Introduction Spontaneous abortion causes approximately 50,000 maternal deaths annually, primarily due to microbial infection and hemorrhage, accounting for 8–13% of global maternal deaths. This study aimed to investigate the association between microbial infections, bacterial diversity, and patterns related to spontaneous abortion in two groups of women. Methods A total of 156 bacterial isolates were recovered from 48 patient samples, while 67 isolates were obtained from 48 healthy controls. All samples were cultured under aerobic and anaerobic conditions on enrichment and selective media, including Blood agar, MacConkey agar, and Chocolate agar. Results Gram-positive bacteria predominated among total isolates (53.84%) compared to Gram-negative bacteria (46.15%). The primary Gram-positive pathogens were S. aureus (29.16%), L. acidophilus (27.08%), Peptostreptococcus spp. (22.91%), and C. perfringens 20.83%. Among Gram-negative isolates, E. coli was most prevalent (47.91%), followed by P. aeruginosa (18.75%) and K. pneumonia (16.6%). In the control group, L. acidophilus (43.75%) and E. coli (18.75%) were most common. Diversity analyses showed no significant difference in α-diversity, but β-diversity revealed clear distinctions between groups. Linear Discriminant Analysis identified specific biomarkers differentiating the microbial compositions of the two cohorts. Discussion These findings indicate a Gram-positive predominance, with Staphylococcus aureus and E. coli identified as the primary pathogenic drivers in the study groups. Although species richness remained consistent across cohorts, β- diversity and Linear Discriminant Analysis biomarkers confirmed distinct microbial shifts and bacterial signatures between the examined groups. Conclusion L. acidophilus appears to help maintain vaginal health and reduce the risk of spontaneous abortion, as β- diversity and biomarker analyses revealed a shift from beneficial to pathogenic bacteria in women with spontaneous abortion.
Introduction The growing application of biosides post-COVID-19 has heightened concern over the development and spread of antiseptic resistance determinants, especially qac genes in different types of bacteria. This study aimed to evaluate antiseptic resistance determinants in clinical P. aeruginosa isolates and to determine their response to antimicrobial resistance and biofilm-forming ability. Methods Fifty clinical P. aeruginosa isolates were evaluated for antimicrobial susceptibility using the Kirby-Bauer disk diffusion method, Biofilm formation using the microtiter plate crystal violet assay, and biocide tolerance by broth microdilution. PCR was performed to identify qacE and qacEΔ1 genes in selected highly resistant isolates. Statistical analysis was applied to identify the correlation among variables. Results The findings showed a high rate of MDR (64.0%) and XDR (6.0%). Biofilm assays showed strong (38.0%), moderate (30.0%), and weak (30.0%) production. Furthermore, strong /moderate biofilm formation was significantly associated with MDR/XDR phenotypes ( p < 0.0001 ). Reduced antiseptic sensitivity was detected in (60.0%) of isolates and strongly correlated with both antimicrobial resistance status and biofilm strength ( p < 0.0001 ). Molecular profiling of selected resistance isolates showed a universal presence of the qacEΔ1 gene (100%) and high prevalence of the qacE gene (73.3%). Discussion The significant correlation between qac genes and strong biofilm ability demonstrates a key mechanism of antibiotic and biocide tolerance in P. aeruginosa. Conclusion Qac genes are highly prevalent in clinical P. aeruginosa isolates and strongly correlated with multidrug resistance and biofilm-forming ability. Ongoing molecular monitoring of the biocide resistance genes in clinical setups is vital to prevent the propagation of exceptionally stubborn strains in clinical environments.
PPCPs have emerged as a significant class of environmental contaminants due to their extensive global consumption and persistence in natural ecosystems. Conventional wastewater treatment plants are often unable to completely remove these compounds, resulting in their continued release into aquatic and terrestrial environments. The bioaccumulation and long-term persistence of PPCPs raise serious concerns regarding potential ecological risks, disruption of microbial communities, and adverse impacts on human health through food chains and water reuse. This review provides a comprehensive synthesis of current advances in the biodegradation of PPCPs, with a particular focus on microbial degradation pathways. The roles of diverse microbial groups, including bacteria, fungi, and algae, as well as specific enzymatic systems, in the transformation and mineralization of PPCPs are critically examined. Special emphasis is placed on ligninolytic fungi, whose extracellular oxidative enzymes, such as laccases, peroxidases, and oxidoreductases, show high potential for degrading structurally complex PPCPs. Furthermore, the application of genetically engineered microorganisms is discussed as a promising strategy to enhance biodegradation efficiency by tailoring metabolic pathways. The review also evaluates environmental and physicochemical factors such as pH, temperature, redox potential, and the molecular properties of PPCPs that influence degradation outcomes. Beyond laboratory findings, advanced biotechnological innovations and decentralized on-site treatment technologies for soil and water systems are highlighted as practical approaches for sustainable remediation. By consolidating recent research findings, this work not only identifies existing challenges and knowledge gaps but also outlines future directions and integrated strategies toward effective, eco-friendly, and scalable PPCP mitigation.
Background Both chronic periodontitis and atherosclerotic cardiovascular disease are highly prevalent inflammatory diseases. The potential biomarker for these conditions is lactate dehydrogenase. The purpose of this study was to investigate salivary lactate dehydrogenase concentrations in patients with CP with and without ATH. Methods This was a cross-sectional study involving 87 participants aged 30-60 years, divided into four groups: the ATH-only group (n=25), the CP-only group (n=25), the ATH+CP group (n=25), and the normal control group (n=12). The participants underwent periodontal examination and sampling of unstimulated saliva. Salivary LDH was analyzed by ELISA. Statistical analysis included one-way ANOVA, post hoc tests, Pearson correlation, and ROC curve analysis. Results The salivary LDH values were significantly different among the groups ( p < 0.001). The ATH+CP group had the highest value of salivary LDH (24.55 ± 2.55 U/L), followed by the CP group (13.44 ± 4.27 U/L), the ATH group (11.68 ± 3.86 U/L), and the control group (1.53 ± 0.31 U/L). The values of salivary LDH were positively correlated with the clinical parameters of periodontal disease in the diseased groups. The diagnostic accuracy of the ROC analysis was excellent, with an AUC = 0.987. Discussion The results obtained in this study suggest that salivary LDH is potentially useful as a noninvasive biomarker for estimating inflammation in patients with periodontitis and cardiovascular disease. The increased levels of LDH, particularly among patients with combined conditions, suggest pathophysiological connections and support emerging evidence concerning the bidirectionality of associations between oral and systemic health. Conclusion Salivary LDH is a potential noninvasive biomarker of periodontal disease and may reflect the inflammatory burden in patients with concomitant ATH. The presence of elevated LDH in patients with both conditions indicates pathophysiological overlap.
Endophytes, microorganisms that live within plant tissues, are emerging as promising, eco-friendly solutions for both environmental remediation and sustainable agriculture. This review examines how these beneficial microbes can be leveraged to address the significant threats of soil and water pollution while reducing the agricultural sector's reliance on synthetic agrochemicals. Endophytes enhance phytoremediation by directly degrading or detoxifying pollutants and by improving the host plant's ability to tolerate stress and absorb contaminants such as heavy metals and pesticides. In agriculture, they boost plant growth by producing phytohormones and enabling biological nitrogen fixation, which in turn reduces the need for chemical fertilizers. They also strengthen plant resilience against biotic and abiotic stressors, such as drought, further diminishing the use of chemical inputs. Despite their considerable potential, challenges remain, including the lack of a centralized database for identification and a limited understanding of host specificity and the dynamics of endophyte communities under different environmental conditions. To fully realize the benefits of endophytes, future research must focus on comprehensive studies that explore their ecological interactions, model their behavior in plant tissues, and address the trade-offs between their positive effects and potential limitations. By integrating endophyte-based strategies, we can couple pollution mitigation with improved crop productivity, aligning with the principles of a circular bioeconomy and advancing sustainable development goals.
Marine-derived proteins and peptides are emerging as an exciting area of interest in terms of their potential for delivering improved performance through functional nutrition. Marine proteins, which have a good balance of essential amino acids, are easily digested and contain bioactive peptides that have antioxidant, anti-inflammatory, and metabolism-modulating properties. Recent scientific studies have found that marine protein can contribute to muscle building through the strengthening of connective tissues via collagen. Marine bioactive peptides are being studied to enhance vascular functions, reduce oxidative stress levels, and increase endurance and metabolism. This review outlines the existing literature on the biochemical properties, physiological pathways, and potential applications of marine proteins and peptides in sports nutrition, highlighting their increasing role in delivering enhanced performance through functional nutrition.
IntroductionResveratrol (3,4′,5-trihydroxy-stilbene), a naturally occurring polyphenolic stilbenoid present in grapes, red wine, peanuts, and berries, has garnered immense scientific interest over the last two decades due to its multifaceted pharmacological properties. Its active trans-isomer (RSV) has demonstrated antioxidant, anti-inflammatory, anti-diabetic, cardioprotective, neuroprotective, anti-aging, anti-obesity, and anticancer effects by modulating many key molecular targets. Despite encouraging preclinical findings, the clinical translation of RSV has been hampered by poor systemic absorption, rapid metabolism, and low oral bioavailability. MethodsThis systematic review adheres to the PRISMA 2020 guidelines and provides a comprehensive analysis of data obtained from PubMed, Scopus, Web of Science, Google Scholar, Google Patents, and ClinicalTrials.gov, spanning the period from 2000 to 2025. The focus was on patents and clinical trials investigating RSV and its associated chemicals. Studies were selected based on their relevance to RSV's pharmacokinetic changes, therapeutic efficacy, and disease-specific applications. ResultsWe evaluated 162 studies in their totality, including preclinical trials, individual clinical trials, and patent investigations. The findings suggest that RSV may be beneficial in the management of chronic problems, including diabetes, cardiovascular disease, neurological disorders, Non-Alcoholic Fatty Liver Disease (NAFLD), inflammatory skin conditions, and some malignancies. Nanocarriers, liposomes, structural derivatives, and dietary preparations are new ways to deliver drugs that have been proven to be more effective. Clinical investigations substantiate its benefits in glycemic control, vascular function, weight loss, and modulation of inflammation. Specific outcomes demonstrate variability among diverse populations and treatment protocols. DiscussionThe review stresses how RSV-based drugs are still getting better, especially with the addition of boronic acid analogues, combination formulations (such as those that include curcumin or flavonoids), and nanotechnology systems for administration. These groundbreaking ideas could improve stability, bioavailability, and function that is unique to certain diseases. Patent data show that RSV is more common in nutritional supplements, cosmetics, and drugs that are said to treat cancer, skin problems, digestive problems, and early aging. But there are still issues with dose optimization, differences among people, and regulatory constraints. ConclusionRSV and its compounds provide several therapeutic challenges while potentially facilitating holistic illness management. Their reliability, low cost, and diverse biological activities suggest significant promise for improving future healthcare. In addition, the growing number of patents and available clinical data further support their potential. However, continued research is needed to develop novel strategies for their field application and to better understand the relationship between their structure and biological activity.
IntroductionThe most essential factor in the micropropagation of Gapi banana plants is the induction of initial growth. The initial growth induction of cultures usually uses the growth regulators cytokinin and auxin or a combination of both, and can also utilize natural materials, such as Aloe vera and Vigna radiata, as a potential alternative to synthetic PGRs to induce shoots/callus/somatic embryos. This study aims to evaluate the effects of Aloe vera gel juice and Vigna radiata sprout extract, compared to the control media (MS-0), the BAP, and the combination of BAP with 2,4-D in Murashige and Skoog (MS) culture media in the micropropagation of Gapi banana. MethodsThe study was conducted as a laboratory experiment using Murashige and Skoog (MS) medium supplemented with Aloe vera gel juice (MS-LB) 10, 15, 20, and 25% (v/v), Vigna radiata sprout extract (MS-ET) 10, 15, 20, and 25% (v/v), and compared with control media MS-0, MS-BAP 1 mg L-1, and a combination of MS-BAP 1 mg L-1 with 2,4-D (0.1, 0.3, and 0.5 mg L-1). Data were analyzed using the Chi-square (χ2) test to determine differences among treatments. ResultsThe results were that the explants could grow to form the best shoots on 20% MS-LB medium and 10% MS-ET in the second month, and 1 mg L-1 MS-BAP medium formed several shoots in the third month. Adding a combination of 1 mg L-1 BAP + 2,4-D of 0.1 and 0.3 mg L-1 in MS media produced a compact callus. High concentrations of MS-LB medium (25%), 15 and 25% MS-ET media, and a combination of 1 mg L-1 MS-BAP + 0.5 mg L-1 2,4-D medium did not support explant growth because the inoculant remained blackened and did not develop. DiscussionThe findings of this study suggest that Aloe vera gel juice and Vigna radiata sprout extract can be considered as promising natural alternatives to synthetic plant growth regulators in banana micropropagation. This approach highlights the potential of eco-friendly and cost-effective culture media supplements to support sustainable plant tissue culture practices. ConclusionMS media added Aloe vera gel juice (MS-LB 20%, v/v) and Vigna radiata sprout extract (MS-ET 10%, v/v) were most effective in inducing shoot growth in the second month after inoculation, faster than 1 mg L-1 BAP treatment medium, which succeeded in forming shoots in the third month. The combination of BAP 1 mg L-1 + 2,4-D 0.1 and 0.3 mg L-1 created a compact callus from the first month. On the other hand, high concentrations of 25% (v/v) MS-LB, 15 and 25% (v/v) MS-ET medium, and the combination medium of 1 mg L-1 BAP with 0.5 mg L-1 2,4-D did not support explant growth because the explant remained blackened and did not develop.
IntroductionAging leads to reduced physical, mental, and social performance, lowering productivity. The fibroblast secretome, rich in bioactive proteins, growth factors, and enzymes, is essential for cell repair and regeneration. This study evaluated its regenerative potential by examining MEK1/2 pathway activation and type I collagen utilization to enhance senescent fibroblast proliferation. MethodsYoung (passage 3) and senescent (passage 24) fibroblasts were cultured in DMEM. Senescent cells were treated with 10% or 20% fibroblast secretome. Culture media were collected on days 1, 3, 5, and 7 for analysis of total protein, KGF, cell proliferation, MEK phosphorylation, and collagen concentration. Results The 20% secretome contained 5830,67 ± 181,62 µg/mL total protein and 1712,67 ± 7,19 pg/mL KGF. Senescent fibroblasts treated with 20% secretome showed a 44% proliferation increase on day 5 (21135,67 ± 1392,89 cells/well) versus controls (14637 ± 2250,57 cells/well, p<0,05). MEK1/2 phosphorylation rose from 0,01 ± 0,00 pg/mL at 45 minutes to 0,18 ± 0,03 pg/mL at 24 hours (p<0,05), indicating sustained activation. Discussion Secretome treatment enhanced MEK1/2 signaling and proliferation, with a 12% reduction in extracellular type I collagen on day 5 (2,67 ± 0,03 ng/mL vs. 3,02 ± 0,03 ng/mL, p<0,05), reflecting increased collagen utilization and matrix remodeling. ConclusionThe 20% fibroblast secretome enriched with KGF and bioactive proteins promotes senescent fibroblast proliferation via MEK1/2 activation and optimized collagen dynamics, supporting its potential in regenerative medicine for aging and tissue repair.
IntroductionDimethylsulfoniopropionate (DMSP) is a key marine organosulfur compound produced by phytoplankton and macroalgae that functions as an osmolyte, antioxidant, and precursor of dimethylsulfide (DMS)-a climate-relevant gas influencing the global radiation balance. Marine bacteria degrade DMSP through demethylation and cleavage pathways, driving the marine sulfur cycle. This study aimed to quantify DMS(P) concentrations and to isolate, identify, and characterize DMSP-degrading bacteria from the Cochin Estuary (CE), Kerala, India. MethodsSurface water and sediment samples were collected from fifteen CE stations across three seasonal regimes (pre-monsoon, monsoon, and post-monsoon) between 2015 and 2018. DMS(P) levels were determined by alkali hydrolysis followed by gas chromatography with headspace sampling. Heterotrophic bacterial abundance was estimated by spread plating on Zobell’s Marine Agar. DMSP-degrading bacteria were isolated on DMSP-enriched minimal medium, and selected isolates were identified by 16S rRNA gene sequencing. PCR amplification was performed to detect DMSP lyase (Ddd) genes, and phylogenetic analyses were conducted using MEGA6. ResultsDMSP concentrations ranged from BDL to 0.15 ng/µL in water and 0.01 to 2.35 ng/µL in sediments, with higher values recorded during the pre-monsoon season. A total of 112 water and 211 sediment bacterial isolates were obtained, with Gram-negative strains dominating (70% in water and 64% in sediment). Sediments harbored higher bacterial counts than water. Four isolates capable of growing on DMSP-enriched medium were identified: Acinetobacter calcoaceticus, Acinetobacter beijerinckii, Bacillus cereus, and Lysinibacillus fusiformis. Amplification of the dddP gene was observed in A. calcoaceticus. Seasonal variations in salinity, temperature, and nutrient levels influenced DMS(P) distribution, with higher concentrations recorded in sediments. DiscussionThe findings confirm CE sediments as microbial “hotspots,” dominated by γ-Proteobacteria and Firmicutes-groups known for their significant roles in sulfur cycling. Hydrographic seasonality, particularly salinity fluctuations, shaped bacterial diversity and DMSP transformation patterns. The detection of DddP genes suggests active enzymatic cleavage pathways contributing to atmospheric DMS release. ConclusionThis first baseline study on DMSP degradation in the CE highlights the ecological significance of estuarine sediments in sulfur cycling. The results enhance the understanding of microbial mediation of DMSP catabolism in tropical estuaries and its implications for climate regulation. Further research is warranted to elucidate additional catabolic pathways and environmental controls.
IntroductionIn an attempt to solve the problem of antifungal resistance in the Rhizopus species, which contributes to the severity of mucormycosis, the study was conducted to target the Heat Shock Protein 90 (HSP90). The main aim was to develop a new de novo protein inhibitor that was specific to the fungal HSP90 but not to the human counterpart, reducing off-target toxicity, and overcoming the limitations of the current therapies. MethodsA bioinformatics approach was adopted. The strategy involved determining the conserved domains of the HSP90 protein of different strains of Rhizopus through multiple sequence alignment. The ten possible de novo protein inhibitors were then generated using a deep learning model on the basis of the consensus sequence of this conserved region. The stability and binding affinity of these inhibitors were measured through molecular dynamics simulations and protein-protein molecular docking to the fungal and human HSP90 structures. ResultsThe analysis led to the identification of an inhibitor of lead de novo, Gen7, which exhibited improved binding and specificity. Molecular docking revealed that Gen7 had much higher affinity and interacted extensively with Rhizopus stolonifer HSP90 (12 hydrogen bonds, 4 salt bridges) than with human HSP90 (5 hydrogen bonds, 3 salt bridges). This fungal selectivity was later confirmed by subsequent molecular dynamics simulations. The Gen7-R. stolonifer complex was highly stable with an RMSD of about 4-5 A, whereas the Gen7-human complex was very unstable with a variation of RMSD of up to 15 A. DiscussionThe results indicate that deep learning and bioinformatics have the potential to be used in designing highly selective therapeutic agents. This will address the serious problem of off-target toxicity that has hampered the clinical development of earlier HSP90 inhibitors, offering a feasible solution to developing more effective and safer antifungal agents. ConclusionThis study successfully designed and computationally validated Gen7, a novel de novo inhibitor that selectively targets HSP90 in Rhizopus species. The research provides strong proof of concept for a new class of targeted antifungal agents, offering a promising avenue for developing innovative treatments against drug-resistant fungal infections like mucormycosis.
Introduction/ObjectiveThe aim of the study was to develop a technological approach for the production, storage, and preparation of strawberry microplants, facilitating their simultaneous acclimatization before planting in a greenhouse when favorable temperature and lighting conditions are established. MethodsThe initial material for the experiment was obtained via clonal micropropagation. Preparation, accumulation, and preservation of strawberry plants in vitro for delayed acclimatization were performed by periodically repeating the procedure of removing roots and leaves from previously rooted microplants and transferring them in the form of vegetating buds onto a fresh nutrient medium for further rehabilitation. The mass acclimatization of microplants was carried out under substrate-free flow-through hydroponic conditions, following which they were transplanted into a greenhouse in early spring. ResultsThe survival rate of microplants during the periodically repeated procedures of removing roots and leaves from previously obtained microplants reached 100% for all studied cultivars. Active leaf and root recovery began on days 3–7 and 10–14, respectively. A 1-month acclimatization period on flow-through hydroponics allowed for a 2–2.5-fold increase in microplant height, a 2-fold increase in root length, and an almost 3-fold increase in weight. DiscussionThe repetitive removal of microplant roots and leaves, with their further transplantation on fresh nutrient medium as vegetating buds for subsequent rehabilitation during periods of unfavorable environmental conditions, does not reduce the viability of microplants, allowing the preservation of planting material without loss until acclimatization is advisable. Acclimatization using flow-through hydroponics enables the elimination of fungal pathogenesis that occurs during the use of solid substrates. This protocol also increases the predictability of acclimatization outcomes and ensures successful transplantation into a greenhouse. ConclusionThe developed alternative strategy of strawberry clonal micropropagation allows prediction of the output volume of acclimatized microplants throughout a calendar year with a high degree of accuracy. The estimated quantity of such material can be compactly stored in a lighted tissue culture room, significantly saving time and resources. Microplants are well preserved until a favorable moment for acclimatization in a viable state as vegetating buds. After the rehabilitation cycles are complete, the microplants can be simultaneously transferred for acclimatization on flow-through hydroponics.
BackgroundThe emergence of multidrug-resistant (MDR) Stenotrophomonas maltophilia poses significant therapeutic challenges in clinical settings. This opportunistic gram-negative bacterium exhibits extensive antibiotic resistance mechanisms and forms biofilms on medical devices. Multiwalled carbon nanotubes (MWCNTs) have emerged as promising antimicrobial agents due to their unique physicochemical properties and multiple mechanisms of action. Materials and MethodsThirty clinical samples were collected from Kufa University Medical Center (November 2023-March 2024), yielding 20 confirmed S. maltophilia isolates through VITEK-2 identification. MWCNTs were synthesized via thermal decomposition and characterized using AFM, SEM, XRD, and FTIR. Antimicrobial activity was evaluated using well diffusion and broth microdilution methods at 25, 50, 75, and 100 μg/mL concentrations. Biofilm assays were performed using crystal violet staining. Results were compared with six conventional antibiotics. ResultsMWCNT characterization revealed 82.74 ± 5.2 nm grain size with armchair geometry. MWCNT suspensions demonstrated concentration-dependent antimicrobial activity, with 100 μg/mL producing 22.7 ± 1.58 mm inhibition zones, exceeding most antibiotics except ciprofloxacin (25.1± 4.98 mm). MIC values were 18.4 ± 10.01 μg/mL (MIC50 = 12.5 μg/mL). Biofilm formation was significantly reduced, with 100% MWCNT (The 100 μg/mL suspension) achieving 97.3% inhibition. All isolates exhibited high antibiotic resistance (75-95%). DiscussionThe superior antimicrobial activity of MWCNTs demonstrates their potential as alternative therapeutic agents for MDR S. maltophilia infections. Multiple mechanisms, including physical membrane disruption, reactive oxygen species generation, and metabolic interference, explain effectiveness against resistant isolates. The exceptional biofilm inhibition addresses critical clinical challenges, as S. maltophilia biofilms are difficult to eradicate with standard antibiotics. Clinically achievable MIC values support therapeutic feasibility. ConclusionMWCNT suspensions possess potent antimicrobial activity against MDR S. maltophilia with superior biofilm inhibition properties. The ability to overcome resistance mechanisms suggests MWCNTs represent promising alternatives for treating S. maltophilia infections, warranting further safety evaluation and clinical development.
Introduction Green nanotechnology utilises biological systems to produce nanoparticles, offering environmentally friendly and sustainable alternatives to traditional chemical methods. Methods Biosynthesis of zinc oxide nanoparticles (ZnO NPs) using novel probiotic bacteria from the gut of the snail Pila globosa-derived probiotics (Lactiplantibacillus plantarum)-has promising applications in the development of health-related feed supplements. ZnO:LP NPs represent a significant advancement in the biomimetic approach for enhancing biocompatibility, structural uniformity, and antimicrobial effectiveness. Results Characterization of the synthesised nanoparticles, conducted using techniques such as Ultraviolet-Visible-Near Infrared (UV-Vis-NIR) spectroscopy, X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM), confirmed the successful formation of a hexagonal wurtzite structure. Biological assays were performed to investigate the synthesized nanoparticles, revealing significant antibacterial and cytotoxic activities. Discussion This research highlights the promising applications of probiotic nanoparticles. Conclusion This research highlights the potential of probiotic-based nanoparticles for the development of comprehensive feed supplements with nutraceutical and nutri-biotechnological applications, as well as their implications for the food and healthcare industries.
Introduction/ObjectiveKyasanur Forest Disease (KFD), caused by the Kyasanur Forest Disease Virus (KFDV), is a tick-borne haemorrhagic fever endemic to South India and spreading to neighbouring states. The formalin-inactivated Chick Embryo Fibroblast (CEF) vaccine currently in use provides only short-term protection, requires repeated inoculations, and has limited coverage. A safe, simple-to-administer vaccine, which includes chills, fever, and headaches, was designed as a multi-epitope peptide vaccine (MEPV) against the immunodominant E protein of KFDV by using cutting-edge immunoinformatic and reverse vaccinology approaches. MethodsTen KFDV strains (1962–2016) were retrieved from NCBI and screened for antigenicity. The sequence of the E protein of the selected strain was screened for CTL, HTL, and B-cell epitopes using IEDB, NetMHCpan, and ABCPred. Predicted epitopes were evaluated for antigenicity, allergenicity, toxicity, immunogenicity, and conservancy across all the shortlisted strains. Potential epitopes were linked with suitable linkers to form the PKFDVac-I construct. Its physicochemical properties, structure stability, and immunogenic potential were evaluated using Expasy ProtParam, PSIPRED, AlphaFold, molecular docking with TLR-4, molecular dynamics simulation, and C-ImmSim immune simulation. ResultsSixteen epitopes (5 CTL, 3 HTL, 8 B-cell) cleared all screening criteria and were included in PKFDVac-I, a 279-amino-acid construct with a molecular weight of 29.16 kDa. The vaccine demonstrated high antigenicity, non-toxicity, non-allergenicity, solubility, and stability. Docking was found to be good, with a TLR-4 binding affinity of -1150.78 kcal/mol (Piper energy), supported by 387 non-bonding interactions. A 100-ns molecular dynamics simulation confirmed the stability of the complex. Immune simulation also anticipated robust humoral and cellular immunogenicity, higher antibody titers, long-lived persistence of memory cells, and robust IFN-γ induction. DiscussionPKFDVac-I had favorable immunological properties in silico. The design comprises conserved epitopes that are antigenic, safe, and immunogenic to the tested Indian KFDV strains from 1962 to 2016, ensuring lineage representativeness. Molecular docking and simulation reveal a stable interaction between receptors, and immune simulations predict durable adaptive immunity. ConclusionPKFDVac-I is a proposed multi-epitope peptide vaccine candidate for Kyasanur Forest Disease. The integration of diverse epitopes into a cohesive vaccine prototype demonstrates a promising avenue for custom synthesis and application in immunization strategies. The design represents a significant advancement in the evolution of KFD vaccines and warrants further in vitro and in vivo validation.
The production of biogas by the anaerobic decomposition of biomass guarantees an efficient and safe reduction of environmental pollution resulting from treating organic wastes, positioning it as a promising technology for the future. The anaerobic digestion process carried out in this research project utilized pumpkin peel, tamarind peel, and corn cob leaves as substrates. Cow, sheep, and poultry fecal wastes were also analyzed as microbial inocula for biogas production. This research focused on evaluating the physical, chemical, and bromatological parameters of the aforementioned organic waste to determine the optimal parameters that define the stages of biogas production. This study is proposed because Mexico is the main generator of this waste, and lacks studies on its proper management. The anaerobic digestion experiments were carried out in batch biodigesters with a capacity of 1.8 L at 36 ± 2 °C for a retention time of 30 days, using the methods of the Mexican Standards for Environmental Protection for the determination of humidity, ash, pH, Total Volatile Solids (TVS), Total Solids (TS), chemical oxygen demand, carbon, and nitrogen. Among the obtained results, the carbon/nitrogen ratio of the tamarind husk and corn cob leaves residues indicated a positive effect on biogas production in treatments mixtures 1 and 8, with values of 21.87 and 24.34, respectively, considered as values within the optimal range of 20:1 to 30:1, these values constitute the ideal availability of energy and nitrogen for cell replication of biogas producing methanogenic bacteria. These treatments were influenced by the Specific Methanogenic Activity (SMA) of the cattle manure inoculum, which had an organic matter biodegradability value of 0.58 grams of Chemical Oxygen Demand de Methane entre grams Total Volatile Solids for a day (COD-CH4/g TVS/day). This value supports the efficient assimilation of the carbon/nitrogen ratio based on the mentioned values. On the other hand, determining the TVS/TS ratio yielded percentages of 60%, 92%, and 95% in pumpkin peel, tamarind husk, and corn cob husks, respectively. Comparing the results with other studies, the optimal TVS/TS ratio should be greater than 50%; therefore, the results indicate that the substrates contain sufficient assimilable organic matter for anaerobic microorganisms and could produce biogas. Similarly, many scientific studies indicate that calculating the Specific Methanogenic Activity can not only determine the degree of biodegradability of substrates but also estimate possible pH changes in the medium, in addition to determining the maximum load of organic matter applied to any other type of anaerobic inoculum outside of this research work. Under these conditions, the treatments obtained a cumulative biogas yield of 3645 and 4250 milliliters of methane, respectively. This research contributes to improving the quality of biogas through the management of standardized waste.
Introduction The role of camel βeta-Nerve Growth Factor protein (Cam β-NGF), isolated from camel seminal plasma, in preserving sperm quality remains poorly defined. This study aimed to evaluate the impact of Cam β-NGF on mass motility, viability, and membrane integrity in dromedary (Camelus dromedarius) semen stored for 24 hours at 4 °C. Methods The semen samples were treated with varying concentrations of Cam β-NGF (10, 100, 500, and 1000 ng/mL) after preservation. Ten semen samples with mass motility ≥ 3 and viability ≥ 60% were selected, washed, suspended in HBSS, and treated with TCF-EY. After 24 hours of storage at 4 °C, samples were maintained at 36 °C with Cam β-NGF supplementation for 60 minutes. Mass motility, viability, and membrane integrity were assessed at T0, T30, and T60 minutes. Results The results revealed a significant decline in sperm quality traits after 24 hours of storage at 4 °C. However, mass motility was conserved after 30 minutes of incubation with 1000 ng/mL and 500 ng/mL of Cam β-NGF (p < 0.05). On the other hand, viability was significantly higher after 60 minutes of incubation with 1000, 500, and 100 ng/mL of Cam β-NGF compared to the control extender and 10 ng/mL supplementation (p < 0.05). Additionally, membrane integrity was significantly maintained at T30 and T60 minutes with 1000, 500, and 100 ng/mL of Cam β-NGF (p < 0.05). Discussion Short‐term storage at 4 °C markedly reduced dromedary camel sperm motility, viability, and membrane integrity, consistent with cold‐induced oxidative stress. Adding purified Cam β-NGF dose-dependently preserved these parameters over 1 h of incubation, indicating its value as a protective supplement for semen storage. Conclusion Cam β-NGF helps protect and maintain sperm traits during short-term storage; it seems to reduce alterations that might lower sperm quality. Thus, supplementing semen with Cam β-NGF may help maintain sperm quality, offering potential benefits for artificial insemination programs in dromedaries.
Introduction Current research emphasizes the endophytic fungi that produce secondary metabolites for biotechnological relevance. Botryosphaeriaceae species can produce significant bioactive compounds, and the Botryosphaeria genus is gaining attention in this regard. Aims This study aims to evaluate the bioactivity of Botryosphaeria species from Acacia species. Methods Botryosphaeriaceae species were isolated from Acacia karroo. Morphological and molecular identification was conducted using ITS and BOT primers. The dual method was utilised to determine the antifungal properties of the endophytic fungi. The PCR amplicons were sent for sequencing. Results The overall colonization rate reached 74%, with 18% attributed to the target species. Most isolates exhibited rapid growth, except for isolate 68. Nine isolates were identified as members of the Botryosphaeriaceae family via amplification with BOT primers. Isolate 13 showed strong antifungal effects against two test pathogens, while isolate 78 displayed moderate activity. BOT primer amplification yielded a 372 bp product. BLAST analysis identified isolate 13 as Botryosphaeria dothidea (MT197291.1; 92.76% identity) and isolate 78 as B. dothidea (AF027746.1; 98.41% identity). Endophytic fungi inhabit plant tissues asymptomatically, often engaging in mutualistic associations. Discussion This study confirms the high colonization of endophytic fungi and identifies Botryosphaeria dothidea with notable antifungal potential. Isolate 13 showed strong inhibition against test pathogens, suggesting possible biocontrol applications. Molecular and morphological tools effectively confirmed species identity. Limitations include the use of a single primer and limited pathogen screening. Conclusion The endophytic fungi isolated from Acacia karroo have the potential to be used as fungicides in the agricultural sector. These bioactive compounds have enormous prospects in the biotechnological sphere.
Introduction Glycerol, the main byproduct of biodiesel production, poses environmental challenges if not effectively utilized. Converting glycerol into bioethanol provides a sustainable route to support renewable energy development. This study explores the potential of microbial isolates with high lipase activity for efficient glycerol fermentation. Methods The tested strains included three bacteria (Serratia sp., Pseudomonas sp., Escherichia coli), one yeast (Saccharomyces sp.), and two fungi (Aspergillus sp., Trichoderma sp.). Morphological adaptations were evaluated using Scanning Electron Microscopy (SEM), and ethanol production was validated through Fourier-Transform Infrared (FT-IR) spectroscopy by identifying characteristic absorption peaks. Quantitative analysis of ethanol yield and glycerol conversion was conducted using High-Performance Liquid Chromatography (HPLC). Results SEM analysis confirmed structural adaptation of Serratia sp. and Saccharomyces sp. under fermentation stress. FT-IR analysis verified the presence of ethanol with an absorption peak at 3251.52 cm−1. HPLC results showed that Serratia sp. produced the highest ethanol yield of 17.83% (5.35 g/L) with a glycerol conversion of 40.33%, followed by Trichoderma sp. with a yield of 17.37% (5.21 g/L) and a conversion of 39.56%. Although E. coli exhibited the highest glycerol conversion (80.54%), its ethanol yield was low (1.88%), indicating diversion toward other metabolic pathways. Discussion These results highlight the superior adaptability and metabolic efficiency of Serratia sp. in channeling glycerol toward ethanol production. Structural stability under osmotic and ethanol stress supports its role as a robust bioethanol producer, while differences among species underscore the importance of strain-specific optimization. Conclusion Serratia sp. demonstrates strong potential for glycerol-to-bioethanol conversion, providing a promising candidate for sustainable biofuel production and biodiesel waste valorization.
