and hydrogen peroxide content under adequate nutrition. In general, foliar spraying with Zn tended to improve water use efficiency under specific fertilization practices on both species, while the positive effect of conventional fertilization on nutrient use efficiency still requires further validation. Ultimately, the efficiency of organic fertilization was related to the examined species, inducing variations in leaf chlorophyll content. In addition, foliar application of Fe and Zn affected the antioxidant capacity and mineral content of the examined species. Thus, appropriate fertilization management is vital to fully realize the specific benefits of foliar micronutrient addition. Full article

Leaf morphology significantly impacts rice (Oryza sativa L.) plant architecture and yield. Here, we identified and characterized a novel narrow-leaf mutant, nal25, derived from indica rice cultivar ‘Huazhan’ using EMS mutagenesis. Phenotypic analyses revealed that nal25 exhibited significantly narrower leaves, reduced plant height, increased tiller number, and notably decreased grain size, seed setting rate, and thousand-grain weight compared to the wild type. Genetic analyses demonstrated that the narrow-leaf phenotype is controlled by a single recessive nuclear gene. Through precise localization analysis, the NAL25 gene was located within a region of approximately 103 kb on the long arm of rice chromosome 7. The sequencing results showed that the mutant nal25 had a T to C mutation at position 173 of the heat-shock protein gene LOC_Os07g09450 encoding the DnaJ domain in this interval, resulting in a change in amino acid 58 from leucine to proline. The qRT-PCR results showed that the expression level of NAL25 gene decreased in the mutant. The nal25 mutant obtained in this study exhibits stable mutant phenotypes, including dwarfism and excessive tillering, traits typically unfavorable for rice production. Nevertheless, it serves as valuable genetic material for forward genetics approaches to identify yield-related genes regulating leaf morphology and culm height. Thus, research on the nal25 mutant advances the development of rice varieties with ideal plant architecture, thereby stabilizing yield increases and safeguarding global food security. Full article

This study highlights a new avenue to improve polyhydroxybutyrate (PHB) productivity by optimizing genes related to arginine catabolism, which influences nitrogen metabolism in cyanobacteria based on the carbon/nitrogen metabolism balance. In the Synechocystis sp. PCC 6803 wild type (WT) and its adc1 mutant (Δadc1), the native putA gene, responsible for the oxidation of proline to glutamate, was overexpressed to create the OXPutA and OXPutA/Δadc1 strains, respectively. PHB accumulation was considerably higher in OXPutA and OXPutA/Δadc1 under the nitrogen-deprived condition than in strains that overexpressed the proC gene, involved in proline synthesis. The increased transcript level of glgX, associated with glycogen degradation, confirmed that glycogen served as the primary carbon source for PHB synthesis under nitrogen stress without any carbon source addition. Furthermore, proline and glutamate level changes helped cells deal with nitrogen stress and considerably improve intracellular carbon/nitrogen metabolism. As indicated by elevated levels of proA and argD transcripts as well as chlorophyll a accumulation, this impact was most noticeable in strains that overexpressed putA, which was crucial for the synthesis of glutamate, a precursor for important metabolic pathways that respond to nitrogen stress. Therefore, our metabolic model presents PHB-producing strains as promising candidates for biomaterial biotechnology applications in medical and agricultural fields. Full article

Sebum secreted by sebaceous glands mixes with sweat to form a protective film that aids in maintaining skin health. Reduced sebum production compromises such barrier functions, potentially leading to severe itchiness and inflammation. Therefore, incorporating moisturizers with ingredients promoting sebum secretion is desirable. Wild watermelon possesses moisturizing and antioxidant properties, and its extracts are utilized in skin cosmetics and supplements. This study investigates whether seed watermelon (Citrullus mucosospermus (Fursa))—a species closely related to wild watermelon—influences sebum synthesis and can serve as a skin cosmetic raw ingredient. Several bioactive compounds—including coniferyl alcohol, coniferin, and p-coumaryl alcohol—were identified in the active third fraction of the fruit extract. Subsequently, SZ95 sebocytes stimulated with linoleic acid were stained using Oil Red O to detect lipogenesis facilitated by the identified bioactive compounds. Coniferyl alcohol promoted linoleic acid-stimulated lipogenesis by approximately 2.2-fold at a concentration of 300 µM. Lipidomic analyses confirmed an increase in total lipid content following coniferyl alcohol treatment, with notable increases in cholesterol ester, cardiolipin, and simple lipid content. Overall, these findings suggest that seed watermelon contains compounds that do influence sebum synthesis. Consequently, skin cosmetics containing seed watermelon fruit extracts with linoleic acid may benefit individuals with dry skin. Full article

Rhaponticum carthamoides (Willd.) Iljin. (Leuzea carthamoides, Maral root), a medicinally valuable species listed in the Red Book of Kazakhstan, is known for its rich phytochemical profile. However, limited data exist on its microclonal propagation. This study aimed to optimize in vitro and medium-term storage conditions using biotechnological methods. Mature seeds collected from natural populations in the Kazakhstani Altai were germinated, and tissues from the seedlings were used as explants. Sterile shoots were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg L⁻¹ −6-benzylaminopurine and 3.0 mg L⁻¹ kinetin. For shoot induction, MS medium supplemented with 0.5 mg L⁻¹ meta-Topolin and using stem apices as explants yielded optimal results. Medium-term storage with chlorocholine chloride at 0.1–0.4 g/L effectively preserved regenerative capacity for further rooting. After 12 months of storage, plantlets were transferred to half-strength MS medium with 3.0 g/L activated carbon and at 2.0 or 5.0 mg L⁻¹ indole-3-butyric acid for rooting. Regenerated plants were successfully acclimatized ex vitro. The 20-hydroxyecdysone content in field-grown plants post-storage reached 9.24 mg/mL, 2.4-fold higher than in wild plants. Inter simple sequence repeat analysis confirmed genetic stability. Our optimized protocol ensures high-yield metabolite production and genetic fidelity, enabling in vitro conservation, nursery-scale cultivation, and the restoration of R. carthamoides natural populations. Full article

Background: The Citrobacter genus harbors class C (AmpC) and class A β-lactamases. Citrobacter freundii produces an inducible AmpC β-lactamase controlled by the LysR-type transcriptional regulator AmpR and cytosolic amidase AmpD. Citrobacter sedlakii produces the class A β-lactamase Sed-1, whose expression is believed to be regulated by the transcriptional regulator SedR and AmpD. Objectives:C. sedlakii NR2807, isolated in Japan, is resistant to third-generation cephalosporins and displays extended-spectrum β-lactamase characteristics. Here, we sought to understand the mechanism for successful resistance to third-generation cephalosporins by investigating the regulators controlling Sed-1 production. Methods: Plasmids containing bla_Sed-1 and sedR (pCR2807) or truncated sedR (pCR2807ΔSedR) were constructed and introduced into Escherichia coli. Antibiotic-resistant mutants of NR2807 were obtained, and enzyme kinetics were assessed. Results: The AmpD mutant (pCR2807/ML4953) showed an 8-fold increase in cefotaxime MIC and an 8.46-fold increase in Sed-1 activity compared to the wild-type (pCR2807/ML4947). However, induction of pCR2807/ML4947 also led to a 1.32-fold higher Sed-1 activity, indicating semi-inducibility. Deletion of sedR (pCR2807ΔSedR/ML4947) led to a 4-fold decrease in cefotaxime MIC and 1.93-fold lower Sed-1 activity, confirming SedR as an activator. While wild-type C. sedlakii ATCC51115 is susceptible to third-generation cephalosporins, the AmpD mutation in NR2807 led to Sed-1 overproduction and resistance to this class of antibiotics. Finally, mutagenesis revealed that amino acid substitution in Sed-1 conferred resistance to ceftazidime and extended-spectrum β-lactamase characteristics. Conclusions: Sed-1 producers, though usually susceptible to third-generation cephalosporins, may develop extended-spectrum β-lactamase traits due to AmpD or Sed-1 mutations, thereby requiring careful monitoring. Full article

The present work evaluates the influence of two prebiotic formulations—P1 (ColonX) and P4 (a product containing AnXietate extract), at concentrations of 3 and 6 mg/L—on scopolamine (SCOP, 100 μM)-induced cognitive dysfunction and anxiety-related behaviors in adult zebrafish (Danio rerio). To assess behavioral alterations, wild-type fish were subjected to the novel tank assay (NTT), Y-maze study, and novel object recognition test protocol (NOR). The formulations were examined for potential anti-inflammatory activity and cytotoxicity. In parallel, in vitro assays were performed to evaluate cytotoxic and anti-inflammatory effects. The results indicate that both prebiotic formulations effectively mitigated SCOP-induced behavioral impairments and improved cognitive performance in zebrafish. Furthermore, the prebiotic formulation P4 showed significant anti-inflammatory activity without inducing cytotoxicity. The study was conducted following ethical guidelines, ensuring scientific rigor and integrity. These findings highlight the therapeutic potential of prebiotics in alleviating anxiety and cognitive deficits, with promising implications for the management of neuropsychiatric disorders. Full article

Background/Objectives: The Type IV pilus assembly system in Acinetobacter baumannii is a major determinant of its pathogenicity, playing a role in surface-associated functions via the biogenesis of Type IV pili (T4P). Tazobactam (TAZ) is a well-characterized β-lactamase inhibitor, primarily used in combination with β-lactam antibiotics such as piperacillin (PIP) to counteract bacterial resistance mechanisms. While A. baumannii resistance to β-lactam antibiotics has been well studied, the influence of T4P on its susceptibility to TAZ remains largely unexplored. For this reason, we investigated how multidrug-resistant A. baumannii 5075 (AB5075) responds to TAZ by assessing the roles of pilA, pilB, and pilD in bacterial growth and biofilm formation under direct TAZ exposure, with a focus on phenotypic characterization rather than molecular mechanisms. Methods: Bacterial growth kinetics were quantified by measuring the optical densities of cell suspensions and the colony forming units per volume (CFUs/mL) at different time intervals. Time-kill assays and microtiter dish biofilm formation assays were used to evaluate how effectively TAZ can inhibit growth and biofilm formation, respectively. Results: Time–kill assays confirmed that 32 µg/mL of TAZ inhibited growth in both wild-type (WT) and mutant strains, with the pilD mutant showing initial resistance before eventual inhibition. Biofilm assays showed that the pilA mutant had the highest biofilm formation at 8 h, surpassing the WT strain. A prolonged 32 µg/mL of TAZ exposure (24–36 h) significantly reduced biofilm production across all strains, with inhibition rates reaching 89% for the WT, 82% for the pilA mutant, 91% for the pilB mutant, and 86% for the pilD mutant. Conclusion: These findings deepen our understanding of the strain-specific roles of T4P components in growth and biofilm regulation in AB5075, and highlight the potential of TAZ as a therapeutic strategy against biofilm-associated infections. Full article

Cell culture-based influenza vaccines exhibit comparable safety and immunogenicity to traditional egg-based vaccines. However, improving viral yield remains a key challenge in optimizing cell culture-based production systems. Madin–Darby canine kidney (MDCK) cells, the predominant cell line for influenza vaccine production, inherently activate interferon (IFN)-mediated antiviral defenses that restrict viral replication. To overcome this limitation, we employed CRISPR/Cas9 gene-editing technology to generate an IFN alpha/beta receptor subunit 1 (IFNAR1)-knockout (KO) adherent MDCK cell line. Viral titer analysis demonstrated significant enhancements in the yield of multiple vaccine strains (H1N1, H3N2, and type B) in IFNAR1-KO cells compared to wild-type (WT) cells. Transcriptomic profiling revealed marked downregulation of key interferon-stimulated genes (ISGs)—including OAS, MX2, and ISG15—within the IFNAR1-KO cells, indicating a persistent suppression of antiviral responses that established a more permissive microenvironment for influenza virus replication. Collectively, the engineered IFNAR1-KO cell line provides a valuable tool for influenza virus research and a promising strategy for optimizing large-scale MDCK cell cultures to enhance vaccine production efficiency. Full article

Plastic waste, particularly poly(ethylene terephthalate) (PET), negatively impacts the environment and human health. Biotechnology could become an alternative to managing PET waste if enzymes ensure the recovery of terephthalic acid with efficiencies comparable to those of chemical treatments. Recent research has highlighted the potential of fungal cutinases, such as wild-type ANCUT1 (ANCUT1^wt) from Aspergillus nidulans, in achieving PET depolymerization. Fungal cutinases’ structures differ from those of bacterial cutinases, while their PET depolymerization mechanism has not been well studied. Here, a reliable model of the ANCUT1^wt was obtained using AlphaFold 2.0. Computational chemistry revealed potential cation-binding sites, which had not been described regarding enzymatic activation in fungal cutinases. Moreover, it allowed the prediction of residues with the ability to interact with a PET trimer that were mutation candidates to engineer the substrate binding cleft, seeking enhancements of PET hydrolysis. Enzyme kinetics revealed that both ANCUT1^wt and ANCUT1^N73V/L171Q (DM) were activated by MgCl₂, increasing the dissociation constant of the substrate and maximal reaction rate. We found that in the presence of MgCl₂, DM hydrolyzed different PET samples and released 9.1-fold more products than ANCUT1^wt. Scanning Electron Microscopy revealed a different hydrolysis mode of these enzymes, influenced by the polymer’s crystallinity and structure. Full article

Helminth parasites of wild animals represent a major threat to the health of these animals, leading to significant losses in performance, health, and zoonotic implications. In some zoos, anthelmintics have traditionally been used to control these parasites, many of which are also zoonotic. Other actions, such as the removal of organic waste, have also been adopted. Few or no control measures are applied to free-ranging wild animals. Helminthophagous fungi are a promising biological alternative. When animals ingest fungal spores, they are excreted in their feces, where they trap and destroy helminth larvae and eggs, preventing and reducing the parasite load in the environment. Another alternative is to administer fungi by spraying them directly into the environment. This review aims to examine the use of helminthophagous fungi in the control of helminthiases in wild animals, highlighting their potential to minimize dependence on chemical treatments and promote sustainable animal breeding and production. There are many challenges to making this viable, such as environmental variability, stability of formulations, and acceptance of this new technology. These fungi have been shown to reduce parasite burdens in wild animals by up to 75% and can be administered through the animals’ feeding troughs. To date, evidence shows that helminthophagous fungi can reliably curb environmental parasite loads for extended periods, offering a sustainable alternative to repeated anthelmintic dosing. Their use has been linked to tangible gains in body condition, weight, and overall welfare in various captive and free-ranging wildlife species. Full article

Diploid Vaccinium fuscatum is a wild blueberry species with a low chilling requirement, an evergreen growth habit, and soil adaptability to southeast US growing regions. Regardless of its potential to improve the abiotic and biotic resilience of cultivated blueberries, this species has rarely been used for blueberry breeding. One hurdle is the ploidy barrier between diploid V. fuscatum and tetraploid cultivated highbush blueberries. To overcome the ploidy barrier, vegetative shoots micro-propagated from one genotype of V. fuscatum, selected because it grew vigorously in vitro and two southern highbush cultivars, ‘Emerald’ and ‘Rebel,’ were treated with colchicine. While shoot regeneration was severely repressed in ‘Emerald’ and ‘Rebel,’ shoot production from the V. fuscatum clone was not compromised at either 500 µM or 5000 µM colchicine concentrations. Due to the high number of shoots produced in vitro via the V. fuscatum clone shoots of this clone that had an enlarged stem diameter in vitro were subjected to flow cytometer analysis to screen for induced polyploidy. Sixteen synthetic tetraploid V. fuscatum, one synthetic octoploid ‘Emerald,’ and three synthetic octoploid ‘Rebel’ were identified. Growth rates of the polyploid-induced mutants were reduced compared to their respective wildtype controls. The leaf width and length of synthetic tetraploid V. fuscatum and synthetic octoploid ‘Emerald’ was increased compared to the wildtypes, whereas the leaf width and length of synthetic octoploid ‘Rebel’ were reduced compared to the wildtype controls. Significant increases in stem thickness and stomata guard cell length were found in the polyploidy-induced mutant lines compared to the wildtypes. In the meantime, stomata density was reduced in the mutant lines. These morphological changes may improve drought tolerance and photosynthesis in these mutant lines. Synthetic tetraploid V. fuscatum can be used for interspecific hybridization with highbush blueberries to expand the genetic base of cultivated blueberries. Full article

Antimicrobial resistance (AMR) is a growing global health threat, with wild birds increasingly recognized as potential reservoirs of resistant pathogens and as sentinels of environmental AMR. This study investigated the occurrence and AMR profiles of Gram-negative bacteria isolated from wild birds that died at the Wildlife Rescue Center in Vanzago, Lombardy, in 2024. Cloacal swabs were collected from 112 birds representing various ecological categories. A total of 157 Gram-negative bacteria were isolated and identified, including clinically relevant genera and species, such as Escherichia coli, Klebsiella pneumoniae, Enterobacter spp., Salmonella spp., Pseudomonas aeruginosa, and Acinetobacter baumannii. Antimicrobial susceptibility testing revealed resistance to first-line and critically important antimicrobials, including those exclusively authorized for human use. Notably, a phenotype compatible with Extended-Spectrum Beta-Lactamase (ESBL) production was detected in four out of ten (40%) K. pneumoniae isolates. In addition, 20 out of the 157 (12.7%) isolated bacteria phenotypically exhibited a resistance profile indicative of AmpC beta-lactamase (AmpC) production, including Enterobacter spp. and P. aeruginosa. Resistance patterns were particularly interesting in birds with carnivorous, scavenging, or migratory-associated behaviors. These findings highlight the role of wild birds in the ecology and dissemination of antimicrobial-resistant bacteria (ARB) and highlight the need for wildlife-based AMR monitoring programs as part of a One Health approach. Full article