HAYATI Journal of Biosciences (HAYATI J Biosci; p-ISSN: 1978-3019; e-ISSN: 2086-4094) is an international peer-reviewed and open access journal that publishes significant and important research from all area of biosciences fields such as biodiversity, biosystematics, ecology, physiology, behavior, genetics and biotechnology. All life forms, ranging from microbes, fungi, plants, animals, and human, including virus, are covered by HAYATI J Biosci. HAYATI J Biosci published by Department of Biology, Bogor Agricultural University, Indonesia and the Indonesian Society for Biology. We accept submission from all over the world. Our Editorial Board members are prominent and active international researchers in biosciences fields who ensure efficient, fair, and constructive peer-review process. All accepted articles will be published on payment of an article-processing charge, and will be freely available to all readers with worldwide visibility and coverage. HAYATI J Biosci has been also indexed/registered in SCOPUS, Asean Citation Index, DOAJ, SINTA, National Library of Medicine Catalog, Crossref, ROAD, CABI, EBSCO, Publons, Scilit, Agricola, ProQuest and others.
Bogor Agricultural University, Indonesia
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Vital But Vulnerable: The Population Structure of Macaca maura in Bantimurung Bulusaraung National Park, South Sulawesi
Macaca maura is an endemic primate of South Sulawesi, currently listed as Endangered due to habitat fragmentation and increasing anthropogenic pressure. This study aims to analyze the population density, age structure, spatial distribution, and food types consumed by M. maura in the Karaenta Forest, TN Babul. This field study was conducted over four months (August–November 2024) using a grid-based home-range survey. The study area was divided into 1 × 1 km² grid plots; each encounter with M. maura was recorded with X–Y coordinates to visualize the movement patterns of each group. Data were collected through direct observation of eight groups (A–H) with a total of 193 individuals. Group sizes ranged from 13 individuals (Group F) to 39 individuals (Group A). Population density ranged from 52 to 134 individuals/km² with an average of 92 individuals/km². The age distribution, with an average IFR of 0.69, indicates that there is less than one infant per adult female in the M. maura population, suggesting a low reproductive rate. Most individuals were observed on the ground rather than in trees or on cliffs, indicating a preference for terrestrial habitats. Dietary records show considerable flexibility, with 27 plant species being utilized. These findings underscore the need for targeted habitat management strategies aligned with the species' ecological needs. This study provides baseline data to support future evidence-based conservation planning and environmental monitoring of M. maura.
Copyright (c) 2025 Warda Murti, Fatchur Rohman, Murni Saptasari, Ibrohim Ibrohim https://creativecommons.org/licenses/by-nc/4.0
Panicle Trait Diversity and Its Impact on the Productivity of Indonesian Foxtail Millet (Setaria italica L. Beauv.) Genotypes
Foxtail millet is a climate-resilient cereal crop that is growing in importance in food security and sustainable agriculture. This study aimed to identify panicle traits associated with yield, determine functional traits suitable for selecting high-productivity genotypes, and classify superior local Indonesian foxtail millet genotypes. Eight genotypes were evaluated using a randomized complete block design with three replications. Correlation analysis of 31 panicle traits revealed that grain filling rate, panicle weight, grain weight per panicle, grain weight per plant, and grain numbers on the middle and lower primary branches were positively associated with yield. Several of these traits also exhibited high broad-sense heritability and moderate GCV and PCV values, indicating their potential for selection. Principal component and two-way heatmap cluster analyses identified Buru, ICERI-7, Hambapraing, and Mauliru-2 as high-yielding genotypes. The findings provide critical insights for foxtail millet breeding programs in Indonesia focused on improving yield.
Copyright (c) 2025 Marisa Vidya Luthfiani, M Reza Pahlevi, Bambang Sapta Purwoko, Sintho Wahyuning Ardie https://creativecommons.org/licenses/by-nc/4.0
Arrowroot (Maranta arundinacea L.) Tuber Extract Modulated Gut Microbiota and Improved Glycemic Control in Type 2 Diabetic Mice
Type 2 diabetes mellitus (T2DM) is a metabolic disorder characterized by hyperglycemia and has become a global health concern. Gut microbiota alterations significantly influence diabetes onset, which can be mitigated through prebiotic intake. Arrowroot tubers (Maranta arundinacea L.) possess prebiotic potential due to their oligosaccharide and dietary fiber content. This study evaluated the effects of arrowroot tuber extract on blood glucose, glucose tolerance, and gut microbiota composition in a T2DM mouse model. Tubers were macerated in ethanol, and BALB/c mice were fed a high-fat diet for five weeks, followed by streptozotocin injection (35 mg/kg) to induce T2DM. Mice received 150 mg of arrowroot tuber extract orally for 28 days. Blood glucose levels were measured on days 0, 14, and 28, and an oral glucose tolerance test (OGTT) was performed. Cecal samples collected after four weeks underwent 16S rRNA sequencing, and data were analyzed using One-Way ANOVA (p<0.05). Treatment with arrowroot tuber extract for 28 days reduced blood glucose levels by 40.5% and improved glucose tolerance. The extract modulated gut microbiota, decreasing harmful bacteria while increasing short-chain fatty acid (SCFA)-producing genera such as Lachnospiraceae NK4A136, Ligilactobacillus, and Alistipes. These results indicate that arrowroot tuber extract can beneficially alter gut microbiota composition and ameliorate hyperglycemia, suggesting its potential as a prebiotic-based therapeutic strategy for managing T2DM.
Copyright (c) 2025 Artini Pangastuti, Putri Regita Anggraini, Shanti Listyawati, Siti Lusi Arum Sari https://creativecommons.org/licenses/by-nc/4.0
Preclinical Evaluation of HPV Type 52 L1L2 Chimeric Protein as a Cervical Cancer Vaccine Candidate
High-risk human papillomaviruses (HPVs) are the primary etiological agents of cervical cancer, accounting for more than 300,000 deaths annually worldwide. Current prophylactic vaccines based on recombinant L1 major capsid virus-like particles (VLPs) have demonstrated strong efficacy but are restricted to a limited spectrum of HPV types. To address this limitation, the present study evaluated a recombinant L1L2 chimeric protein derived from HPV type 52 as a potential candidate for a broad-spectrum vaccine. The chimeric protein was expressed in Escherichia coli strain BL21 (DE3) and purified for immunization studies. Female BALB/c mice (Mus musculus, n = 5 groups) were immunized, and immune responses were analyzed by enzyme-linked immunosorbent assay (ELISA) and pseudovirion-based neutralization assays (PBNA). The recombinant L1L2 vaccine candidate induced detectable antibody responses against HPV antigens; however, neutralizing activity remained modest. Histopathological analysis of liver and kidney tissues showed no evidence of toxicity, supporting the safety profile of the candidate. In summary, these results suggest that the HPV type 52 L1L2 chimeric protein represents a promising platform for the development of cervical cancer vaccines, although further optimization is required to achieve robust cross-neutralizing efficacy.
Copyright (c) 2025 Isti Kartika Sari, Joko Pamungkas, Apon Zaenal Mustopa, I Wayan Teguh Wibawan, Jendri Mamangkey, Sheila Chairunnisa, Herman Irawan, Ai Hertati, Nurlaili Ekawati, Rifqiyah Nur Umami, Ela Novianti, Maritsa Nurfatwa, Huda Shalahudin Darusman https://creativecommons.org/licenses/by-nc/4.0
A Comparative Study of Penicillin G Acylase Expression in Two Escherichia coli Strains: BL21 (DE3) and Arctic Express
The growing demand for semisynthetic beta-lactams has directed attention towards enzymes, specifically Penicillin G Acylases (PGAs), for their potential in synthesizing these antibiotics. This study delves into the expression of Achromobacter xylosoxidans PGA (AxPGA) in Escherichia coli, with a focus on enhancing the yield of active PGA, often constrained by a complex maturation process. The optimization of PGA expression included variations in IPTG concentration and the addition of CaCl2. Furthermore, the study compared PGA expression in E. coli BL21 (DE3) with that in E. coli Arctic Express (DE3), capable of co-expressing chaperones (chaperonin Cpn60 and Cpn10). Induction with 0.5 mM IPTG resulted in the highest hydrolytic activity in both strains, with Arctic Express (AE) exhibiting significantly higher activity due to improved folding facilitated by cold-adapted chaperonins. Alongside optimal IPTG induction, the addition of 10 mM CaCl2 in the culture media significantly increased PGA activity in both strains, highlighting that Ca2+ supplementation is an effective strategy to enhance the yield of functional PGA. Subcellular fractionation demonstrates that the periplasmic fraction yielded higher volumetric and specific activities compared to the cytoplasmic fractions in both E. coli strains, highlighting the importance of periplasmic processing for PGA maturation. This suggests that extracting the periplasmic fraction is an effective strategy for recovering active PGA while avoiding or reducing contamination either from co-expressed cytoplasmic chaperones or other intracellular proteins. These findings emphasize that induction strategy, ionic stabilization, and host strain selection play synergistic roles in increasing active recombinant PGA expression.
Copyright (c) 2026 Kartika Sari Cendana, Sri Rezeki Wulandari, Gabriela Christy Sabbathini, Maria Ulfah, Dini Achnafani, Abinawanto, Sunni Sofiah Aniqah, Is Helianti, Niknik Nurhayati https://creativecommons.org/licenses/by-nc/4.0
Characterization of Lignin Biosynthesis Encoding Gene, SiCOMT, from Nine Indonesian Foxtail Millet (Setaria italica (L.) P. Beauv.) Genotypes
Caffeic acid O-methyltransferase (COMT) is an essential enzyme that catalyzes the biosynthesis of lignin monomer units. Foxtail millet (Setaria italica) possesses three COMT-encoding genes. However, only SiCOMT1 and SiCOMT2 are considered expressed. This study investigates the characteristics of the two COMT-encoding genes across nine Indonesian foxtail millet genotypes. Phylogenetic analysis revealed that SiCOMT1 is closely related to N-methyltransferase genes, which are not involved in the lignin biosynthesis pathway. Meanwhile, SiCOMT2 is closely related to O-methyltransferase genes involved in lignin biosynthesis. SiCOMT2 from nine Indonesian foxtail millet genotypes exhibits 15 synonymous and three non-synonymous SNPs. SiCOMT2 amino acid showed Ala67Thr and Pro72Ala variations within the methyltransferase dimerization domain, and Glu146Asp within the O-methyltransferase domain. Among these, the Pro72Ala substitution is predicted to reduce the structural stability of the encoded protein. These findings suggest that SiCOMT2 may serve as a promising target for future genetic research and crop improvement strategies aimed at enhancing biomass quality by modifying lignin content and composition.
Copyright (c) 2026 Ramadaniarto Rizqullah, Nirwanti Saenal, M Reza Pahlevi, Willy Bayuardi Suwarno, Daisuke Tsugama, Sintho Wahyuning Ardie https://creativecommons.org/licenses/by-nc/4.0
Silicon Enhances Banana Resistance to Fusarium oxysporum f.sp. cubense through Ethylene Signaling Modulation and Root Tissue Protection
Fusarium oxysporum f.sp. cubense Tropical Race 4 (Foc TR4) is an agent of banana's vascular wilt that severely hampers production. It has been indicated that silicon (Si) might play a role in plant defense, but the banana–Foc pathosystem mechanism needs to be investigated. This study aimed to assess the impact of Si supplementation on banana plant responses to Foc infection, particularly using a completely randomized design arranged in four treatments: Control, Foc, Si, and Si*Foc. The transcript levels of the ethylene-receptor (EIN1, EIN3), ethylene-response factor (ERF1, ERF2), and the defense marker PR4 were quantified using qRT-PCR. Phenotypic observation, corm lesion severity, and root anatomy were also evaluated. Results indicated that early ethylene signaling (EIN1, EIN3) was down-regulated, and ERF2 was up-regulated in the presence of Si during Foc infection. Foc-infected plants treated with Si displayed smaller lesion areas, greater root structural stability, and less aerenchyma formation. Plant growth reduction caused by Foc was also relieved by Si, showing increased root-to-shoot biomass ratios and modified leaf shape. Multivariate analysis validated differences in physiological and transcriptional patterns in Si*Foc plants. These results indicate that Si improves banana resistance to Foc by regulating ethylene-mediated defense, tissue integrity, and stress resistance.
Copyright (c) 2025 Arief Pambudi, Yunus Effendi, Lisdar I Sudirman, Miftahudin https://creativecommons.org/licenses/by-nc/4.0
Exploring the Genetic Landscape of Octopus cyanea: Connectivity and Population Structure in Northern Waters of Sumatra
Octopus cyanea is an ecologically and economically important cephalopod with wide larval dispersal influenced by ocean currents. However, geographic isolation among island clusters in northern Sumatra may restrict gene flow and shape population structure. This study investigated the genetic diversity and connectivity of O. cyanea from five populations (Aceh Island, Weh Island, Banyak Island, Simeulue Island, and Nias Island) using mitochondrial cytochrome c oxidase subunit I (COI) sequences. A total of 21 individuals were successfully amplified, yielding 651 bp fragments. Sequence analysis revealed 11 haplotypes across populations, with haplotype diversity (Hd) ranging from 0.285 in Banyak Island to 0.800 in Aceh Island, and nucleotide diversity (π) between 0.001 and 0.009. Pairwise genetic distances varied from 0.000 to 0.004, with the highest divergence observed between Simeulue Island and Nias Island (0.004), while Aceh Island and Weh Island showed no differentiation. AMOVA indicated that most genetic variation occurred within populations (68.74%), with an overall Fst of 0.315, suggesting low to moderate genetic structuring. The haplotype network demonstrated shared haplotypes among four populations, whereas Simeulue Island harbored unique haplotypes, indicating partial genetic isolation. These results highlight substantial genetic connectivity among O. cyanea populations in northern Sumatra, facilitated by larval dispersal via ocean currents, but also emphasize localized divergence in Simeulue Island. The findings provide critical baseline data for developing ecosystem-based fisheries management, supporting shared stock strategies while accounting for population-specific conservation needs.
Copyright (c) 2026 Mutia Ramadhaniaty, Adrian Damora, Intania Moula, Nur Fadli, Nanda Muhammad Razi, Zainal A Muchlisin https://creativecommons.org/licenses/by-nc/4.0
Application of Lactic Acid Bacteria to Produce Bioactive Compounds from Tofu Waste Using Pineapple Bromelain Enzyme
Tofu is one of the main human foodstuffs and has become an alternative substitute for animal protein. Tofu contains relatively high protein, and active compounds, but the waste produced during the tofu processing process is very abundant, namely 3.5 million tons per year in Indonesia. The production of active compounds from tofu waste can use bromelain enzymes. The bromelain enzyme in this study was produced from the fermentation of pineapple leaf waste using Pediococcus pentosaceus E7. The production of active compounds from tofu waste has not been widely reported, so this study aimed to produce active compounds from tofu waste. This study began with bromelain production, measurement of bromelain activity, bromelain precipitation, peptide production from tofu waste, Liquid Chromatography-Mass Spectrometry (LC-MS) analysis, and antibacterial testing of active compounds derived from tofu waste degradation. The results showed that the peptide content of tofu liquid waste degraded using the bromelain enzyme was 41.01±0.06 mg/mL. The LCMS results showed that 18 bioactive compounds were contained in the degradation products of tofu liquid waste. The active compounds derived from the degradation of tofu waste are capable of inhibiting the growth of Salmonella Typhimurium ATCC 14028, yielding a clear zone index of 2.53±0.12.
Copyright (c) 2026 Zulfiqar Lukman, Dimas Andrianto, Anja Meryandini https://creativecommons.org/licenses/by-nc/4.0
Transcriptomic Dissection of Long-Juvenility in Soybean: A Comparative Study of Grobogan Cultivar and Its Derivative Long Juvenile AP18 Line
The long juvenile (LJ) trait in soybean extends the vegetative phase, enabling cultivation in low-latitude regions. This study investigated transcriptomic differences between the Grobogan cultivar (control) and the AP18 (LJ) soybean line using RNA-seq. Young leaf tissues from plants at the vegetative stage were collected under field conditions and sequenced using the Illumina NovaSeq 6000 platform. After quality filtering and alignment to the soybean reference genome, 24,716 expressed transcripts were retained for analysis. Differential expression analysis identified 5,005 upregulated and 4,534 downregulated transcripts in AP18 relative to Grobogan. Functional enrichment analysis revealed the upregulation of genes involved in ribosome biogenesis and secondary metabolism, and the downregulation of genes related to photosynthesis and energy metabolism. While known LJ-related genes FT2A (E9) and ELF3B (J) showed differential expression, the magnitude was modest. These findings highlight widespread transcriptomic reprogramming associated with the LJ trait, providing insight into potential pathways that influence juvenile phase extension.
Copyright (c) 2025 Habib Rijzaani, I Made Tasma https://creativecommons.org/licenses/by-nc/4.0
Bacterial Community in Apis cerana and Heterotrigona itama Honey Using a Metabarcoding Approach
Honey is a healthy, natural product with high nutritional value that is converted from sugar in nectar by bacteria in the honey stomach of the bees. Several beneficial bacteria in honey produce bioactive compounds, such as Lactobacillus in Apis mellifera honey, which synthesizes lactic acid, bacteriocins, and enzymes. Here, we employed the metabarcoding technique using the 16S rRNA gene to identify the bacterial community in honey from A. cerana and Heterotrigona itama collected in Sukabumi Regency, West Java, Indonesia. Genomic DNA from both honey samples was isolated using the ZymoBIOMICSTM DNA miniprep kit before sequencing with the Oxford Nanopore Technologies (ONT) platform. Our studies showed that the most dominant bacteria in the honey of A. cerana and H. itama were Paenibacillus glucanolyticus and Limosilactobacillus, respectively. In both types of honey, Gram-positive and Gram-negative bacteria were also detected, as well as lactic acid bacteria, including Acetilactobacillus jinshanensis and Limosilactobacillus. We also found Actinobacteria in A. cerana and H. itama honey. This genomic data showed that A. cerana honey has a higher bacterial diversity than H. itama. Our finding is the first genomic study of bacterial diversity found in the honey of A. cerana and H. itama that live sympatrically in a bee farm.
Copyright (c) 2025 Nurjanah Nurjanah, Rika Raffiudin, Yulin Lestari https://creativecommons.org/licenses/by-nc/4.0
Melissopalynological Analysis Revealed a Higher Pollen Diversity in Stingless Bees than in Honey Bees
Identifying floral resources used by honey bees and stingless bees is essential for sustainable beekeeping and understanding pollination ecology. This study compared the diversity of floral resources utilized by the honey bee Apis cerana and stingless bees Geniotrigona thoracica and Heterotrigona itama based on pollen in honey. Bee colonies were in the same Flora Nauli Beekeeping area, Pematang Siantar, North Sumatra, Indonesia, ensuring equal access to similar surrounding floral resources. Honey samples were collected, and pollen grains were extracted, acetolysed, and analyzed by counting 1,200 grains to determine frequency classes. Pollen diversity was calculated using the Shannon–Wiener index (H′). Stingless bee honey contained more diverse pollen than A. cerana, which only contained pollen from the Arecaceae family. Heterotrigona itama and G. thoracica honey contained pollen from 20 and 11 plant families, respectively. Each species had a distinct predominant pollen type (>45%): Cocos nucifera (A. cerana), Sapotaceae type (G. thoracica), and Casuarinaceae type (H. itama). A higher pollen diversity index was also observed in H. itama (1.05-1.83) than in G. thoracica (0.34–1.64) and A. cerana (0.66). These results indicate that stingless bees are more generalist than honey bees, highlighting their ecological role in supporting pollination networks in tropical agroecosystems.
Copyright (c) 2025 Tiara Sayusti, Rika Raffiudin, Nina Ratna Djuita, Dyah Perwitasari-Farajallah, Irmanida Batubara, Windra Priawandiputra, Rahmadi Sitompul, Svea Lina Jahnk, Hermann Behling https://creativecommons.org/licenses/by-nc/4.0
Plant-Derived Exosome-like Nanoparticles from Emprit Ginger (Zingiber officinale var. Amarum) and Its Potential Metabolite as Functional Food Ingredients
The Plant-Derived Exosome-like Nanoparticles (PDENs) are nano-sized vesicles secreted by plants that carry various bioactive compounds and have shown promise for use in functional food applications. This study investigated PDENs isolated from emprit ginger (Zingiber officinale var. Amarum), a local ginger variety renowned for its health-promoting properties. PDENs were extracted from rhizomes harvested at 8, 10, and 12 months, and analyzed for total phenolic content (TPC), total flavonoid content (TFC), and antioxidant capacity. The best-performing sample based on these parameters was selected for further metabolite profiling using LC-QTOF-MS. A total of 41 compounds were identified from the selected GDEN and ginger extract samples—32 compounds in the GDEN and 24 in the extract. These compounds belong to various groups, including amino acids, flavonoids, phenolics, lipids, terpenoids, vitamins, and others. Compound identification was based on public databases and literature concerning their potential as functional food ingredients. Among these, amino acids were the most abundant group in the GDENs, whereas phenolics were the dominant group in the ginger extract. This study underscores the potential of ginger-derived exosome nanoparticles as a rich source of bioactive compounds, supporting their further exploration and application in the development of functional food products. The balanced metabolite profile observed in GDENs highlights their unique advantages over conventional extracts. These findings reinforce the potential of emprit GDENs as promising candidates for functional food development.
Copyright (c) 2025 Paramita Sadha Dayinta Rukmi, Diah Ratnadewi, Christofora Hanny Wijaya https://creativecommons.org/licenses/by-nc/4.0
Biotransformation of the Textile Dye Reactive Black 5 with Aspergillus niger and Analysis of Ligninolytic Enzyme Activity
Removal of textile dye using biological processes is considered more cost-effective, generates no hazardous byproducts, and is ecologically friendly. This study analyzes the removal of the synthetic dye Reactive Black 5 (RB5) using the biological agent Aspergillus niger. The Treatment occurred in a 250 mL Erlenmeyer flask containing a mixture of dye solution. The preliminary research treatment involved varying dye concentrations, which were then analyzed by examining enzyme activity and fungal growth at different shaker rotations. The initial findings on concentration variation indicated that it was effectively removed up to approximately 90%. Further research determined that the highest concentration, 200 ppm, was used. The results obtained with rotation variations up to the sixth day were 95% for 100 rpm and 93% for 150 rpm. Dye removal utilizes laccase, lignin peroxidase (LiP), and manganese peroxidase (MnP) as degrading enzymes. The highest amount of enzyme activity was achieved at 100 rpm, corresponding to the most effective dye removal. The enzyme activity at the 100-rpm variation was observed for MnP (62.2 U/L), LiP (32.2 U/L), and laccase (16.6 U/L). The rapid growth of the Aspergillus niger fungus is associated with the biomass, which influences the biosorption mechanism. Biodegradation pathways indicate the cleavage of aromatic bonds, with toluene and propanoic acid identified as the final products.
Copyright (c) 2025 Diah Ayu Prawitasari, Edwan Kardena https://creativecommons.org/licenses/by-nc/4.0
Genome-Wide SNP Discovery and Population Structure Analysis of 244 Indonesian Local Rice Accessions
Single Nucleotide Polymorphism (SNP) markers are essential tools for the genetic management and breeding of rice, enabling precise characterization of genetic diversity. However, SNP datasets specifically developed for analyzing the population structure of Indonesian local rice remain limited. This study aimed to identify polymorphic SNP markers and analyze the lineage and population structure of 244 Indonesian local rice accessions. BAM files of these accessions were obtained from the 3K Rice Genome Project via the International Rice Informatics Consortium. High-quality SNPs were generated through variant calling and stringent filtering, followed by annotation and population analyses using IR64, DJ123, and Nipponbare as references. A total of 3,839,961 high-quality SNP loci were identified across the 12 rice chromosomes. Hierarchical clustering and principal coordinate analysis consistently grouped the accessions into three major clusters, corresponding to Indica, Japonica, and admixed groups. Population structure analysis confirmed two main subpopulations: Indica and Japonica, with Japonica further divided into Temperate and Tropical subgroups. Eleven accessions showed admixed ancestry. This study provides essential insights into the genetic diversity and population structure of Indonesian local rice, supporting genetic resource conservation, trait discovery, and breeding efforts.
Copyright (c) 2025 Rizky Dwi Satrio, Miftahul Huda Fendiyanto, Satya Nugroho, Aris Hairmansis, Miftahudin https://creativecommons.org/licenses/by-nc/4.0
Enhancement of α-Glucosidase Inhibitory Activity, Antioxidant Activity, and Antioxidant Compounds in Mulberry Leaves Under Salinity Stress
Salinity stress disrupts redox homeostasis in plants, leading to notable changes in the levels of bioactive compounds. These compounds are recognized for their significant role in inhibiting the α-glucosidase enzyme, which is relevant in glycemic control. Detailed insights into how salinity stress modulates α-glucosidase inhibition in mulberry leaves remain scarce. This study aims to investigate the effects of salinity stress on α-glucosidase inhibitory activity in mulberry leaf extracts, and to identify accessions exhibiting the most potent inhibitory properties. In vitro assays were employed to conduct qualitative phytochemical analyses and to quantify total phenolic content, total flavonoid content, and α-glucosidase enzyme activity. The results demonstrate that high salinity stress enhances the inhibitory activity of mulberry leaf extracts against the α-glucosidase enzyme. Notably, the MB2-3 sample exhibited the most favorable IC50 value (0.59 µg/mL), coupled with a substantial phenolic content (141.9 mg GAE/g extract) and the highest content of flavonoid observed among all tested samples (619.56 mg QE/g extract). Collectively, these findings highlight the capacity of salinity stress to augment both the inhibitory potential of mulberry extracts against α-glucosidase and the accumulation of phenolic and flavonoid compounds, with sample MB2-3 displaying auspicious attributes.
Copyright (c) 2025 Yasinta Ratna Esti Wulandari, Yohana Caecilia Sulistyaningsih, Agik Suprayogi, Min Rahminiwati, Triadiati Triadiati https://creativecommons.org/licenses/by-nc/4.0
Evaluation of Curcumin-derived Carbon-dots' Inhibitory Activity as SARS-CoV-2 Antiviral Candidate Using Chemical Crosslinking
In our previous work, we demonstrated that curcumin-derived carbon dots (Cur-CDs) have potential as antivirals for COVID-19. However, the precise mechanism of action remains unclear. This study investigated the potential of Cur-CDs against SARS-CoV-2 by targeting the dimerization of the C-terminal domain of nucleocapsid protein (N-CTD) using chemical crosslinking. Recombinant SARS-CoV-2 N-CTD was expressed, purified, and subjected to chemical crosslinking. The dimerization inhibition ability of Cur-CDs was assessed with ligand concentrations ranging from 0 to 2,000 μg/mL. Successful inhibition —defined as a noticeable reduction in SARS-CoV-2 N-CTD dimer band intensity on SDS-PAGE—was observed when Cur-CDs were present at 8 to 16 times the protein concentration. We hypothesize that Cur-CDs bind to the dimerization residues, preventing non-covalent interactions between monomers and limiting dimer formation. Our findings suggest that Cur-CDs could be a promising antiviral strategy for SARS-CoV-2, especially targeting the dimerization of the nucleocapsid protein. Additionally, this study also highlights the use of chemical crosslinking as a valuable tool for interaction-based drug screening.
Copyright (c) 2025 Audrey Angelina Putri Taharuddin, Nicholas Yamahoki, Rebecca Stephanie, Dian Fitria Agustiyanti, Popi Hadi Wisnuwardhani, Marissa Angelina, Yana Rubiyana, Ratih Asmana Ningrum, Andri Wardiana, Desriani Desriani, Hariyatun Hariyatun, Ferry Iskandar, Fitri Aulia Permatasari, Ernawati Arifin Giri-Rachman, Azzania Fibriani https://creativecommons.org/licenses/by-nc/4.0
Enhanced Lipase Production in Pichia pastoris via Multiple Copies of Bacterial Lipase Genes and Co-expression of the HAC1 Gene
The hac1 gene, a key regulator of the untranslated protein response (UPR), was co-expressed in Pichia pastoris GS115 to enhance the production of a lipase from Geobacillus stearothermophilus. Multicopy lipase constructs (1X and 4X) were transformed with pPICZAwbe_hac1, generating GS115/T1.2RQ(1X)_hac1 and GS115/T1.2RQ(4X)_hac1 strains. The GS115/T1.2RQ(1X)_hac1 strain showed an 186% lipase activity after 120 hours versus the control (100%), while the GS115/T1.2RQ(4X)_hac1 strain showed a faster initial increase (38% at 48 hours) and 28% at 120 hours, which was beneficial for efficient enzyme production. Overexpression of the hac1 gene enhances lipase production because it activates UPR genes when the endoplasmic reticulum is stressed due to a large number of recombinant proteins and forms proteins that are not appropriately folded. SDS-PAGE and tributyrin plate assays confirmed extracellular lipase expression (~43 kDa). These results demonstrate that hac1 co-expression significantly (p = 0.01) enhances lipase production in Pichia pastoris, especially in lower-copy constructs. This is the first report of co-expressing hac1 with Geobacillus stearothermophilus lipase genes in yeast. The findings are expected to contribute to developing more efficient microbial cell factories for producing industrial enzymes.
Copyright (c) 2025 Esti Puspitasari, Yepy Hardi Rustam, Andreas Adhi Satya, Antonius Suwanto, Aris Tri Wahyudi, Rika Indri Astuti https://creativecommons.org/licenses/by-nc/4.0
Protein Profile and Decolorization Potential of Copper-Resistant Klebsiella pneumonia CKJ 500 2.1.2 in Response to Textile Dyes and Copper
Heavy metals and synthetic dyes are major environmental pollutants, particularly in industrial effluents. The development of effective and safe bioremediation strategies to mitigate their ecological impact is therefore critical. In this study, the copper-resistant bacterium Klebsiella pneumoniae CKJ 500 2.1.2 was investigated for its capacity to decolorize textile dyes—specifically malachite green—and its associated enzymatic activities were characterized. Bacterial resistance was assessed using Luria–Bertani agar containing varying concentrations of copper and dyes. Decolorization efficiency was evaluated spectrophotometrically, protein expression was analyzed using SDS-PAGE, and dye degradation products were identified using gas chromatography–mass spectrometry (GC-MS). The strain exhibited high tolerance to both copper and dyes, achieving 99.4% decolorization of malachite green and 81% for Congo red. The presence of copper inhibited the decolorization of most dyes, except malachite green and methylene blue. SDS-PAGE analysis identified three key enzymes: laccase (~60 kDa), manganese peroxidase (~39 kDa), and azoreductase (~22 kDa). GC-MS revealed both toxic and non-toxic degradation intermediates, indicating partial detoxification. These findings highlight the potential of K. pneumoniae CKJ 500 2.1.2 for bioremediation of dye-contaminated effluents. However, further research is required to elucidate the complete enzymatic pathways involved and to ensure environmentally safe dye degradation.
Copyright (c) 2026 Wahyu Irawati, Reinhard Pinontoan, Jessica Marcellie, Andreas Valiant, Marcelia Sugata https://creativecommons.org/licenses/by-nc/4.0
Morphological and Genetic Variation of Filopaludina javanica (von dem Busch, 1844) (Gastropoda: Viviparidae) from Madura Island, Indonesia
Filopaludina javanica (von dem Busch, 1844) is a freshwater gastropod species in the Viviparidae family. Filopaludina javanica is widely distributed in freshwater waters in Java (including Madura), Sulawesi, Sumatra, Borneo, Papua, Thailand, and Vietnam. The morphological, morphometric, and molecular characterization studies of F. javanica based on the COI gene originating from Madura Island are still quite limited. Therefore, this study aims to determine the morphological variations, morphometry, and molecular characters of F. javanica in Bangkalan, Madura. The samples used are the Taxonomy Laboratory collection from Madura Island, followed by morphological observations, morphometry, and analysis of DNA: isolation, amplification, electrophoresis, and COI gene sequencing. Morphological and morphometric variations in F. javanica from Madura Island showed seven types of morphological variations. Principal Component Analysis (PCA) scatterplot results showed morphometric clustering of F. javanica based on morphological type. Identification of F. javanica using BLAST and comparison with the GenBank database revealed five nucleotide base variations, with an overall genetic distance of 0.031. Therefore, the phylogenetic tree shows that F. javanica from Madura Island belongs to the same clade as F. javanica from Sarawak, West Java, and North Kalimantan.
Copyright (c) 2025 Hafiidh Haqqu Nugroho, Reni Ambarwati, Nur Rohmatin Isnaningsih, Dwi Anggorowati Rahayu https://creativecommons.org/licenses/by-nc/4.0
Termites as Soil Engineers: A Study on Organic Carbon and Nutrient Dynamics using Baiting Techniques in Tropical Forest Ecosystem
This study explores how termite activity affects soil chemistry, those are Soil Organic Matter (SOC) and nutrient dynamics in pine and mahagony forest of varying ages in the Bromo Forest, Indonesia. Termite activity was assessed using wooden poles placed in PVC pipes as bait, which also served for soil sampling. The results showed significant differences in SOC and total NPK levels among different pole damage classes in each forest. The highest values were observed in pole damage class 4. Termite-influenced soil, especially those affected by the genera Macrotermes sp., Microtermes sp., and Schedorhinotermes sp., showed the highest concentrations of SOC and NPK (4.97%, 0.51%, 15.42 mg/100 g, and 45.9 mg/100 g, respectively). The termite diversity index showed moderate diversity in all pine forests and low diversity in mahogany forests. The termite diversity index indicated moderate diversity in pine forests and low diversity in mahogany forests, likely influenced by bait type. These results demonstrate that termite activity significantly enhances soil nutrient content and can be used as an indicator of soil fertility status in tropical forest ecosystems.
Copyright (c) 2025 Sri Rezeki Febriani, Dwi Priyo Ariyanto, Ongko Cahyono, Didi Tarmadi, Bramantyo Wikantyoso, Setiawan Khoirul Himmi, Muhamad Khoiru Zaki https://creativecommons.org/licenses/by-nc/4.0
Development of a Fluorescence Immunoassay Based on Curcumin Carbon Dots-labeled IgY Antibodies for SARS-CoV-2 Detection
Carbon dots (CDs) are widely utilized in biomedical applications as fluorescent labels for imaging and diagnostics due to their excellent biocompatibility and superior optical properties. These advantages often make CDs a substitute for organic fluorescent dyes, which suffer from low emission intensity and poor photostability when interacting with biomolecules. Moreover, carbon-based materials are eco-friendly and can be synthesized from natural sources, such as curcumin, a chromophore compound abundantly available in Indonesia. Therefore, this study conducted a preliminary investigation on curcumin CDs-labeled IgY antibodies (IgY-cur CDs) for fluorescence immunoassay of SARS-CoV-2 in rapid test applications. The synthesis of the fluorescent label involved a carbodiimide coupling reaction using EDC/NHS agents to conjugate IgY antibodies with curcumin CDs. The IgY-cur CDs conjugate was confirmed to detect antigens through FRET immunosensor mechanisms, showing a significant increase in fluorescence intensity with increasing antigen concentrations (p < 0.05), with a minimum sample concentration of 10 ng. Furthermore, the IgY-CDs cur conjugate was applied as a reporter in a fluorescence-based LFIA using a sandwich assay format. The test strip successfully detected synthetic multiepitope SARS-CoV-2 antigens with an estimated detection limit of 54.28 µg and nasopharyngeal samples from confirmed COVID-19 patients within 35 minutes of operation. The test strip was evaluated to be stable under cold storage at 4°C for up to 3 weeks. In conclusion, curcumin CDs-labeled IgY antibodies demonstrate promising potential for further development as fluorescent labels in rapid diagnostic applications targeting SARS-CoV-2.
Copyright (c) 2025 Nisa Amanda Rachmadani, Meilisa Keizia Soetomo, Audrey Angelina Putri Taharuddin, Arum Sinda Santika, Oktaviardi Bityasmawan Abdillah, Fitri Aulia Permatasari, Ferry Iskandar, Heni Rachmawati, Azzania Fibriani https://creativecommons.org/licenses/by-nc/4.0
Molecular Characterization of Anisakis spp.: Potential Source of Fish-borne Zoonosis in Coastal Living Environment in Semarang, Indonesia
Anisakis spp. are marine nematodes known to cause anisakiasis, a zoonotic disease transmitted through fish consumption with diverse clinical manifestations. In Indonesia, a country with extensive marine biodiversity and high fish consumption, data on anisakiasis remain limited. This study aimed to characterize the Anisakis spp. from the most significant commercial fish market in Semarang, Indonesia. A total of 17 Rastrelliger sp. were sampled. PCR amplification targeting the mitochondrial COX2 gene was used to identify the species of Anisakis spp. molecularly. Phylogenetic relationships, nucleotide diversity, and neutrality indices were assessed. The haplotype structures were visualized using the Haplotype Network. Subsequently, 7 of 17 (41.17%) Rastrelliger sp. samples were infected with A. typica. Molecular analysis revealed two species, A. typica and H. amoyense, with high haplotype diversity (1.00±0.016, diversity±SD). Phylogenetic analysis revealed two major clades: A. typica (Semarang and Southern Makassar) and H. amoyense (Semarang, China, and Bangladesh), with both species indicating high genetic connectivity. Neutrality indices suggested purifying selection and population expansion for both species (dN-dS = -5.017). These findings highlight the genetic variability and zoonotic potential of Anisakis spp. in the commercial fish market, emphasizing the need for surveillance of fish-borne parasitic infections in Indonesia.
Copyright (c) 2025 Derico Hitipeuw, Muhammad Muzhaffar Daud, Raisha Nuranindita, Sutra Diyana, Fariha Akmalina Amirudin, Aprillia Andika Rahayu, Edo Yungki Saputra, Nabila Gusvie Qanita, Surya Natanegara, Mutiara Rizqi Yuwanita, Ajeng Dwi Wusono, Martini Martini, Fauzi Muh https://creativecommons.org/licenses/by-nc/4.0
Enhanced Biodegradation of DDT by White-Rot Fungus Phlebia brevispora TMIC34596 in a Nutrient-Rich Medium at Short Incubation Time
1,1,1-Trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) is a persistent organic pollutant that remains a global environmental concern due to its high toxicity and recalcitrance. This study investigated the ability of the white-rot fungus Phlebia brevispora to degrade DDT in a nutrient-rich Potato Dextrose Broth (PDB) medium under a short incubation time. The fungus degraded 64.25% of DDT within 7 days. Metabolites identified by GC/MS were DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), DDD (1,1-dichloro-2,2-bis(4-chlorophenyl) ethane), and DDMU (1-chloro-2,2-bis(4-chlorophenyl) ethylene), indicating both reductive dechlorination and dehydrogenation pathways. The results demonstrate that P. brevispora has strong potential for rapid biotransformation of DDT in nutrient-rich environments.
Copyright (c) 2025 Erly Grizca Boelan, Adi Setyo Purnomo, Fajriah, Ichiro Kamei, Maria Aloisia Uron Leba, Hildegardis Missa, Anselmus Boy Baunsele, Aloysius Masan Kopon https://creativecommons.org/licenses/by-nc/4.0
Assessing Climate Factors and Cyanobacterial Abundance on Microcystins Prediction Using Artificial Neural Network: A Case Study in Malaysia’s Drinking Water Reservoir
Toxic cyanobacterial blooms often lead to contamination with cyanotoxins, particularly microcystins. This study aims to examine microcystins persistence in a selected public water supply system and predict their concentration at various points based on climate factors and cyanobacterial abundance. Using the Enzyme-Linked Immunosorbent Assay (ELISA) method, microcystins concentrations were quantified at various points of the water supply system, including the raw water intake, reservoir, water treatment plant outlet, and distribution system. The highest microcystins concentration was detected at the reservoir with a mean concentration of 2.63 μg/L. An artificial neural network (ANN) model was developed to predict microcystins concentration. Rainfall, temperature, chlorophyll-a, phycocyanin (BGA-PC), and mcyE gene copy numbers were used as inputs, while microcystins concentrations at various water sampling points served as outputs of the multilayer perceptron ANN. Using the Statistical Package for the Social Sciences (SPSS, ver. 29), three networks with scaled conjugate gradient, sigmoid functions, and one hidden layer with 4 to 13 neurons were trained and validated to determine the best configuration that fits the observed data. The result shows a satisfactory prediction at the reservoir (Point 2) with low values of error (root mean square error = 0.065) and high coefficient values (R2 = 0.894) between experimental and predicted values, which are below the maximum value of the actual concentrations. Phycocyanin (BGA-PC) and chlorophyll-a had the most positive effects in predicting microcystins concentrations. These results indicate that ANN modelling can be a reliable tool for predicting microcystins contamination in drinking water reservoir.
Copyright (c) 2025 Nurul Awatif Ahmad, Som Cit Sinang, Nurul Hila Zainuddin, Noorazrin Abdul Rajak https://creativecommons.org/licenses/by-nc/4.0
Osteological Development of Yellow Rasbora Fish Rasbora lateristriata (Bleeker, 1854) Treated with Mangosteen Garcinia mangostana L. Peel Simplicia
Mangosteen peel simplicia (Garcinia mangostana L.) contains potential ingredients used in medicine. A toxicity test of mangosteen peel simplicia is needed before it is used. Wader pari (Rasbora lateristriata) is a potential animal model for toxicity testing. Thus, this study aims to determine the effect of mangosteen peel extract at various concentrations on the behavior and bone structure of R. lateristriata. In this research, fish behavior was observed at 48 hpf (hours post-fertilization) and 30 dpf (days post-fertilization). Bone structure observed at 96 hpf, 7 dpf, 14 dpf, 21 dpf, 28 dpf, 35 dpf, and 42 dpf using the Alizarin Red ‒ Alcian Blue method. Observation data on behavior and bone structure were analyzed descriptively, while the number of vertebrae was analyzed quantitatively using One-Way ANOVA (P < 0.05). The results showed that the ossification of cranium and caudal complex skeleton components in the 1 and 5 μg/mL treatments was significantly behind that of the control and 0.5 μg/mL treatments. Furthermore, 5 μg/mL treatment caused some behavioral and swimming pattern abnormalities in 48 hpf larvae. In conclusion, exposure to 1 and 5 μg/mL mangosteen peel simplicia resulted in skeletal growth inhibition in R. lateristriata. Exposure to 5 μg/mL mangosteen peel simplicia caused abnormalities in the behavior and swimming pattern of 48 hpf larvae.
Copyright (c) 2025 Pradnya Paramita, Luthfia Uswatun Khasanah, Bambang Retnoaji https://creativecommons.org/licenses/by-nc/4.0
Modification of the Kato-Katz Diagnosis Technique Using “Canang” Flower Waste Extract as a Staining for Ascaris lumbricoides Eggs
Helminthiasis, particularly that caused by Ascaris lumbricoides, is a major global health issue, especially in areas with poor sanitation. One method for preventing and controlling Ascaris lumbricoides infection is through identification via the Kato-Katz diagnostic technique. The use of synthetic dyes, such as methylene blue, in the Kato-Katz method raises concerns for both human health and the environment. In humans, methylene blue causes skin irritation, gastrointestinal issues upon ingestion, and systemic effects. Furthermore, its environmental impact includes reducing light penetration and acting as a toxic component in food chains. An alternative approach involves utilizing post-use offerings from Hindu rituals in Bali, known as canang, which consist of flower components such as Impatiens balsamina L. and Tagetes erecta. The natural dyes found in these flowers serve as an alternative to traditional staining methods. This study examined the efficacy of flower extracts as stainings using the cellophane absorption test, helminth egg detection and morphology identification, pH test, and measurement of heavy metal concentration. The dye made from canang flower waste at 3% did not differ much from manufactured stainings. Thus, canang flower waste is a safe alternative.
Copyright (c) 2025 Kadek Indira Maheswari, Putu Sathiya Adi Janendra, I Komang Tri Yasa Widnyana, Dewa Gede Putra Mahayana, Komang Kirana Ardhia Pramesti, Indra Dwisaputra, Made Bayu Permasutha, Nyoman Intan Permatahati Wiguna, Made Kurnia Widiastuti Giri https://creativecommons.org/licenses/by-nc/4.0
Spatial Variability of Water and Sediment Quality in Pond Outlet: Implications for Coastal Ecosystems in Mangrove Areas, Pasuruan, East Java
Aquaculture expansion in Pasuruan's coastal areas raises concerns about water and sediment pollution, but the spatial variability and magnitude of these impacts remain insufficiently quantified. This study aimed to assess the water and sediment quality at pond outlets across different aquaculture systems. Samples were collected from seven intensive (high-input shrimp farming), traditional (low-input, extensive ponds), and silvofishery (integrated aquaculture with mangroves) aquaculture farms from July to September 2024, coinciding with peak farming activity. Water and sediment samples were collected from seven farms and analyzed for chemical parameters, organic matter content, and redox potential. Results of the current research record extensive spatial and temporal heterogeneity that is driven by aquaculture management and external events like the WSSV epizootic in August 2024. Ammonia concentrations increased dramatically at stations within intensive vannamei shrimp farms (5.5 mg/L), while downstream stations exhibited natural dilution and mitigation by mangroves. Sediment quality analysis demonstrated a reducing condition with redox potential values from as low as -100 mV and SOM concentrations as high as 23%. Correlation analysis highlighted intensive farming systems as the main drivers of water and sediment degradation, with mangrove belts demonstrating resistance through nutrient adsorption and filtration with COD, TSS, and organic matter as primary pollution contributors. These findings emphasize the urgent need for adopting sustainable practices, such as polyculture systems, reduced feed input strategies, the implementation of constructed wetlands, and enhanced mangrove rehabilitation around aquaculture zones, to minimize environmental impacts and preserve coastal ecosystem health.
Copyright (c) 2025 Putri Nurhanida Rizky, Atika Marisa Halim, Andina Chairun Nisa, Ega Aditya Prama, Ulfauza, Muhammad Khusni Oktahimawan
Impact of Nicotine and Nicotine-Free E-Cigarettes on Body Weight, Erythrocyte Dynamics, Sperm Morphology, and Molecular Outcomes in Mice
The effect of electric cigarettes (e-cigs) on health should be a concern since their legality has occurred in Indonesia. Young adults use e-cigarettes not as therapy but as a lifestyle. The vapor component contains propylene glycol, vegetable glycerin, flavor, and nicotine, which may harm human health. This study examined the health risks of e-cigarettes on body weight, erythrocytes, and sperm morphology in mice. We used twenty healthy male mice exposed to the cigarette's vapor for thirty days. Mice were divided into room air as the negative control, nicotine-free, and 18 mg nicotine-containing groups. Our results showed that the body weight of the nicotine-free and nicotine-containing groups increased, while micronuclei formation increased after e-cig exposure. Furthermore, abnormal sperm morphology in the nicotine-containing groups increased significantly, yet the nicotine-free group was not significantly different compared to the negative control. These results were supported by the molecular prediction that exposure to e-cigs might increase oxidative stress and decrease the expression of TGF-β. Moreover, studies on it are ongoing to gather scientific evidence.
Copyright (c) 2025 Andita Ayu Mandasari, Siti Ayu Maryam, Chilyatul Firdausi, Muhammad Iqbal Maulana, Muhammad Hardi Kurnia, Dyah Ayu Widyastuti, Dheasy Herawati
Entomological Assessment of Mosquito Diversity and Density in Negara District, Bali: Implications for Vector-Borne Disease Surveillance
Indonesia has one of the highest prevalences of malaria in Asia, with annual case numbers on the rise. Despite being a major international tourist destination, the Negara Sub-district in Jembrana Regency, Bali, continues to experience notable incidences of both diseases. Entomological surveys are crucial for evaluating mosquito diversity and density, which supports effective vector control strategies. This study, conducted in five villages from September to October 2024, employed entomological survey methods to calculate the House Index (HI), Container Index (CI), Breteau Index (BI), and Density Figure (DF). Mosquito species diversity was assessed using the Shannon-Wiener diversity index and evenness (E) and dominance (C) metrics. Results indicated that HI, CI, and BI values were within the low transmission risk category, with a DF of 4 and a larva-free rate (ABJ) of ≤95%. Six mosquito species were identified: Aedes aegypti, Aedes albopictus, Anopheles dirus, Armigeres subalbatus, Culex quinquefasciatus, and Mansonia uniformis. The species diversity was moderate (1 < H' ≤ 3), even species distribution (E > 0.6), and low dominance (C ≤ 0.5). These findings suggest a relatively low risk of vector-borne disease transmission. However, given the presence of multiple disease-vector species and the region's dynamic human activity, continuous entomological surveillance remains essential to anticipate population surges and strengthen disease prevention efforts.
Copyright (c) 2025 Dani Sari Widyawati, Maharani Retna Duhita https://creativecommons.org/licenses/by-nc/4.0