Muhammad Zakaria¹, Sayed Hussain^2*, Essa Ali³, Jawad Munawar Shah⁴, Syed Hassan Raza⁵, Maqsood Wazir⁶, Abid Ali², Muhammad Shehzad¹

¹Department of Agronomy, Faculty of Agriculture, The University of Poonch, Rawalakot, Azad Jammu and Kashmir, Pakistan

²Department of Horticulture, Abdul Wali Khan University Mardan, KPK, Pakistan

³Department of Food Science and Technology, Zhejiang University of Science and Technology, China

⁴College of Agriculture, Bahaudin Zakarya University, Pakistan

⁵Institute of Crop Sciences, Zhejiang University, Hangzhou, China

⁶Department of botany, Karakoram International University, GB, Pakistan

*Corresponding author’s email: sayedhussain@awkum.edu.pk

Received: 28 August 2025 / Revised: 05 November 2025 / Accepted: 11 November 2025 / Published Online: 09 December 2025

Abstract

Drought stress is one of the most relevant abiotic stress factors globally, greatly reducing the crop productivity of agricultural crops. Maize globally is an important economic cereal crop, and it is sensitive to water deficit, which is an important problem of the semi-arid areas of Pakistan. Since to test them under drought stress, the current research aimed to investigate different morphological, physiological, and biochemical traits linked to drought tolerance in maize genotypes and conducted in the laboratory of the Department of Agronomy, Faculty of Agriculture, The University of Poonch, Rawalakot, Azad Jammu and Kashmir, Pakistan. Currently two maize varieties TP-1217 and Fakhr-NARC were exposed to different PEG (5% and 10%) to study their performance. The results show that root fresh weight was higher (1.76g) in Fakhr-NARC at PEG 5% that was followed by TP-1217 with 1.72g. The shoot fresh weight of 1.63g and 1.76g observed under PEG 5%. The photosynthesis rate of 17.66% in TP-1217 and 20.32% in Fakhr-NARC under PEG 5% was observed which was significantly reduced to 11.76% and 18.33% at PEG 10%. The chlorophyll contents of 7.15 and 13.69 were observed in PEG 5% which was further reduced to 4.79 and 9.1 at PEG 10%. The relative water contents (26.74% and 43.02%) were higher in TP-1217 at both PEG concentrations. However, the sugar (38.57), proline (52.22mgg^-1, 694.75mgg^-1), and antioxidant enzymatic activities were significantly higher in Fakhr-NARC at PEG 10%. Whereas membrane integrity (18%), H₂O₂ (18.2) and MDA (37.99) contents were higher in TP1217.

Keywords: Antioxidant enzymes, Chlorophyll, Drought stress, Maize, Polyethylene glycol

Phan-Phuong-Trang Huynh¹, Thanh-Tri Do², Thanh-Cong Nguyen³, My-Ngoc Bui³, Tuan-Loc Le⁴, Thanh-Luu Pham⁵, Hoang-Dung Tran^1*

¹Faculty of Biology and Environment, Ho Chi Minh City University of Industry and Trade (HUIT), 140 Le Trong Tan Street, Tay Thanh ward, Ho Chi Minh City 72009, Vietnam

²Faculty of Biology, Ho Chi Minh City University of Education, 280 An Duong Vuong street, Cho Quan ward, Ho Chi Minh City 72820, Vietnam

³Institute of Applied Research and Technology Transfer HUFI, Ho Chi Minh City University of Industry and Trade, 93 Tan Ky Tan Quy Street, Tan Son Nhi ward, Ho Chi Minh City 72011, Vietnam

⁴Department of Biotechnology, Faculty of Applied Science and Technology, Nguyen Tat Thanh University, 1165 National Road 1A, Ho Chi Minh City, Vietnam

⁵Faculty of Environment and Labour Safety, Ton Duc Thang University, 19 Nguyen Huu Tho Street, Tan Hung ward, Ho Chi Minh City 700000, Vietnam

*Corresponding author’s email: dungth@huit.edu.vn

Received: 27 August 2025 / Revised: 13 November 2025 / Accepted: 21 November 2025 / Published Online: 29 November 2025

Abstract

Phycocyanin is a blue pigment–protein with antioxidant properties, but its use in foods is limited by poor stability. A cyanobacterium, Leptolyngbya sp. CCAH036/1, was isolated from the Can Gio Mangrove Biosphere Reserve, Vietnam, and identified by morphology and 16S rRNA sequencing (96.5% similarity to the closest reference). C-phycocyanin was extracted by freeze–thaw and lysozyme treatment and then encapsulated with maltodextrin using spray drying. The optimized powder contained 20.53 mg/g phycocyanin, 5.14% moisture, 79.43% encapsulation efficiency, and 56.66% antioxidant retention. After three weeks of storage at 4 °C, both pigment content and antioxidant activity remained above 80%. Heating at 50–70 °C preserved about half of the activity, but stronger heat caused rapid decline. Stability was also greatest at pH 5–7. The powder was added to sticky rice mooncakes at 5–20%. At 15% supplementation, the cakes contained 1.874 mg/g phycocyanin and 43.78% antioxidant activity, with no loss of texture or sensory quality. The results indicate that spray drying with maltodextrin is recommended as an effective approach to stabilize phycocyanin from the local Leptolyngbya strain for incorporation into functional foods processed at moderate temperatures and near-neutral pH.

Keywords: Phycocyanin, Leptolyngbya sp., Microencapsulation, Spray-drying, Antioxidant, Mooncake

Sahiruddin Sahiruddin^1*, Muhammad Yusuf¹, Athhar Manabi Diansyah¹, Masturi Masturi¹, Herdis Herdis², Tulus Maulana³, Syaputra Wibowo⁴

¹Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea Makassar, South Sulawesi 90245, Indonesia

²Research Center for Animal Husbandry, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta, Bogor 16915, West Java, Indonesia

³Research Center for Applied Zoology, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta, Bogor 16915, West Java, Indonesia

⁴Eijkman Research Center for Molecular Biology, National Research and Innovation Agency (BRIN), Cibinong, Bogor 16911, Indonesia

*Corresponding author’s email: sahirsabile@unhas.ac.id

Received: 12 September 2025 / Revised: 04 November 2025 / Accepted: 11 November 2025 / Published Online: 21 November 2025

Abstract

Semen quality is one of important factor that impacts cattle reproduction. However, the seminal plasma and spermatozoa separation is still unclear. We attempted proteomic and metabolomic techniques on fresh bovine semen and its compartments to molecularly characterise and cross these two compartments. The proteomic profile of the semen plasma showed pronounced accessory proteins enhancing lipid binding, ion homeostasis, and membrane dynamics, particularly PDC-109, enolases, VDAC2, and SP-10, while the spermatozoa comprised scaffolding, anchoring, and mitochondrial enzymatic proteins such as AKAP3, tektins, cylicins, and COX5B. The dichotomy was reinforced by complementary metabolomic analysis, with seminal plasma containing antioxidants and lipids such as taurine, ergothioneine, palmitoylglycine, and stearamide, while spermatozoa were enriched in metabolites associated with energy, including citrate, inosine, succinic semialdehyde, and pantothenic acid. Pathway analysis reinforced plasma specialisation about antioxidants and lipids rather than spermatozoa on glycolysis and amino acid metabolism with oxidative phosphorylation. Collectively, these results illustrate the presence of interconnected but non-interchangeable molecular domains in which seminal plasma provides protective and regulatory buffering, while spermatozoa are specialised in structural and energetic components for fertilisation. The candidate biomarkers identified from this study such as PDC-109, AKAP3, cylicins, taurine, and citrate illustrate molecular outputs corresponding to the quality of semen and provide a systematised context for enriched understanding of bovine reproductive biology.

Keywords: Bovine semen, Metabolomics, Seminal plasma, Spermatozoa, Proteomics

Prathan Luecha¹, Attachai Trunjaruen^2,3, Theeraphat Kongnok², Wipa Yaowachai⁴, Pattaraphorn Panomai⁵, Pitakpong Maneerattanarungroj⁶, Nisarat Tungpairojwong², Monthira Monthatong², Worasitikulya Taratima^2*

¹Department of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Science, Khon Kaen University,

Khon Kaen 40002, Thailand

²Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand

³Department of Biology, Faculty of Science, Burapha University, Chon Buri 20131, Thailand

⁴Queen Sirikit Botanic Garden, the Botanical Garden Organisation, Chiang Mai 50180, Thailand

⁵Health Science and Aesthetic Program, Faculty of Science and Technology, Rajamangala University of Technology Krungthep, Bangkok 10120, Thailand

⁶Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand

*Corresponding author’s email: worasitikulya@gmail.com

Received: 25 August 2025 / Revised: 15 October 2025 / Accepted: 01 November 2025 / Published Online: 21 November 2025

Abstract

Phytolacca americana L. (pokeweed) Root contain valuable pharmaceutical compounds but their slow growth and invasive spread make large-scale cultivation difficult. The purpose of this study was to optimize conditions for the enhancement of secondary metabolites while examining the effects of NaCl, MeJA, and SA on the amount of phenolics, flavonoids, and saponins in pokeweed callus cultures. Significant interactions between elicitor concentration and duration were observed for all phytochemical parameters (p < 0.05). MeJA at 100 µM for 3 days and SA at 150 µM for 9 days produced the highest phenolic contents (14.26 and 15.82 µg/mg DW, respectively; p < 0.001) and enhanced antioxidant activities (DPPH, ABTS, FRAP). NaCl at 200 mM for 21 days maximized EsA content (16.73 µg/mg DW), surpassing natural roots and all other treatments (p < 0.001). Phenolic content correlated positively with antioxidant capacity, indicating phenolic compounds as major contributors to antioxidant activity. In contrast, EsA showed negative correlations with other parameters, suggesting its accumulation was independent of phenolic biosynthesis and antioxidant properties. All elicitors upregulated biosynthetic genes via signaling and transcription factor activation. Overall, NaCl, MeJA, and SA effectively enhanced bioactive compounds in pokeweed callus cultures, offering a promising alternative for metabolite production without relying on natural plant resources.

Keywords: Abiotic elicitor, Antioxidant activity, Elicitation, Esculentoside A, Saponin

Hung Nguyen Thanh¹*, Tra Mai Huong², Thu Tran Thi Anh³, Thanh Pham⁴

¹Faculty of Biotechnology, Ho Chi Minh City Open University, Ho Chi Minh City, Vietnam

²Lac Hong University, Dong Nai City, Vietnam

³Thu Dau Mot University, Ho Chi Minh City, Vietnam

⁴Faculty of Biology, University of Education, Hue University, Hue City, Vietnam

*Corresponding author’s email: hung.ngt@ou.edu.vn

Received: 19 September 2025 / Revised: 05 November 2025 / Accepted: 12 November 2025 / Published Online: 20 November 2025

Abstract

This study aimed to evaluate the effects of different selenium (Se) fertilization methods on yield, quality, and organic Se accumulation in pak choi (Brassica rapa L.). A two-year small field experiment (2024–2025) was conducted with four treatments: control (CK), soil application (T1), foliar application (T2), and combined soil plus foliar application (T3). Results indicated that Se treatments significantly increased yield by 4.9–17.55%. Se concentration increased by 175.71–305.71% in root and by 202.81–314.08% in stem. Selenomethionine (SeMet) was the dominant organic form while selenocysteine (SeCys) also increased markedly. Crude protein, ash, and micronutrients (Fe, Zn, and Ca) were enhanced, especially under T3. Overall, the combined soil and foliar application (T3) proved most effective for improving yield, nutritional quality, and organic Se accumulation, offering a sustainable strategy for Se biofortification in leafy vegetables.

Keywords: Biofortification, Brassica rapa L., Nutritional quality, Selenium fertilization, Selenium speciation, Soil and foliar application

Shahad Alsunusi¹, Taha A. Kumosani², Etimad Huwait², Khalid O. Abulnaja², Soonham S. Yaghmoor², Said S. Moselhy³*

¹Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia

²Experimental Biochemistry Unit, King Fahd Medical Research Center and Production of Bio-products for Industrial Applications Research Group, King Abdulaziz University, Jeddah, Saudi Arabia

³Biochemistry Department, Faculty of Science, Ain Shams University, Cairo, Egypt

*Corresponding author’s email: moselhy6@hotmail.com

Received: 16 August 2025 / Revised: 24 October 2025 / Accepted: 31 October 2025 / Published Online: 20 November 2025

Abstract

Herbicides are used worldwide for protection the crops from weeds, but they still pose health problems. We investigated the mechanism of sweet basil (Ocimum basilicum L) methanolic extract (OBLE) against neurotoxicity induced by malathion in rats through the modulation of neurotransmitters, redox changes, cytochrome c release, and apoptosis. This study included sixty male Sprague Dawley rats allocated into six equal groups: Group I, Control; Group II, rats given  malathion  dissolved  in DMSO at the LD50/10 dose (10 µg/kg b.w) orally for 4 weeks; Group III, rats given DMSO orally (0.1 ml/kg b.w) for 4 weeks; Group IV, normal rats treated with OBLE (100 mg/kg) for 4 weeks; Group V (Preventive group), rats administered  malathion   and OBLE orally (100 mg/kg) for 4 weeks; Group VI (treated group), rats administered  malathion   for 4 weeks followed by OBLE for another 4 weeks. The phenolic and flavonoid contents of OBLE were found to be 85 mg GAE /g and 82.65 mg catechin E/g, respectively. Rats given malathion showed a significant reduction in brain acetylcholine esterase (AchE) activity, levels of serotonin, noradrenaline, dopamine and elevation of GABA compared to normal rats. However, rats in the preventive or treated groups with OBLE exert a significant elevation in serotonin, dopamine, and NE levels and activation of AChE activity while GABA level decreased compared with untreated.  Malathion   induced brain tissue Cyt c release, reduced apoptosis markers (caspase 3 and 9) and increased mitochondrial membrane potential (marker of mitochondrial function). OBLE was found to reduce mitochondrial potential, enhance antioxidant activity, reduce the release of Cyt c, and enhanced caspase 3 and 9. The molecular docking analysis showed the potential interactions between the tested compounds target proteins and receptors is reflected by the binding free energies (G) (measured in kcal/mol), with a lower value indicating a more stable interaction. It was concluded that, the phenolic and flavonoid content of OBLE contributed to the neuroprotection against malathion induced neurotoxicity via neurotransmitters, redox status and apoptosis.

Keywords: Malathion, Neurotoxicity, Apoptosis, Redox status, Ocimum basilica  

Fatima Aslam Bhatti^1*, Muhammad Naveed¹, Hafiz Naeem Asghar¹, Shahzad MA Basra², Wajid Ishaque³

¹Institute of Soil & Environmental Sciences, University of Agriculture Faisalabad-38040, Faisalabad, Pakistan

²Department of Crop Physiology, University of Agriculture Faisalabad-38040, Pakistan

³Nuclear Institute for Agriculture and Biology (NIAB), P.O. Box 128, Jhang Road Faisalabad, Pakistan

*Corresponding author’s email: fatimaaslambhatti@gmail.com

Received: 07 September 2025 / Revised: 03 November 2025 / Accepted: 11 November 2025 / Published Online: 19 November 2025

Abstract

Endophytic diazotrophic bacteria from non-host crops represent a promising source for developing novel bioinoculants. This study isolated and characterized nitrogen-fixing bacteria from the leaves and twigs of Populus euphratica and assessed their ability to colonize and promote growth in Wheat. From fifty-two isolates obtained on nitrogen-free media, five potent strains—identified by 16S rRNA sequencing as Beijerinckia fluminensis FA-7, Stenotrophomonas maltophilia FA-9, Pseudomonas aeruginosa FA-16, Klebsiella pneumoniae FA-4, and Enterobacter cloacae FA-8 were selected for their nitrogen fixation and plant growth-promoting (PGP) traits. These strains exhibited multiple PGP activities, including phosphate solubilization, indole-3-acetic acid (IAA) production, and the production of siderophores and exopolysaccharides. In growth chamber and wirehouse experiments under varying nitrogen regimes [full (N+), limiting (N-), and zero (N0)], all strains enhanced at least one wheat growth parameter. Notably, Beijerinckia fluminensis FA-7 significantly improved early growth, nutrient uptake, and root architecture—increasing root length, surface area, and fine root development—as confirmed by WinRhizo analysis. Our findings demonstrate that endophytic diazotrophs from Populus can successfully colonize wheat and enhance its growth, highlighting their potential as effective microbial inoculants for agriculture.

Keywords: Endophytes, PGPR, Driazotroph, Nitrogen fixation, Wheat

Muhammad Yusuf^1*, Sigit Bintara², Widjiati Widjiati ³, Athhar Manabi Diansyah¹, Sahiruddin Sahiruddin¹, Masturi Masturi ¹, Kurniawan Dwi Prihantoko², Aeni Nurlatifah², Suryo Kuncorojakti³, Suhernik Susilowati³, Tulus Maulana⁴, Syahruddin Said⁴

¹Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea Makassar, South Sulawesi, Indonesia

²Faculty of Animal Science, Gadjah Mada University, Depok, Sleman Regency, Special Region of Yogyakarta, Indonesia

³Faculty of Veterinary Medicine, Airlangga University, Campus C, Mulyorejo St., Surabaya, East Java, Indonesia

⁴Research Center for Applied Zoology, National Research and Innovation Agency, Jl. Raya Bogor Km. 46, Cibinong, West Java, Indonesia

*Corresponding author’s email: myusuf@unhas.ac.id

Received: 06 August 2025 / Revised: 11 October 2025 / Accepted: 14 October 2025 / Published Online: 18 November 2025

Abstract

Reproductive efficiency in livestock can be enhanced through sperm sexing technologies; however, conventional methods such as flow cytometry are expensive, technically demanding, and impractical for field use. Simpler, affordable, and biologically validated alternatives are crucial, especially for indigenous breeds like Bali cattle. This study evaluated the effectiveness of a freeze-dried albumin (egg white) gradient (10%–30%) method for separating X- and Y-bearing spermatozoa in Bali bulls, alongside proteomic profiling to confirm the biological distinctiveness of each fraction. Semen from three mature Bali bulls (Bos javanicus) (n = 3; 5 ejaculates per bull; total 15 ejaculates) underwent separation via the freeze-dried albumin gradient. Sperm quality and kinematic parameters were analyzed using Computer-Assisted Sperm Analysis (CASA), while morphometric assessment estimated sexed sperm proportions. Proteomic analysis using liquid chromatography–tandem mass spectrometry (LC-MS/MS) was followed by gene ontology enrichment, hierarchical clustering, and Pearson correlation, with identified proteins validated against literature. The method enriched X-spermatozoa in the upper layer (69.67%) and Y-spermatozoa in the lower layer (73.50%), with significant differences p < 0.05 in motility, viability, and membrane integrity. Proteomics identified 418 proteins, including 45 unique to X-sperm and 159 unique to Y-sperm. GO enrichment linked X-sperm proteins to nuclear and structural roles, while Y-sperm proteins were associated with mitochondrial and motility processes. Clustering distinctly separated the two sperm types, and several proteins correlated strongly with functional traits. Literature validation confirmed sex-specific markers.  This method represents a low-cost, biologically validated alternative for practical sperm sexing in Bali cattle, combining sperm quality assessment with proteomics to support sex-preferential breeding

Keywords: Bali bull, Bos javanicus, Sperm sexing, Albumin gradient, Proteomics, Sex-specific proteins