Malaysian Applied Biology

The Complete Mitochondrial Genome of the Parasitic Leech Pterobdella arugamensis (Annelida: Piscicolidae) from Malaysia and Phylogenetic Insights within Subclass Hirudinea

2026-09-29T17:07:16+00:00

The Therapeutic Potentials of Malaysian Medicinal Plants for Cardiovascular Diseases: A Review

2026-09-20T05:24:49+00:00

Cardiovascular diseases are among the most common diseases around the world with a considerably high rate of mortality. In view of this, multiple approaches have been proposed, especially in exploiting the potential of herbal plants due to their availability, accessibility and medicinal values. This review was conducted based on 16 medicinal herbs available in Malaysia as listed in the Malaysian Herbal Monograph. The aim was to study their effects on the alleviation of cardiovascular diseases, to analyse the relationship between the phytochemical compounds of herbal plants in relation to cardiovascular diseases and to review their underlying mechanism of action. The medicinal plants are organised based on their effects on alleviating cardiovascular diseases either directly or indirectly through reduction of cardiovascular disease risk factors, such as antihyperlipidemic, vasodilatory effects, cardioprotective and anticoagulant effects. Both in vivo and in vitro studies are included in this review to demonstrate the scientific value of related plants in cardiovascular diseases. In conclusion, many studies have identified a diversity of herbal plants in Malaysia to possess cardiovascular-relieving activities. Further studies can be conducted to develop these plants for cardiovascular disease treatment or prevention.

Anti-HHV-1 Mechanism and Pharmacokinetic Profile of Bioactive Compounds from Phaleria macrocarpa (Scheff.) Boerl Fruit

2026-05-18T23:20:10+00:00

This study reports on Phaleria macrocarpa fruit protein aqueous extract (PMFPAE) compounds, the mechanistic anti-human herpes virus type-1 (HHV-1) insights, and pharmacokinetic profiles. The mechanistic insight of PMFPAE treatment on the virus UL27 gene expression that encodes the viral attachment glycoprotein, gB, was determined to be downregulated at all replication phases, especially at 8 hr post-treatment, concurring with the late replication phase. Seven main water-soluble compounds in PMFPAE were identified via liquid chromatography-tandem mass spectrometry (LC-MS/MS). These include organic acids (D-gluconic acid), xanthones (mangiferin, homomangiferin), and flavonoids (quercetin, myricetin 5-methyl ether, oroxylin A, and apigenin 7-glucuronide-4′-(6′‘-malonylglucoside)). All the compounds have drug-likeness properties according to Lipinski’s Rule of Five except for apigenin 7-glucuronide-4′-(6′‘-malonylglucoside). Except for quercetin and oroxylin A, the compounds have low gastrointestinal absorption, indicating low possibility for oral use. ProTox-II predicted D-Gluconic acid as the least toxic, and quercetin has the highest toxicity. CarcinoPredEL indicated none of the compounds to be carcinogenic. In molecular docking studies, two compounds, homomangiferin and apigenin 7-glucuronide-4’-(6′‘ malonylglucoside), showed strong binding affinity towards virus glycoprotein gB. Five compounds have strong binding towards the virus glycoprotein gD, including mangiferin, myricetin 5-methyl ether, homomangiferin, apigenin 7-glucuronide-4’-(6′‘ malonylglucoside), and quercetin. Homomangiferin can bind to both virus attachment glycoproteins gB and gD, with additional antioxidant and anti-inflammatory activities. Strong binding affinities of mangiferin and homomangiferin to site I (subdomain IIA) of human serum albumin (HSA) suggest effective compound transport within the bloodstream, enhancing their stability and half-life. In conclusion, PMFPAE consists of multifacet drug-like bioactive compounds that inhibit viral entry by binding to gB and gD and have antioxidant and anti-inflammatory activities. The predicted pharmacokinetic profile suggests that compounds of PMFPAE are more suitable for development as an anti-HHV topical agent due to the limited gastrointestinal absorption, toxicity of several key compounds, and limited binding to HSA.

The Effects of Plant Growth Regulators, Carbohydrate Substrates and Photoperiod on Shoot, Root and Callus Induction from Rhizome Shoot Bud Explants of 'Bentong' Ginger (Zingiber officinale Roscoe)

2026-09-20T04:34:30+00:00

Zingiber officinale Roscoe from Bentong, also known as the ‘Bentong’ ginger, is a premium Malaysian-grown ginger recognised for its distinctive aroma, strong pungency, and elevated levels of 6-gingerol and 6-shogaol. In vitro platforms such as callus and cell suspension cultures represent a promising approach for the production of novel secondary metabolites from this valuable crop. This study aims to assess the effects of plant growth regulators, carbohydrate substrates and photoperiod in the induction of friable callus from rhizome bud explants of Z. officinale Roscoe from Bentong for subsequent studies on the establishment of cell suspension cultures. Explants were subjected to surface sterilisation and cultured in treatments of 2,4-D and kinetin, different carbohydrate substrates and photoperiods. Results in this study indicated that treatment with 2.5 mg/L 2,4-D resulted in 93.33% callus induction and 0.257 ± 0.06 g fresh weight, whereas treatment with 2.0 mg/L kinetin resulted in 85.71% callus formation, with 0.106 ± 0.02 g callus fresh weight. Both treatments of 2,4-D and kinetin produced yellowish white friable callus. The concentration of 30 g/L sucrose was considered the optimal carbohydrate level for callus induction in rhizome bud explants of ‘Bentong’ ginger. Additionally, explants cultured under continuous dark conditions achieved the highest percentage of callus induction (70.0%) with the highest callus fresh weight (0.270 ± 0.07 g). This study successfully induced friable callus from rhizome bud explants of ‘Bentong’ ginger (Z. officinale Roscoe), providing a suitable foundation for subsequent callus and cell suspension culture studies aimed at producing valuable secondary metabolites from this ginger cultivar.

Comparative Analysis of Protease-Assisted Extraction and Three-Phase Partitioning for Protein Recovery from Thai Black Soybean Okara

2026-09-20T04:34:28+00:00

Okara, a by-product of soy-based food production, has gained increasing interest, particularly that derived from black soybean, due to its higher protein and nutrient contents than yellow soybean. However, its utilization remains challenging due to its high perishability. Limited studies have addressed okara bioactive composition, bioactivity, and the environmental impacts of different extraction methods. This study aimed to compare environmentally friendly extraction methods and evaluate their efficiency and functional properties for recovering bioactive proteins from Sukhothai 3 black soybean okara, using protease-assisted extraction (Protease-P6SD & Protease-A2SD) and three-phase partitioning (TPP). Extraction yield, amino acid composition, and antioxidant activity were evaluated. TPP showed a significantly higher extraction yield (26.61 ± 2.24%) than protease-assisted extraction (16.49 ± 0.95% and 15.36 ± 1.63% for Protease-P6SD and Protease-A2SD, respectively) (p<0.05). In contrast, the amino acid composition analysis showed that extracts obtained by protease-assisted extraction contained a broader range of essential amino acids than those obtained by the TPP method. This may be attributed to the higher specificity of proteolytic enzymes toward target proteins compared with chemical extraction methods. Antioxidant assays (DPPH and ABTS) revealed significantly lower IC₅₀values, approximately two-fold lower, in protease-derived extracts compared to TPP (p<0.05), indicating stronger antioxidant activity. These findings demonstrate that a higher extraction yield does not necessarily correspond to superior protein quality. While TPP provides a higher extraction yield, protease-assisted extraction enhances the amino acid profile and bioactivity. Therefore, enzymatic extraction represents a promising green approach for enhancing the utilization of black soybean okara. This study highlights the potential of Sukhothai 3 black soybean okara as a valuable biofunctional ingredient for functional food applications.

Acute Toxicity Effects of Heavy Metals to Juvenile Climbing Perch, Anabas testudineus (Bloch, 1792) Within 96-Hour Exposure

2026-09-20T04:34:27+00:00

This study examines the acute toxicity effects of four heavy metals, arsenic (As), lead (Pb), cadmium (Cd), and zinc (Zn), on juvenile climbing perch, Anabas testudineus, under controlled laboratory conditions. Analytical-grade arsenic acid, cadmium sulfate, Lead(II) chloride, and zinc sulfate were used in preparing standard stock solutions. Fish mortality was recorded at 24-hr intervals over 96 hr. Median lethal concentrations (LC₅₀) and times (LT₅₀) for the juveniles were predicted using cumulative mortality, based on probit analysis run in Microsoft Excel. The water physical parameters, including pH (6.57), temperature (27.1–28.2°C), and dissolved oxygen (6.3–7.2 mg/L), were maintained within optimal ranges, ensuring that fish mortality and behavioural changes were mainly affected by metal toxicity. Arsenic recorded the highest toxicity (LC₅₀= 0.17 ppm), with 100% mortality at 1.0 ppm within 48 hr. Lead (Pb) demonstrated a narrow tolerance range, exhibiting no mortality at concentrations below 1.35 ppm but resulting in rapid 100% mortality at 3.41 ppm (LC₅₀= 2.47 ppm). Cadmium is toxic with sudden, lethal effects at higher concentrations (LC₅₀=2.5 ppm). Zinc became toxic at higher levels (100% mortality at 89 ppm, LC₅₀= 50.35 ppm). The fish toxicity was in the following order: As > Pb > Cd > Zn. An inverse relationship has been detected between LC₅₀values and metal concentration, highlighting the concentration- and time-dependent nature of toxicity. Unusual fish behaviour, including erratic swimming and excessive mucus production, indicated neurophysiological stress responses. This ranking (As > Pb > Cd > Zn) highlights arsenic as the most dangerous contaminant to A. testudineus in aquatic systems.

Bromelain-Enriched Diet Improves Morphology and Spawning Performance in Clarias macrocephalus

2026-09-20T05:24:45+00:00

This study aimed to investigate the effects of a bromelain-supplemented diet on the external morphological and reproductive characteristics of Clarias macrocephalus, and to assess how these changes influence spawning performance. Functional food enzymes are attracting increasing interest in aquaculture due to their potential to improve digestion and physiological performance. Bromelain, a proteolytic enzyme derived from pineapple, was evaluated for its effects on external morphology and reproductive characteristics. The experiment comprised two dietary treatments: a control diet and a bromelain-supplemented diet. External morphology was assessed by examining skin pigmentation, tail curvature, mucus secretion, muscle tone and reproductive traits such as genital papillae development and abdominal distension. External morphologies were scored using a standardised scale of 1–5, and statistical analyses were performed using independent t-tests and chi-square tests. Fish fed the bromelain diet showed significantly greater skin pigmentation (3.65 ± 0.49 vs. 1.15 ± 0.37), increased tail curvature (3.40 ± 0.60 vs. 1.60 ± 0.50), higher mucus secretion (3.80 ± 0.89 vs. 1.50 ± 0.51) and reduced muscle tone (3.40 ± 0.60 vs. 1.25 ± 0.55) compared to the control group (p<0.001). In contrast, reproductive characteristics including genital papilla structure and abdominal distension showed no significant differences (p>0.05), although successful spawning and a high fertilisation rate (87.4 ± 5.2%) were observed in the bromelain-fed pairs. The findings suggest that bromelain supplementation has a marked effect on the external morphology and physiological condition of catfish, which could enhance spawning readiness despite limited visible changes in reproductive anatomy. This indicates that the effects of bromelain have an internal or metabolic potential. The study highlights the potential of bromelain as a feed additive in aquaculture and recommends further research to investigate its effects on reproductive hormones, gonadal histology and synergistic interactions with bioactive compounds to optimise reproductive outcomes in C. macrocephalus culture.

Comparative Impact of Drying Techniques on Physicochemical, Proximate Compositions, and Antioxidant Properties of Roselle Calyces (Hibiscus sabdariffa L.) for Sustainable Agriculture

2026-09-20T05:24:47+00:00

This study evaluated the impact of different drying methods, freeze drying (-120°C), food dehydrator (65°C), and oven drying (65°C) on the quality of roselle calyces to extend their shelf life and availability, considering implications for sustainable agriculture. Key quality parameters assessed included physicochemical properties, proximate composition, and antioxidant activity. Freeze drying produced the highest quality results, closely resembling fresh roselle in several aspects. The colour analysis showed freeze-dried samples had the highest chroma values with L* (35.97), a* (31.86), and b* (17.3), indicating vibrant colour retention. The pH of freeze-dried roselle was the lowest at 2.74, indicating a desirable balance of acidity. In terms of proximate analysis, freeze-dried roselle had the lowest moisture content (7.55%) among dried samples, which is crucial for extended shelf life. It also had the highest ash content (8.06%), protein content (11.06%), and crude fat content (2.51%), highlighting its superior nutrient retention compared to other drying methods. Oven-dried roselle showed higher ash (8.69%) and fat (1.44%) contents but had a lower moisture content (6.36%) than dehydrator-dried roselle, which had the highest moisture content (8.30%) and the lowest ash (5.75%) and protein (5.86%) contents. Antioxidant analysis revealed that freeze drying preserved the highest levels of antioxidant activity (14.22% RSA), total phenolic content (12.3 mg GAE/mL), total flavonoid content (44.9 mg QE/mL), and ascorbic acid content (22.267 mg/100 g). These findings highlight the effectiveness of freeze-drying in maintaining the bioactive compounds and overall nutritional quality, which is crucial for sustainability agriculture practices aimed at preserving crop value and extending product availability.

Effects of Dehydration and Roselle Powder Addition on Physicochemical Characteristics, Antioxidant Properties and Hydroxymethylfurfural Formation in Stingless Bee Honey

2026-09-23T12:03:35+00:00

High moisture in stingless bee honey increases the risk of fermentation, which may adversely affect the quality. This study evaluates the effects of dehydration and roselle powder addition on the physicochemical characteristics, hydroxymethylfurfural (HMF) formation, antioxidant properties, and sensory acceptance of stingless bee honey. The study was conducted using a 4 × 2 factorial design. The factors investigated were dehydration treatment (food dehydrator at 55°C for 18 h) and roselle powder addition at four levels (0%, 5%, 10%, & 20%). Physicochemical analyses included moisture content, colour intensity, pH, total soluble solids, hygroscopicity, and HMF content. Total phenolic and flavonoid contents were determined, and antioxidant activity was evaluated using the Ferric Reducing Antioxidant Power (FRAP) assay. Sensory evaluation was conducted with 30 panelists to assess multiple sensory attributes. All data were analysed using two-way ANOVA. Both factors had significant effects on the physicochemical characteristics (p<0.05). HMF content increased significantly following dehydration and roselle powder addition (p<0.05). Antioxidant activity, phenolic and flavonoid contents increased significantly with both dehydration and roselle powder addition (p<0.05). Sensory evaluation revealed that overall acceptance remained at a moderate liking level, with no significant differences (p>0.05) observed among the samples. In conclusion, controlled dehydration combined with roselle powder addition improved the physicochemical properties and antioxidant characteristics of stingless bee honey while maintaining comparable sensory acceptance to the control samples. However, dehydration also increased HMF formation, indicating that further optimisation of the dehydration process is required.

A Laboratory Framework for Evaluating Antimicrobial Activity in Herbal Soaps: A Model System Using Neem and Turmeric Formulations

2026-09-23T12:03:33+00:00

The rising concern of skin infections caused by common microbes like Staphylococcus aureus and Candida albicans exacerbated by increased antimicrobial resistance from overuse of conventional drugs has increased the public’s interest in natural and herbal-based products. While commercial medicated soaps have extensive studies on their antimicrobial activities, existing research on herbal soaps predominantly focuses on plant extracts rather than finished soap formulations, leaving a gap in understanding the true efficacy of herbal-based soaps against clinically relevant pathogens. Additionally, variations in preparation methods and the absence of standardised testing protocols complicate the comparison and validation of results across different studies. The study aims to evaluate the antimicrobial efficacy of herbal-based soaps using standardised laboratory procedures. A broth microdilution assay was conducted, followed by subculture confirmation to determine the minimum inhibitory concentrations (MICs), minimum bactericidal (MBC) and fungicidal (MFC) concentrations. Time-kill assay assessed concentration- and time-dependent effects at MBC/MFC levels through serial dilutions and colony-forming units (CFU) enumeration. The MIC and MBC values for Staphylococcus aureus were 53.33 mg/mL and 80 mg/mL, respectively, with time-kill analysis showing bacterial reduction within 30 s and complete elimination by 3 min. Candida albicans exhibited higher MIC and MFC values of 120 mg/mL, with complete inhibition observed at 4 min. Overall, the low MIC values and rapid microbial inhibition highlight the influence of soap concentration and exposure time on its potency. Overall, this investigation not only demonstrated the microbial reduction capability of the herbal soap but also contributed valuable methodological insights. By providing a useful testing framework, it paves the way for ongoing improvements in product formulation and ensures that consistent efficacy standards can be maintained. This contributes to the broader field of natural product development and supports the increasing interest in plant-based antimicrobial agents as safe and effective alternatives to conventional chemicals.

Effect of High-Energy Ball-Milling Duration on the Physicochemical Properties of Chemically Treated MD2 Pineapple Peel Cellulose

2026-09-23T12:03:31+00:00

MD2 pineapple peel represents an abundant agro-waste rich in cellulose but remains underutilised in high-value applications. This study investigates the influence of ball-milling duration on the physicochemical properties of cellulose extracted from MD2 pineapple peel. The peel was chemically purified via alkaline delignification, bleaching, drying and high-energy ball milling for 0, 60, 90, and 120 min. The extraction yield of 10.6% (dry basis) was obtained using fresh peel with an initial moisture content of 83.4 ± 1.1%. FTIR confirmed removal of lignin and hemicellulose, while XRD showed diffraction patterns consistent with cellulose I, with crystallinity peaking at 60 min before declining with prolonged milling. SEM revealed initial fibre fragmentation and size reduction at 60 min, followed by increased particle agglomeration with prolonged milling. TGA indicated that milled-extracted cellulose exhibited high thermal stability, comparable to that of commercial microcrystalline cellulose. Among the milling durations investigated, 60 min produced the highest measured crystallinity index (66.67%) and the smallest apparent Feret diameter (193.3 ± 46.6 μm) among the extracted cellulose samples, while thermal stability remained comparable among the milled samples (T_max= 354–355°C). These results highlight the influence of milling duration in refining processing conditions for the effective valorisation of pineapple peel into functional cellulose.

Presence and Effects of Papaya Ringspot Virus (PRSV) on the Leaves of Carica papaya Linn.

2026-09-23T12:03:30+00:00

Papaya (Carica papaya Linn.) is an economically important tropical fruit crop and a major fruit export commodity in Malaysia. However, its production is severely threatened by Papaya ringspot virus (PRSV), which causes significant yield losses in papaya cultivation. Therefore, this study aimed to identify the presence of PRSV and to characterise leaf anatomy and the anatomical adaptations that occur as a result of PRSV infection. Virus identification was performed using reverse-transcription polymerase chain reaction (RT-PCR) targeting the PRSV coat protein plus partial nuclear inclusion b (Nib) region. Anatomical analyses were conducted under a light microscope, including cross-sections of the lamina, midrib, petiole, and leaf margin, as well as epidermal peeling of the adaxial and abaxial surfaces. Micromorphological studies involved examination of the abaxial and adaxial leaf epidermis using a scanning electron microscope. The results showed that PRSV infection induced several anatomical adaptations, including a reduction in chlorophyll content, an increase in the thickness of chlorenchyma and collenchyma cell layers, alterations in vascular tissues, changes in the external morphology of the leaf margin and midrib, and an increase in crystal density. Moreover, changes were observed in the presence and type of epicuticular wax, cuticular ornamentation, anticline wall patterns of the adaxial epidermis, as well as a reduction in stomatal index and stomatal size. These findings demonstrate that PRSV infection causes significant modifications in leaf anatomical characteristics as part of the plant’s response to viral attack. The anatomical indicators identified in this study may serve as valuable references for early detection and management of viral infections in plants.

Pengenalpastian dan Pencirian Gen Mengekod Protein Pengangkut Feromon, Ste6 Kulat Ganoderma boninense

2026-09-25T08:38:39+00:00

Oil palm (Elaeis guineensis Jacq.) is a major crop commodity in Malaysia and Indonesia, contributing more than 80% of global palm oil production; however, its productivity is threatened by basal stem rot (BSR) disease caused primarily by Ganoderma boninense, which results in economic losses of up to USD 500 million annually. Genetic variation in this fungus, generated through sexual mating processes, produces strains with differing levels of virulence, thereby complicating disease management strategies. In pathogenic fungi, mating genes act as key virulence factors as they regulate morphogenesis, proliferation, and the transition from a saprophytic to a pathogenic lifestyle. Although the tetrapolar mating system of G. boninense, involving the matA and matB mating-type loci, has been identified, the pheromone transporter protein Ste6—an essential component of this system—has not yet been reported. This study aimed to identify and characterize the Ste6 protein in G. boninense using bioinformatic approaches and gene expression analysis. The findings revealed that the Ste6 amino acid sequence is highly conserved within the Ganoderma genus, with sequence identity exceeding 90%, and gene expression analysis confirmed its involvement in the mating system. Overall, this study provides the first report of the Ste6 protein in G. boninense. Although its high level of conservation suggests that Ste6 may not be a suitable target for antifungal compound development, the discovery highlights its strong potential as a biomarker for species identification of G. boninense, offering a faster and more accurate alternative to internal transcribed spacer (ITS)-based sequencing.

Root Tensile Behaviour and Soil Shear Reinforcement of Tropical Shrubs for Soil Bioengineering Applications

2026-09-25T08:38:37+00:00

The use of tropical shrub species in soil bioengineering has gained increasing attention for slope stabilization; however, information on their biomechanical contribution to tropical residual soils remains limited. This study evaluated the root tensile behaviour, soil shear reinforcement, and root–soil micromorphology of Strobilanthes crispa (SC) and Pseuderanthemum carruthersii (PC) cultivated for six months. Root tensile tests were conducted across diameter classes, while unconsolidated-undrained (UU) triaxial tests and Field Emission Scanning Electron Microscopy (FESEM) were used to assess soil reinforcement mechanisms and root–soil interactions. Root tensile strength followed an inverse power-law relationship, ranging from 0.72–9.79 MPa for PC and 0.51–21.67 MPa for SC, highlighting the importance of fine roots. In triaxial testing, root-permeated soils exhibited significantly higher maximum shear stress (τ_max) than non-vegetated soil. At 200 kPa confining pressure, SC recorded the highest τ_max (248.9 ± 16.67 kPa), followed by PC (231.1 ± 0.01 kPa), and control soil (214.8 ± 2.30 kPa). Strong linear relationships between τ_max and confining pressure were observed (R² = 0.9689-0.9943), accompanied by slight reductions in apparent cohesion (100.30 kPa in the control soil, 91.80 kPa in SC, and 90.05 kPa in PC) and substantial increases in internal friction angle (25.52°, 35.86°, and 34.24°, respectively). FESEM analysis revealed denser aggregate packing, finer root fibrils (~232.9 µm) and stronger root-soil interlocking in SC than in PC with larger fibrils (~251.9 µm). These findings demonstrate that both species, particularly Strobilanthes crispa, possess favourable biomechanical characteristics for sustainable shallow slope stabilization in tropical environments.

Comparative Effects of ALA and EPA on the Conformational Stability of Wild-Type and Mutant SARS-CoV-2 Spike-ACE2

2026-09-27T09:06:24+00:00

The evolution of SARS-CoV-2 variants through mutations in the Spike (S) protein in the receptor-binding domain (RBD) significantly influences its binding affinity to human Angiotensin-Converting Enzyme 2 (ACE2). While polyunsaturated fatty acids (PUFAs), specifically alpha-linolenic acid (ALA) and eicosapentaenoic acid (EPA), are recognised for their anti-inflammatory properties, their molecular impact on the structural stability and mechanics of the S-ACE2 complex is less understood. This study utilised 100 ns molecular dynamics (MD) simulations to evaluate the modulatory effects of ALA and EPA on both wild-type and mutant S-ACE2 complexes in the presence of the 13 most frequently occurring S mutations in Malaysian patients from August 2023 to January 2024. Analyses of Root Mean Square Deviation (RMSD) and Radius of Gyration (Rg) indicated that the mutant S-ACE2 naturally adopts a more compact yet structurally divergent conformation compared to the wild-type. PUFA binding alters these dynamics; notably, EPA stabilised the mutant S-ACE2 into a conformation resembling the wild-type and markedly decreased per-residue fluctuations (RMSF) in critical interface regions (residues 474–486). Principal Component Analysis (PCA) confirmed that EPA binding restricts the conformational space of both mutant and wild-type S-ACE2. Snapshots at different time points show that the longer carbon chain of EPA binds R408S in mutated S-ACE2, underlying the stable conformation of the complex, whereas ALA, with a shorter carbon chain, does not do so. These findings suggest that ALA and EPA function as structural modulators of the S-ACE2 binding affinities, potentially interfering with the conformational stability required for optimal viral-host interaction. This research provides a molecular basis for the variable impacts of dietary PUFAs to act as structural inhibitors against evolving SARS-CoV-2 variants by disrupting the binding mechanics of the S-protein.

Molecular Detection of Taenia taeniaeformis in Wild Rats and Its Definitive Hosts from Pulau Pinang, Malaysia

2026-09-27T09:06:22+00:00

Taenia taeniaeformis is a zoonotic tapeworm affecting cats and dogs as definitive hosts, rodents as intermediate hosts, and occasionally humans as accidental hosts. Infected animals may suffer from weight loss, vomiting, malnutrition, and intestinal obstruction, while their presence in urban settings raises public health concerns. This study aims to determine the occurrence and identification of T. taeniaeformis in wild rats, domestic cats, and dogs in several selected locations in Pulau Pinang, Malaysia. From October 2022 to March 2023, 146 wild rats were trapped on both the mainland and the island of Pulau Pinang, and faecal samples from cats and dogs were collected from public spaces, veterinary clinics, and shelters. A total of 162 faecal samples (107 cats, 55 dogs) were examined using Sheather’s sugar flotation method. Only one cat (0.9%) tested positive for cyclophyllidean eggs, while none were detected in dogs. For molecular confirmation, 44 randomly selected samples (30 cats, 14 dogs) were examined, which yielded 20 (10 cats, 10 dogs) clear PCR bands, which were subjected to sequencing. Additionally, one cat faecal sample from Cat Beach Sanctuary that was examined positive for cyclophyllidean eggs was also sequenced. However, the resulting sequence did not match T. taeniaeformis sequences in the NCBI database. Based on morphological characterization, all the recovered larvae showed features consistent with T. taeniaeformis. Thus, three randomly selected larval isolates obtained from rats underwent molecular analysis. It was found that one sample (OR790451) yielded a clear PCR band, which showed high similarity with registered T. taeniaeformis sequences. Phylogenetic analysis of 12S rDNA revealed that this isolate clustered closely with a Chinese isolate from the leopard cat (Prionailurus bengalensis, ON055368), confirming its identity. The findings highlight wild rats as important reservoirs and emphasize the need for continued surveillance in companion animals.

Safety Evaluation of Barleria lupulina Lindl. Leaf Extract via Toxicity Assessment and Analysis of Chemical Constituents

2026-09-28T14:14:36+00:00

Barleria lupulina Lindl. is a perennial herb commonly referred to as the hophead Philippine violet and “Penawar Seribu Bisa” or “Bisa Ular” in Malaysia. It belongs to the Acanthaceae family and holds a significant place in the traditional pharmacopeia of indigenous communities in Malaysia. This study aims to assess the safety profile of the leaf extract, focusing on its elemental composition and potential cytotoxicity. The concentrations of heavy metals were quantified using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS), while cytotoxicity was assessed through the Brine Shrimp Lethality Assay (BSLA). Additionally, Ultra-High-Performance Liquid Chromatography coupled with Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-QTOF/MS) was utilized to characterize the chemical constituents of the extract. Results indicate that the heavy metal concentrations in B. lupulina leaf are within the permissible limits set by the Codex Alimentarius, suggesting they are safe for consumption. The BSLA assessment demonstrated no significant mortality in brine shrimp, even at the highest extract concentration of 1000 μg/mL, further supporting the safety of the extract. UHPLC-QTOF/MS analysis revealed the presence of several compounds—paeonolide, sibiricaxanthone B, scroneoside A, roseoside, and cornuside II—reported for the first time in this species.

Fungal Metabolites as Multi-Target Modulators of Alzheimer’s Disease: An Integrated In Silico Docking and Molecular Dynamics Approach

2026-09-28T18:01:14+00:00

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder with limited therapeutic options offering only symptomatic relief. Natural fungal metabolites have recently gained attention for their neuroprotective potential. In this study, an extensive library of 18,349 fungal compounds was retrieved from the NPAtlas database and systematically screened for drug-likeness, ADMET properties, and toxicity profiles. Successive filtering reduced the dataset to 99 drug-like molecules, of which five candidates—Sonomolide A, Steperoxide A, RP-1551-M2, Illudin J, and Pericoannosin F—were selected for detailed analysis. These compounds demonstrated favourable physicochemical properties, compliance with Lipinski’s Rule of Five, high gastrointestinal absorption, blood–brain barrier permeability, and acceptable toxicity margins (Class 6). Molecular docking against five AD-related targets (AChE, CBNR2, CHRM1, CHRM4, & D1B dopamine receptor) revealed strong binding affinities, with Steperoxide A showing the highest affinity for AChE (−8.0 kcaL/moL) and CBNR2 (−9.0 kcaL/moL), while RP-1551-M2 displayed consistent interactions with muscarinic and dopamine receptors. MD simulations confirmed stable protein–ligand interactions, with RMSD values ranging from 0.10–1.75 nm and hydrogen-bond counts between 47–252 throughout the trajectory. Stable radius of gyration values (2.74–3.95 nm) further indicated the maintenance of structural compactness and binding integrity, and the results also showed good stability and favourable interaction profiles. Collectively, these findings highlight fungal metabolites—particularly Steperoxide A and RP-1551-M2—as promising multi-target neuroprotective scaffolds with potential therapeutic relevance in AD.

In Silico Profiling of Small Open Reading Frames in hsdM-Deficient Group A Streptococcus and Their Predicted Role in Membrane Adaptation

2026-09-28T14:26:26+00:00

Small open reading frames (sORFs) are emerging as key players in bacterial gene regulation, yet their roles remain underexplored in many pathogens. This study investigates the regulatory landscape of sORFs in Group A Streptococcus (GAS) by integrating transcriptomic data and weighted gene co-expression network analysis (WGCNA). We focused on the functional consequences of inactivating the Type 1 Restriction-Modification (RM) system, specifically the hsdM subunit, which is a known barrier to genetic transformation in GAS. By reanalysing the transcriptomic profile of the hyper-transformable 854ΔhsdM strain, we identified 28 sORFs localised within significantly altered co-expression modules. Notably, sORF_00369 (associated with fatty acid biosynthesis) and sORF_00282 (linked to ABC transport) were significantly downregulated. These findings suggest that the inactivation of the RM system, while improving transformation efficiency, may trigger a targeted shift in sORF-mediated metabolism and membrane integrity. Our results propose that sORFs are candidate regulators within the epigenetically mediated regulatory networks of GAS, providing new insights into how genetic recalcitrance and metabolic fitness might be linked. This in silico study lays a theoretical foundation for potentially targeting sORFs in antimicrobial strategy development and optimising GAS as a chassis for synthetic biology.

Molecular Characterization of Indigenous Potassium-Solubilizing Bacteria from Rice Rhizosphere in Lima Puluh Kota Regency

2026-09-28T18:15:06+00:00

Potassium (K) is one of the macronutrients essential for plant growth; however, most soil K exists in insoluble mineral forms that are not readily available for plant uptake. The use of potassium-solubilizing bacteria (KSB) offers a sustainable approach to enhance K bioavailability in agricultural systems. This study aimed to isolate, characterize, and identify KSB from the rhizosphere of seven local rice varieties cultivated in Lima Puluh Kota Regency, West Sumatra, Indonesia. Total culturable heterotrophic bacterial populations were enumerated using nutrient agar, and one morphologically distinct colony from each sample was purified for further analysis. K-solubilizing activity was evaluated on Aleksandrov agar by measuring the dissolution index (DI) and in liquid Aleksandrov medium supplemented with K-feldspar powder. All seven isolates formed clear halo zones on solid medium. The highest DI value was observed in isolate KSP (2.2), followed by KKP and KJG, while KRK exhibited the lowest DI. In liquid medium, soluble K release ranged from 0.95 to 1.95 mg L⁻¹, with KSP demonstrating the highest solubilization capacity. Statistical analysis revealed significant differences among isolates (p<0.05). Molecular identification based on 16S rRNA gene sequencing indicated that the isolates belonged to Lysinibacillus xylanilyticus, Bacillus velezensis, Acinetobacter johnsonii, Pseudarthrobacter phenanthrenivorans, Klebsiella aerogenes, Paenarthrobacter nicotinovorans, and Enterobacter cloacae. These findings demonstrate the diversity and functional potential of indigenous KSB associated with rice rhizosphere soils and support their prospective application as biofertilizer candidates for sustainable rice production.

Safety Evaluation of Hot Water and Ethanol Extracts of Crescentia cujete L. Bark in Zebrafish Embryo Model for Potential Functional Ingredient Applications

2026-09-29T17:07:12+00:00

The exploration of natural plant sources for functional ingredients has gained significant attention in food science, with emphasis on bioactive compounds that can enhance human health. Crescentia cujete L. (calabash tree) is traditionally valued in folk medicine, yet the safety of its bark extracts for functional food applications remains underexplored. This study evaluated the safety performance of hot water and ethanol extracts of C. cujete bark using the zebrafish embryo toxicity (ZFET) model, an established in vivo tool for early toxicological screening. Fertilised zebrafish embryos were exposed to graded concentrations of each extract from 0 to 120 hr post-fertilisation (hpf), and endpoints including survival, hatching, heart rate, and morphological development were monitored. The hot water extract demonstrated a significantly higher median lethal concentration (LC₅₀) of 9.69 mg/mL, while the ethanol extract exhibited a markedly lower LC₅₀value of 0.42 mg/mL (p<0.001), suggesting higher relative safety of the aqueous preparation. Dose-dependent reductions in survival and hatching rates were observed, with ethanol extracts producing more severe developmental delays and cardiac abnormalities. Heart rate disturbances, pericardial edema, and axial malformations were most prominent at concentrations near the ethanol LC₅₀value, whereas the hot water extract induced only mild effects at higher concentrations. No significant changes in heart rate were observed with the hot-water extract, whereas significant reductions were observed at higher concentrations of the 50% ethanol extract (p<0.05). These findings indicate that the extraction solvent significantly influences the safety profile of C. cujete bark. The hot water extract demonstrated lower developmental toxicity in the zebrafish embryo model and warrants further investigation through phytochemical characterisation and mammalian toxicity studies. By establishing the first zebrafish-based safety evaluation of C. cujete bark, this study highlights the importance of integrating toxicological validation in the development of novel plant-derived functional ingredients.

Assessing the Suitability of Reference Genes for Normalising Gene Expression in Ganoderma boninense-Infected Oil Palm

2026-09-29T17:07:14+00:00

Oil palm is a major agricultural commodity in Southeast Asia, particularly in Malaysia and Indonesia. Its productivity, however, is being jeopardised by climate change and disease outbreaks, particularly basal stem rot (BSR) caused by Ganoderma boninense. Monoculture practices exacerbate disease spread, calling for innovative and sustainable solutions. Beyond current integrated plant management (IPM) practices, developing disease-tolerant plant material could enhance disease management. For that purpose, efforts have been initiated to identify potential genetic markers associated with G. boninense tolerance; include in vitro screening (IVS) using oil palm clonal material is employed, where artificial infection is conducted under a controlled environment. However, the reliability of such studies depend on accurate quantification of gene expression, which requires the use of reliable reference genes (RGs). Quantitative PCR (qPCR) is a powerful tool for measuring gene expression levels; however, reliable normalisation depends on RGs that maintain consistent expression across tissues and experimental conditions. Consistent RG expression is essential for minimising technical and biological variation, ensuring that observed changes in target gene expression reflect true biological responses rather than variation arising from the normalisation process. Therefore, this project evaluated ten potential RGs selected from the literature for gene expression analysis in young oil palm ramets under controlled G. boninense-infected conditions. Among the ten RGs evaluated, ubi and fbox were consistently identified as the most stable and reliable RGs for in vitro expression analysis under controlled disease-stressed conditions, based on geNorm, NormFinder, BestKeeper, and RefFinder analyses. In contrast, gapdh was unsuitable as an RG under these conditions. The RGs identified and validated in this study are anticipated to enhance the accuracy of gene expression analysis in G. boninense-infected oil palm ramets under controlled in vitro conditions. This approach will be instrumental in identifying potential defence-related genes associated with G. boninense tolerance through comprehensive gene expression profiling.