Malaysian Applied Biology

Enhancing Jackfruit’s Bioactive Properties Through SCOBY Fermentation: Implications For Cosmeceuticals

Norhazniza Aziz, Koh Soo Peng, Norefrina Shafinaz Md Nor — Thu, 27 Mar 2025 00:00:00 +0000

This study investigated the bio-processing technique of fermenting jackfruit pulp (JP) and leaves (JL) using a symbiotic culture of bacteria and yeast (SCOBY) to enhance their bioactive properties. To assess the nutritional value of the jackfruit extracts, the phenolic and organic acid compositions were determined. The extracts were also evaluated for their anti-inflammatory properties by measuring their ability to suppress the production of nitric oxide (NO) in response to bacterial lipopolysaccharide (LPS) stimulation in RAW 264.7 macrophage cell line. Furthermore, the inhibitory effects of the extracts on elastase and tyrosinase, enzymes associated with skin aging, were assessed. The fermentation process led to increased phenolic content. This included vitexin, salicylic acid, and benzoic acid. Acetic acid was the most abundant organic acid detected after fermentation, with concentrations ranging from 16.0 to 16.1 mg/mL. Additionally, the fermented extracts exhibited elevated levels of other beneficial organic acids such as citric and quinic acid. The study demonstrated significant reductions in nitrite formation in LPS-stimulated RAW 264.7 cells treated with jackfruit extracts. This finding suggests that the fermented extracts can effectively suppress NO production in a concentration-dependent manner. Among the fermented extracts, JL exhibited the highest inhibition of NO production at a concentration of 19.5%, resulting in a 42.23% reduction. Moreover, fermentation enhanced the inhibitory effects of the jackfruit extracts on elastase and tyrosinase, with inhibition rates ranging from 82.3% to 95.4%. Overall, the findings suggest that fermented jackfruit exhibits increased levels of phytochemical compounds and holds promise as a natural and beneficial ingredient in cosmeceutical products, offering anti-inflammatory and skin-aging benefits.

An Effective Disinfection Protocol for The Development of Tissue Culture Mango Plant (Mangifera indica L.) cv. Harumanis

Zun Yip Lee, Amir Hamzah Ahmad Ghazali, Bee Lynn Chew, Sreeramanan Subramaniam — Thu, 27 Mar 2025 00:00:00 +0000

Harumanis mango is one of the highest demand mango cultivars in Malaysia due to its exceptional sweetness and fabulous fragrance. However, the production of this mango cannot meet the market demand due to limited grafting activity and the limited number of seeds (only one harvest season per year). These make vegetative and non-vegetative propagation of Harumanis time and labour intensive. Micropropagation using tissue culture techniques is a reliable and effective alternative for mass in vitro propagation of Harumanis at a consistent and faster rate, producing clones of the mother plants. Nevertheless, deep-seated contaminants are the major problem faced in the micropropagation of this plant. The objective of the present study is to develop an effective disinfection protocol for Mangifera indica cv. Harumanis. A sterility rate of 87.5% was achieved by spraying the mother plants with 0.5 mL/L azoxystrobin two days before the experiment (pre-treatment) and surface sterilising nodal segments at the immature green leaf stage with 40% Clorox® for 20 min, 0.5 g/L benomyl for 1 hr and 300 mg/L cefotaxime for 5 min. The transverse thin layer (tTCL) technique was applied to the nodal segments and cultures were maintained at WPM with 0.5 g/L benomyl and 300 mg/L cefotaxime. Endophytic bacteria and fungi were observed in the axillary buds using scanning electron microscopy (SEM). Both histological and SEM analysis showed that the xylem vessels of the nodal segments at these two ends of the stem tended to have lower numbers of endophytic bacteria. This suggests that the nodal segments at the upper and middle ends of the stem are the best starting material for future experiments. The present results show the importance of pre-treatment and culture maintenance in reducing endophytic contamination. Furthermore, the results showed the potential of tTCL-treated Harumanis cultures in minimising the contamination rate.

Evaluation of Morinda citrifolia Leaf Extract Against Phytophthora palmivora in Controlling Stem Canker on Durian (Durio zibethinus)

Nor Dalila Nor Danial , Norhayu Asib , Tosiah Sadi , Siti Izera Ismail — Thu, 27 Mar 2025 00:00:00 +0000

Durian is an economically important crop in Malaysia and has been identified as a new source of agricultural wealth valued at USD 1.58 billion. However, Phytophthora palmivora has been reported in all areas where durian has been planted and continues to pose a significant challenge for durian growers. The crop losses and control costs were estimated at 11.7% in 2020. Durian farmers have applied oomycete fungicides for more than 20 years to control stem canker, and extensive use of fungicides has resulted in the development of resistant Phytophthora isolates. This study aimed to evaluate four different leaf crude extracts of mengkudu (Morinda citrifolia) extracted using methanol, acetone, ethyl acetate, and hexane solvents against P. palmivora. GC-MS analysis of M. citrifolia leaves crude extract revealed the presence of squalene. In-vitro screening of the M. citrifolia plant extracts, the crude extracts of acetone and ethyl acetate revealed effective concentration 50% (EC₅₀) values of 36.115 and 38.095 mg/mL, respectively. PDA supplemented with acetone solvent at 60 mg/mL altered the morphology and inhibited the mycelial growth of P. palmivora. In an in-vivo bioassay screening level, treated plants of M. citrifolia extract of ethyl acetate 76 mg/mL had a lesion growth of 30.24 mm on the stem. This treatment was not significantly different from the control positive treatment applied with Ridomil fungicide. Morinda citrifolia leaf extracts using methanol, acetone, and ethyl acetate revealed it could be used as an alternative biocontrol agent for foliar spraying to reduce disease severity in durian seedlings against P. palmivora.

Improving The Sporulation of Eimeria tenella Oocysts Purified From Chicken Faeces

Thu, 27 Mar 2025 00:00:00 +0000

Coccidiosis is a major and recurrent intestinal disease in the chicken industry caused mainly by Eimeria tenella and it is the best-known and most economically significant species in poultry. Oocyst purification is a common and critically important method used for the study of Eimeria species. Various methods have been used for the purification of coccidia oocysts. However, no study has outlined the most effective method for increasing the sporulation of Eimeria tenella oocysts. Thus, the study compared and validated the applications of the three common oocyst sporulation methods, viz: (i) forced aeration without a shaker, (ii) forced aeration in combination with a shaker, and (iii) magneto base without forced aeration. A total of 9 individual 21-day-old broiler chickens were inoculated individually with a single 1 mL dose containing 2000 Eimeria tenella oocysts via oral gavage. Water and food were provided ad libitum throughout the experimental period. Pooled group faecal samples were collected on the 7 day post-infection. The average (range) number of sporulated oocysts counts for the forced aeration without shaking was 2250 (1000-2000), forced aeration in combination with shaking was 2000 (1000-1500), and the magneto base without forced aeration was 5500 (2000-3500). The results from the study indicated that the magneto base without forced aeration was the most effective method for enhancing the sporulation of Eimeria tenella oocysts.

Antifungal Activity of Aegle marmelos (L.) Corrêa and Cassia alata L. Extracts Against Candida krusei and Candida parapsilosis

Thu, 27 Mar 2025 00:00:00 +0000

Candidiasis is a fungal infection caused by several Candida species, including Candida krusei and Candida parapsilosis. It poses a significant threat to human health, particularly in immunocompromised individuals and those with underlying medical conditions. The rapid development of microbial resistance against antibiotics and the emergence of new strains are burdens to the global health community. Medicinal plants could be an excellent source for their potential antimicrobial effects. Therefore, this study aims to assess the antifungal action of Aegle marmelos and Cassia alata extracts on the growth and morphological changes of Candida krusei and Candida parapsilosis. The pathogens were cultured on media containing each extract individually. Itraconazole was used as a positive control, whereas dimethyl sulfoxide (DMSO) was used as a negative control. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by the microdilution method. All samples were processed for microscopy observations using a scanning electron microscope (SEM). The MIC of methanolic extract indicated that the leaf and unripe fruit of A. marmelos and C. alata against C. parapsilosis and C. krusei were similar at 75 mg/mL. Results of the MFC showed both pathogens were killed at a concentration of 150 mg/mL for both plant extracts. The MIC and MFC results of the methanolic extract indicated that the unripe fruit of A. marmelos exhibited fungistatic activity, while the leaves of both C. alata and A. marmelos demonstrated both fungicidal and fungistatic effects against C. parapsilosis and C. krusei. SEM observations showed significant changes in the morphology of C. krusei and C. parapsilosis compared to the control, in which the fungi were in normal form. The findings indicate the potential use of A. marmelos and C. alata methanolic extract as antifungal agents for candidiasis treatment. The extracts showed comparable inhibitory action to the commercial fungicide itraconazole.

Formulation of Anti-Acne Gel Containing Citrus aurantifolia (Christm.) and Aloe barbadensis (L.) Extracts and Evaluating The Impact of High-Pressure and Microwave Processing

Thu, 27 Mar 2025 00:00:00 +0000

Acne is one of the most prevailing skin disorders caused by bacteria, dead skin cells, and oil clogging of hair follicles. In this study, a polyherbal anti-acne gel containing C. aurantifolia and A. barbadensis extracts was developed as a cosmeceutical. Major bioactive fractions in the plant extracts were evaluated by gas chromatography-mass spectrometry (GCMS) analysis. Moreover, C. aurantifolia depicted higher total phenolic, flavonoids, tannins, and ascorbic acid than A. barbadensis extract. The anti-acne gel was prepared by adding 1% of both plant extracts and evaluated for organoleptic properties (color, odor, homogeneity, consistency, washability), spreadability, viscosity, extrudability, pH, and drug contents and compared with a commercial herbal formulation (NuTeen^®). The developed gels depicted greater inhibition of Staphylococcus aureus than the commercial formulation with a growth inhibition diameter of 2.40 mm. In-vitro permeation of plant extracts from a gel into phosphate buffer was found at 27.4% after 2.5 hr, and the release behavior was best explained by the Higuchi model (R²=0.97). Finally, for the possible replacement of paraben (synthetic preservative) from the gel, high-pressure processing (600 MPa, 120 s), and microwave pasteurization (700 W, 80 s) were adopted and the stability of gels was evaluated after 4 weeks, and found comparable to their paraben-containing normal counterpart.

Characterization of Dietary Fibers from Corn Cobs as A Potential Functional Ingredient in Muffin

Thu, 27 Mar 2025 00:00:00 +0000

Dietary fiber (DF) has gained significant attention due to its potential health benefits. Corn cobs, a by-product of corn that is rich in minerals and low in protein, represent a valuable source of DF. This study investigated the fiber content of corn cobs and evaluated its potential use in bakery products to provide enhanced nutritional options. The physicochemical properties of the extracted fiber, including Fourier transform infrared spectroscopy, water-holding capacity (WHC), oil-holding capacity (OHC), swelling capacity (SC), glucose adsorption capacity (GAC), and cholesterol adsorption capacity (CAC), were assessed. Three extraction methods were employed: water, acid, and alkali. Water extraction yielded the highest total DF at 98.12% (p<0.05) and the highest insoluble DF at 69.40%, excelling in OHC, SC, and CAC. Alkali-extracted fiber demonstrated superior WHC, while acid-extracted fiber showed the highest GAC. Muffins enriched with fiber extracted using these methods were compared to the control group (without fiber), revealing no significant differences in texture, except for cohesiveness. In conclusion, water extraction is the most advantageous method, providing the highest yields, enhanced safety, and a muffin texture comparable to those made with fiber from acid and alkali extraction without compromising quality.

In vitro Regeneration and ISSR-Based Genetic Fidelity Evaluation of Stevia rebaudiana

Zarina Zainuddin, Zannat Urbi, Nurul Madiha Ab Halim — Thu, 27 Mar 2025 00:00:00 +0000

Steviol glycoside, which is a natural sweetener extracted from Stevia rebaudiana, is globally recognized. For consumers, this compound is widely utilized by diabetic patients and demonstrates numerous therapeutic effects. However, the escalating demand for this natural sweetener and medicinal herb impacts the availability of stevia. Conventional propagation methods, such as seed and stem cutting, frequently result in low germination rates. To address these limitations, the present research explores the potential of in vitro clonal propagation to ensure a consistent supply of planting material. Therefore, the objective of this study was to develop an efficient protocol for tissue culture of S. rebaudiana accession MS007. The highest regeneration frequency (85.19% & 86.67% for shoot tips & nodes, respectively) and maximum shoot number (14.30 & 12.77 shoots/explant, respectively) were observed on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 6-benzylaminopurine (BAP). Half-strength MS medium supplemented with 0.5 mg/L indole-3-butyric acid (IBA) was the optimal medium for rooting, exhibiting the highest rooting percentage, root number, and root length. Subsequently, the plantlets were acclimatised in plastic cups containing peat moss, and it was observed that 86.67% of plantlets survived when transplanted to the field. The outcome of inter-simple sequence repeat (ISSR) analysis using 38 ISSR primers confirmed the genetic fidelity between the in vitro regenerated plants and the mother plant. This study successfully developed an in vitro propagation technique for stevia to produce true-to-type clonal plants. The obtained results can be used to mass-produce stevia accession MS007 to meet market demand.

Natural Deep Eutectic Solvents vs. Conventional Solvents: Effects on Crude Yield, Mangiferin Content, Antioxidant Activity, and Toxicity in Mangifera pajang Kosterm. Fruit Extracts

Sat, 29 Mar 2025 00:00:00 +0000

Mangifera pajang Kosterm. fruits, commonly known as 'Bambangan,' are rich in natural antioxidants due to their phytochemical constituents and are categorised as an underutilised fruit. This study focused on investigating the effects of different solvents on the extraction yield (EY), mangiferin content (MC), total flavonoids (TF), total phenolics (TP), antioxidant activity, and toxicity of M. pajang fruit extracts (MPFE). The selected solvents included natural deep eutectic solvents (NADES), water, methanol, and ethanol. The extraction of MPFE was performed using ultrasound-assisted extraction with an ultrasonic probe. Antioxidant activities were evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric-reducing antioxidant power (FRAP) assays. Toxicological assessment was conducted using the brine shrimp lethality assay (BSLA) to determine LC₅₀ values. Overall, NADES proved to be the most efficient solvent for extraction, yielding the highest EY (37.09 ± 2.34%), MC (0.032 ± 0.000 mg/g), TF (0.80 ± 0.01 mg RE/g dry extract), TP (14.94 ± 1.74 mg GAE/g dry extract), and exhibits potent antioxidant activity as measured by DPPH (82.38 ± 0.24%), ABTS (86.37 ± 0.03%), and FRAP (304.57 ± 5.24 mg TE/g dry extract). Moreover, NADES demonstrated non-toxicity in the BSLA (LC₅₀ = 1988.37 µg/mL) of MPFE. These findings suggest that NADES is a suitable solvent for exploring the medicinal potential of M. pajang fruits and their application in therapeutic development.

Evaluation of Small Extracellular Vesicles Isolation Methods from Human Serum for Downstream miRNA Profiling

Thu, 27 Mar 2025 00:00:00 +0000

Exosomes are a type of extracellular vesicles that carry distinct profiles of biomolecules such as lipids, proteins, DNAs, and RNAs. Despite many years of research, there is still a lack in standardized method to isolate exosomes from clinical samples for their downstream applications. Thus, this study compared three different methods, which are the differential ultracentrifugation (DUC), polyethylene glycol-based precipitation (PEG), and a combination of both (PEG+UC) to isolate exosomes from human serum. The isolated exosomes were evaluated by their size distribution, recovered particle concentration, particle to protein ratio, exosomal marker expression, and miRNA recovery. Our results indicated that all three methods successfully isolated exosomes, however, with varying yield and purity. In particular, PEG+UC produced exosomes of both high yield and high purity, DUC produced exosomes of both low yield and low purity, whereas PEG produced exosomes of high yield but low purity. Using miR-30d-5p and let-7i-5p as selected targets, our qPCR results indicated significant differences in terms of exosomal miRNA recovery between all three methods. Overall, the PEG+UC method appeared to be a less labour-intensive alternative that can isolate exosomes of both high yield and high purity from human serum without compromising the yield of miRNAs.

Evaluation of Control Methods for Acusta tourannensis (Souleyet,1842) (Gastropoda: Camaenidae) Infesting Dragon Fruit (Selenicereus spp.) in Vietnam

Thu, 27 Mar 2025 00:00:00 +0000

Dragon fruit (Selenicereus spp.) cultivation in Vietnam faces severe losses due to Acusta tourannensis infestations, impacting yield and fruit quality. This study evaluated the efficacy of various control methods through field and greenhouse experiments in Binh Thuan Province, Vietnam, in 2023. Cultural practices, including reduced irrigation, herbicide application, and grass cutting, effectively suppressed snail populations by creating unfavorable conditions. Mechanical and manual removal methods, although useful, were labor-intensive. Bait trapping with dragon fruit parts was highly effective in capturing snails. Biological control using duck release significantly reduced snail densities, though contamination risks remain a concern. Botanical control with Saponin HF achieved mortality rates of 86.7–96.7% within 7–14 days. Chemical control with metaldehyde and niclosamide provided up to 100% mortality but necessitates careful management due to environmental risks. Integrated pest management (IPM) combining cultural, biological, and botanical control methods offers the most sustainable approach for managing A. tourannensis in dragon fruit orchards. Further research should focus on optimizing application rates and timing, developing eco-friendly alternatives such as plant-derived molluscicides, and assessing the long-term ecological impacts of these strategies. Enhancing these approaches will contribute to the effective and sustainable management of A. tourannensis, ensuring the viability of dragon fruit cultivation in Vietnam.

Metabolic Disturbances, Histopathological Analysis, and Virus Detection in Chicken Following Infection with Fowl Adenovirus Serotype 8B

Thu, 27 Mar 2025 00:00:00 +0000

Fowl adenovirus (FAdV) serotype 8b is a causative agent of inclusion body hepatitis (IBH) in chickens with serious economic impact due to huge mortality in commercial farms. To date, analysis of metabolic disturbances caused by FAdV serotype 8b infection in chicken is scanty although FAdV is highly pathogenic in specific pathogen free (SPF) chicken. It is important to highlight the effect of the FAdV infection for better control strategy against the disease. The objective of the study is to determine the metabolic parameters, virus detection, and histopathological analysis of chickens following infection with FAdV serotype 8b. Fifteen-day-old SPF chicks were divided into two groups namely, the infected group and the control group. All chicks in the infected group were inoculated with 1 mL FAdV isolate, UPM1901, via oral route at day 7 of age, while all chicks in the control remained uninoculated. At 0, 3, and 7 days post-inoculation (dpi), blood and serums were collected for clinical biochemistry analysis followed by histological examination and virus detection using polymerase chain reaction (PCR). Marked anemia with an elevation of aspartate-aminotransferase (AST) and gamma-glutamyl transferase (GGT) levels in infected chickens at 3 and 7 dpi with low creatine kinase levels than control group were recorded due to liver damage caused by FAdV infection. Swollen, hemorrhage, and necrosis of the liver were observed in chicken from the infected group at 3 and 7 dpi with evidence of basophilic intranuclear inclusion bodies in degenerated hepatocytes. Liver, kidney, and lymphoid organs were positive for FAdV at 3 and 7 dpi in the infected group. It was concluded that FAdV serotype 8b isolate UPM1901 induces metabolic disturbances and histopathological lesions with detectable viral nucleic acid in the liver and lymphoid organs which necessitates effective therapeutic strategy for liver protection and a better immune response against IBH disease.