Malaysian Applied Biology

Encapsulated Phage Cocktail: The Destined Environmental Biocontrol Agent For Pathogenic Leptospira

Kokxin Fang — 2024-06-30

Leptospirosis is one of the often-neglected fatal zoonotic diseases endemic to most developing countries. The disease transmits mostly through contact of rodent urine contaminated with pathogenic Leptospira in the environment. This review discusses the need for environmental bioremediation of these pathogens and the reasons phage could very well be employed for that purpose. With a few modifications like encapsulation and cocktail formulation, the functionality and stability of phage as the natural predator could easily be heightened. Host specificity, ability to auto-dose and co-evolve along its hosts, effectiveness against biofilms and independence of its production are some of the promising features of a phage. Here we also highlight the interactions and interference among phages in a cocktail, transduction probability, and hypothetic usage of phage lysin in biocontrolling pathogenic Leptospira.

Probiotic Growth Pattern and Physicochemical Evaluation of Water Kefir Fermentation

Phin Yin Sin — 2024-06-30

Probiotics are live-friendly microorganisms that can confer a health benefit on the host if it is consumed in sufficient amounts. Water kefir is a probiotic-rich fermented beverage that contains multi-species of live cultures. Brown sugar and palm sugar were used for water kefir fermentation due to their high sucrose and mineral contents. The objective of this study was to determine the probiotic growth pattern of water kefir and to evaluate the physicochemical parameters, including the pH changes, lactic acid content, reducing sugar content, and total soluble solids. The fermented water kefir was collected at every 6-hour interval, until the end of 72 hours of fermentation. The growth curve was determined by enumerated probiotics on De Man, Rogosa, and Sharpe (MRS) agar, Yeast Extract-Peptone-Dextrose (YPD) agar, and Gluconobacter (GM) agar plates, respectively. MRS, YPD, and GM agar plates were used to enumerate lactic acid bacteria, yeast, and acetic acid bacteria, respectively. The result showed increased probiotic growth as fermentation time increased with different phases observed from the growth curve. The stationary phase of probiotics was recorded at 30-42 h and was recommended as the optimal harvesting point. Besides, longer fermentation time produced lower pH values and lower total soluble solids while higher lactic acid and higher reducing sugars. At the end of fermentation, the concentration of lactic acid and reducing sugars were 2.16 ± 0.09 g/L and 13.66 ± 0.14 mg/mL, respectively. In conclusion, probiotics from water kefir fermentation are suggested to be best harvested between 30-42 hours and can be used for self-consume or downstream processing.

A Severe Clinical Case of Theileriosis with Terminal Wasting in Adult Cattle: A Veterinary Clinical and Pathology Case Report

Faez Firdaus Abdullah Jesse — 2024-06-30

This veterinary clinical and pathology case report describes a severe clinical case of theileriosis in adult cattle with prolonged recumbency and terminal wasting because of complications due to chronic theileriosis in two adult (2-year-old) Jersey-Friesian Cross. The case was referred to the large animal clinic of the University Veterinary Hospital (UVH), Faculty of Veterinary Medicine, Universiti Putra Malaysia (UPM). The animals were presented as recumbent, dull, depressed, hypothermic, dehydrated, and cachexic. There were poor responses of the patients to fluid therapy and non-steroidal anti-inflammatory treatments, and they both died within 24 hrs of clinical presentation. Haematology (complete blood count and serum biochemistry) results revealed haemoconcentration and faecal examinations were negative for liver fluke infestation and blood smear examination was positive for intraerythrocytic piroplasms of Theileria (0.01% parasitaemia). Post-mortem examination further revealed advanced cachexia, generalised serous atrophy of fat, hepatomegaly, hepatic congestion, pulmonary congestion, and pulmonary oedema. Histological findings revealed atrophy of the hepatic cord, hepatic congestion, splenic hemosiderosis, pulmonary oedema and congestion. The clinicopathological findings supported an aetiological diagnosis of a severe clinical case of Theileriosis with terminal wasting because of complications due to chronic Theileriosis infection.

Identification of Endophytic Actinomycetes Isolated From Hopea ferrea and Its Antibacterial Activity Against Cariogenic Bacterium

Nur Raihan Aqilah Mohammad Azmin — 2024-06-30

Dental caries affects around 36% of the world's population and results in the loss of primary teeth in about 530 million youngsters. It is described as the loss of the enamel layer of the tooth by acids generated by the activity of cariogenic bacterium such as Streptococcus mutans on carbohydrates. This study aims to identify bioactive endophytic actinomycetes as an antibacterial agent against S. mutans DSM 20523 and to identify the bioactive endophytic actinomycetes using the 16S rRNA sequencing approach. To determine the percentage zone of inhibition, three endophytes at 10-day-old culture have been streaked onto ISP2. Then, S. mutans was streaked perpendicular to the endophytes. Crude extracts from three endophytes have been prepared by using four different types of media namely Tryptic Soy broth, Kings B, International Streptomyces Project Medium No.1 (ISP1) and Starch Casein broth. These crude extracts proceeded to undergo disk diffusion and MIC assays against S. mutans. All three endophytes from streaked plates with the highest percentage zone of inhibition were identified using the 16S rRNA gene molecular approach for characterisation. Endophytic actinomycetes which were isolated from Hopea ferrea showed potent antibacterial activity against S. mutans. All endophytic actinomycetes isolates from streak plates showed good inhibitory activity against the S. mutans. The highest percentage of inhibition was shown by the PT9-13S2 isolate followed by PT9-8S2 and PT9-13W2. Furthermore, crude extracts of PT9-8S2 and PT9-13W2 (the highest ZOI) from TSB media showed inhibitory activities against S. mutans, from disk diffusion as well as minimum inhibitory assays. From the 16S rRNA molecular approach, endophytic actinomycetes PT9-8S2 was closely related to Streptomyces collinus while PT9-13S2 and PT9-13W2 were closely related to Streptomyces malachitospinus. From this study, endophytic actinomycetes showed a promising source as antibacterial agents against cariogenic bacterium S. mutans which is the main causative agent of dental caries.

Seed Longevity of Soybean (Glycine max L.) Cultivars Under Lightless Storage

Kadapi Muhamad — 2024-06-30

Soybean seeds in tropical countries are prone to rapid deterioration due to environmental condition during storage, leading to a decrease in seed quality and ultimately resulting in low potential yield. Longevity of seed during storage is influenced by genetic and environmental factors such as light. Therefore, this study aimed to determine the effect of light on the vigour of two soybean seeds cultivars (Demas 1 & Devon 1 cvs) stored for three months. The investigation was conducted from February to May 2022 at Seed Technology Laboratory, Faculty of Agriculture, Universitas Padjadjaran. A correlational method was employed to assess the relationship between seed quality of Demas 1 and Devon 1 in light and lightless storage. Furthermore, data were analyzed using correlation regression and t-test to compare the two populations. The results showed that there was a high correlation between the physical and physiological quality of seeds. There were also differences in seed response to light, and this was observed from the moisture content, weight of 100 seeds, germination rate, and simultaneous growth percentage traits. Considering the seed quality traits, Demas 1 had the best vigour value after storage in dark conditions. It is worth noting that both cultivars exhibited signs of deterioration during storage. However, the rate of deterioration was slowed down due to lightless condition.

Salmonella Isolated From Raw Chicken Meats at Selected Slaughterhouses in Peninsular Malaysia; Their Antibiotic Resistance Profiles and Biofilm Formation on Nutrient-Limited Media

Zuraidah Ismail — 2024-06-30

Salmonella is one of the pathogens responsible for foodborne diseases. Antibiotic resistance of Salmonella, particularly multidrug-resistant (MDR) strains have emerged and are becoming more prevalent, which is a very serious issue worldwide. This study sought to determine the antibiotic resistance profiles of Salmonella isolated from raw chicken meats, which were collected at selected slaughterhouses in Peninsular Malaysia and evaluating its biofilm-forming capability on surfaces. Antibiotic resistance of 135 Salmonella isolates against 12 antibiotics were investigated via disk diffusion method. The biofilm-forming ability of the isolates was evaluated by crystal violet staining using two media; a tryptic soy broth (TSB) and a 1/20 TSB with incubation periods of 24 and 48 h at 37 °C. A total of 118 strains of Salmonella showed higher resistance to erythromycin (87.41%), followed by tetracycline (85.19%;); 93 of the isolates (68.88%) were multi-drug resistant. A greater quantity of Salmonella was able to produce biofilm when grown in 1/20-TSB (90.37%) compared to the growth in TSB (88.15%), respectively. The findings in this study showed high prevalence, antibiotic resistance, and the biofilm forming ability of Salmonella strains isolated from raw chicken meats, suggesting that effective measures are required to ensure food safety in the poultry industry.

Phytochemical Evaluation and Anti-angiogenic Activity of Alingatong (Dendrocnide meyeniana Walp.) Root Extracts Using the Chorioallantoic Membrane Assay on Duck Embryo

Ezra Maurel D Torno — 2024-06-30

Dendrocnide meyeniana (Walp.), known as "Alingatong" in the Visayas region of the Philippines, is a plant from the Urticaceae family. It is commonly found in the mountain areas of Taiwan and the Philippines. This study aims to determine the Dendrocnide meyeniana root extract's anti-angiogenic activity. The methanolic extracts of Dendrocnide meyeniana were subjected to preliminary phytochemical screening. Cytotoxicity test using Brine Shrimp Lethality Assay was conducted with different plant concentrations to determine the concentration to use for the anti-angiogenic activity of the root extracts. The anti-angiogenic activity was observed by calculating blood vessel percent inhibition using chorioallantoic membrane assay or CAM assay. Phytochemical screening reveals that the plant contains varying levels of detection of bioactive compounds, including saponins, flavonoids, alkaloids, and steroids. Brine Shrimp Lethality Assay showed that the ethyl acetate and hexane extracts from Dendrocnide meyeniana were highly toxic as their LC₅₀ values were lower than 100 ppm. CAM assay results showed that hexane with 250 ppm concentration has the highest percent inhibition with 34.98% followed by hexane at 125 ppm with 34.07% inhibition. Both concentrations of ethyl acetate showed low percentage inhibition, where 250 ppm was at 19.05% and 125 ppm at 15.93%. One-way ANOVA indicates that the different treatments have significant differences (p-value< 0.0001) in the number of branches present using the CAM assay. Therefore, based on the results, Dendrocnide meyeniana root extracts have anti-angiogenic properties and these findings will help to understand the efficacy of the traditional medicine used by the local people.

Molecular Detection and Identification of Begomovirus Infecting Cucumber (Cucumis sativus) in Terengganu, Malaysia

Sakthivel Poraya Goundar — 2024-06-30

The genus Begomovirus from the family Geminiviridae is responsible for causing significant economic losses to many important horticultural crops, including cucumber (Cucumis sativus L.). Begomovirus infection during the early stages of plant growth can lead to complete yield loss. Hence, the identification of begomovirus species is important to design a precise resistant breeding strategy. This study aims to detect the presence of begomovirus in typical symptomatic cucumber leaves, identify the species of begomovirus present, and investigate the evolutionary relationships with other reported begomoviruses using phylogenetic analysis. Leaf samples from symptomatic cucumber plants were collected from the Green World Genetics (GWG) research station and a farm in Lembah Bidong, Rhu Tapai, Setiu, Terengganu. To detect the presence of begomovirus, PCR was carried out using universal primers targeting DNA-A, DNA-B, and betasatellite regions. DNA-A and betasatellite fragments were amplified, but not DNA-B. The amplified partial sequences of DNA-A were then analysed and compared with other begomovirus sequences in the GenBank database managed by the National Centre for Biotechnology Information (NCBI). The newly isolated DNA-A sequence from cucumber was 100% identical to the tomato leaf curl New Delhi virus (ToLCNDV). The phylogenetic tree was divided into two groups: group A, consisting of the newly isolated DNA-A sequence from cucumber, ToLCNDV, followed by squash leaf curl China virus (SLCCNV) and a monopartite begomovirus Ageratum yellow vein virus (AYVV); and group B, consisting of tomato yellow leaf curl Kanchanaburi virus (TYLCKaV), pepper yellow leaf curl Indonesia virus (PepYLCIV), and pepper yellow leaf curl Aceh virus (PepYLCAV). For the first time in cucumber, betasatellite is reported in association with ToLCNDV, a bipartite old-world begomovirus. This study provides a basis for the selection and breeding of begomovirus-resistant cucumber varieties in the future.

Degradation of Polypropylene Using Fungal Enzyme As A Sustainable Approach To Management Plastic Waste

Ghim Hock Ong — 2024-06-30

Polypropylene (PP) is a major environmental problem in Malaysia because it has been ranked the 28^th highest plastic polluter in the world (at 56kg per capita per year) in 2021. Landfilling is one of the most common ways of dealing with plastic because leachate may cause increased probability of cancer and neurological impairment in humans. The use of fungi in mycoremediation makes the process eco-friendly. In addition, fungi have a vast hyphal network and broader metabolic competence. The objective of this study was to investigate fungi remediation of PP via the detection of manganese peroxidase and laccase activity in Bushnell Haas Broth (BHB). PP degradation activity was measured via the activity of laccase and manganese peroxidase at a wavelength of 450nm and 610nm, respectively. Of the 17 species of fungi isolated from the Jeram landfill, 12 species of fungi showed growth in BHB with PP as the sole carbon source. Penicillium sp. 1, Aspergillus sp., Penicillium levitum, Talaromyces louisianensis, Aspergillus tamarii, Cunninghamella bertholletiae, Penicillium sp. 2 and Aspergillus niger demonstrated high and longer laccase activity, and these fungi could be considered as potential fungi. P. levitum, P. janthinellum, Penicillium sp, and T. louisianensis have high and longer MnP activity. In summary, P. levitum and T. louisianensis have a high and long duration of MnP and laccase activity in degrading PP, which can be developed and integrated into plastic waste management.

Identification and Evaluation of Locally Isolated Fungi Through Rapid Screening for Potential Mycelium-Based Biofoam Application in Malaysia

Nur Liyana Mohd Izan — 2024-06-30

Expanded polystyrene foam (EPS) contributes to environmental problems due to its inability to decompose in nature within a short period. In addition, petroleum as a core source for EPS is now depleting and new biological and environmentally friendly approaches are encouraged. As an alternative to EPS, mycelium-based biofoam (MBF) is a new foam technology formed of agricultural biomass and mycelium as a binding matrix is introduced. However, based on previous literature, the fungal strains used are mainly highlighted as one of the main factors which affect the final properties of MBF. Thus, this study aims to evaluate the most potential fungus used for MBF application using OPEFB, biomass from the palm oil industry as novel substrate using rapid screening. Twelve local fungi isolated from a local forest in Selangor, Malaysia were cultivated on Potato Dextrose Agar and OPEFB plate before being screened on agar containing four different types of dye indicators, which are methylene blue, guaiacol, Remazol Brilliant Blue R and azure B. As a result, Phanerochaete concrescens isolate LYN-UPM S1 and Perenniporia subtephropora isolate LYN-UPM S9 have shown the ability to produce ligninolytic enzymes and high content of chitin, which will be useful for the fabrication of mycelium-based biofoam.

Selecting Phosphorus-Solubilizing Strains of Purple Nonsulfur Bacteria Isolated From Pineapple Cultivated Acid Sulfate Soils

Tran Ngoc Huu — 2024-07-04

The presence of acid sulfate soils is such an obstacle for pineapple cultivation in Vietnam due to their low pH, high toxicity and poor nutrient availability, especially phosphorus (P), which is immobilized by cations in the soils. Therefore, the study occurred to select purple nonsulfur bacteria (PNSB) strains that can solubilize P under toxic and acidic conditions. There were 33 strains that can tolerate the acidic condition, and they were selected and tested for viability and P solubilization under conditions containing Al³⁺, Fe²⁺, and Mn²⁺ toxins. Four strains, including W15, W39, W42 and W48 suffered from growth inhibition by Al³⁺, Fe²⁺ and Mn²⁺ less than the other strains under both microaerobic light and aerobic dark conditions (ML and AD conditions). In addition, there were four strains (W15, W25, W42 and W48) solubilizing Al-P well (21.4-25.2 mg L^-1), two strains (W23 and W42) solubilizing Fe-P well (15.9-17.3 mg L^-1), and two strains (W17 and W42) solubilizing Ca-P well (23.0-36.4 mg L^-1) under both ML and AD conditions. Ultimately, there were five strains selected (W17, W23, W25, W42 and W48) and identified as Rhodopseudomonas palustris strain W17 and W23, Cereibacter sphaeroides strain W23, W42 and W48 based on the 16S rRNA technique. The selected strains also produced ALA, EPS and siderophores at 1.31-2.19 mg L^-1, 0.78-1.89 mg L^-1, and 16.2-55.6%, respectively. Therefore, these strains were promising in providing nutrients for pineapples in the form of biofertilizer.

Genetic Variability of Oil Palm (Elaeis guineensis Jacq.) Deli Dura Inter-Crosses Population in The MPOB Research Station Hulu Paka, Terengganu, Malaysia

Ahmad Malike Fadila — 2024-06-30

The oil palm Deli dura breeding population is the most widely used dura for breeding and seed production. Due to its narrow genetic base, with current material originating from the four initial dura palms of Bogor, the Deli dura is presumed to have low genetic diversity. A total of 35 MPOB Deli dura inter-crosses from eight paternal parents were characterized based on 22 different traits including bunch yields, bunch quality components, as well as other vegetative and physiological traits. Based on the principal component analysis of the collected data, the first five components (PC1-PC5) possessing eigenvalues surpassing 1, contributed to 97.83% of the overall variance. PC1, with the highest variation (30.88%), was characterized mainly by vegetative and physiological traits, while PC2, with about 26.02% variation, was primarily associated with bunch quality components. The principal component score plot suggests that populations of the Johor Labis paternal parents 0.278/318 and 0.278/295 are unique populations and can be used in further breeding programmes. Besides, the populations from Ulu Remis (0.280/68) and Ulu Remis × Elmina (0.281/44) paternal parents were positively associated with PC2, suggesting strong performance in bunch quality components. Cluster analysis indicated that all populations were clustered into three main groups comprised of several sub-clusters, with populations of paternal parents 0.279/48 (Banting) and 0.281/74 (Banting) were outliers in the second and third clusters, respectively. In overall, the study revealed the presence of variation among the Deli dura inter-crosses based on the traits evaluated. These results will be useful for the selection of specific populations for genetic improvements of bunch yield, bunch quality, as well as vegetative and physiological characteristics.

Controlling Soil-Borne Fungus Aspergillus niger in Groundnut By Optimizing The Function of Isolated Bacillus Bacteria

Nguyen Xuan Hieu — 2024-06-30

Collar rot is a devastating disease caused by the soil-borne pathogen Aspergillus niger that greatly affects groundnut production worldwide. The long-term persistence of the fungus in the soil can reduce the effectiveness of synthetic fungicides. Recently, significant attention has been raised to the use of the biological control method such as the application of antagonistic microorganisms, which potentially decline the number of spores and eradicated A. niger from the soil. In the present study, three Bacillus strains (Bacillus siamensis 101, B. siamensis 112 and B. velezensis 137) isolated from the rhizosphere soil of groundnut cultivation farms were found to inhibit the growth of A. niger mycelia by 53.6% to 60.8% in vitro. In pot experiments, the supplementation of this mixture of three bacterial strains (namely BAZ04) strongly reduced the collar rot symptoms of groundnut with a biocontrol efficacy of 100% compared to nil (no treatment). Field trials demonstrated the efficiency of BAZ04 in controlling collar rot disease, which increased the yield by 20.5–22.7% compared to the untreated plots. These results suggest that BAZ04 is a potential biocontrol agent for collar rot disease.

Risk for Endoparasites Among Production Stages of Female Goats With Notes on Sustainable Parasite Control For Smallholder Flocks

Bura Thlama Paul — 2024-06-30

The risk of parasitism in different groups of small ruminants depends on intrinsic, environmental, and management factors. Although there are different views regarding the sex-related risk of endoparasites in small ruminants, females are undoubtedly the most affected group in the flock. Moreover, whether the greater sex-specific risk of parasitic infection observed in female goats in field situations is associated with their production or other intrinsic factors is still under scrutiny. In this paper, cross-sectional epidemiological data collected from selected small ruminant flocks were analysed to determine the distribution, risk, and burden of endoparasites in young, nonpregnant, pregnant, and lactating female goats. There was a higher incidence of gastrointestinal parasites (88.4%, 95%CI= 83.01-92.19) than blood protozoa (54.0%, 95%CI= 46.85-60.92), with a significant difference among the groups. A higher risk of gastrointestinal parasites was observed in lactating (OR = 46.667, P = 0.001) and pregnant (OR = 9.167, P = 0.003) groups. A greater risk of blood protozoan infection was also observed in the pregnant (OR = 5.971, P = 0.0104) and lactating (OR = 3.600, P = 0.0528) groups. A significant increase in the mean faecal egg count of the lactating (2.72 ± 0.76) and pregnant (2.34 ± 0.97) groups (P < 0.05) was accompanied by a significantly lower mean PCV in the lactating group (23.48 ± 4.838) than the kids (29.44 ± 6.13), or nonpregnant (27.80 ± 5.525) groups (P < 0.05). Thus, the pregnant and lactating female goats may experience a greater exposure risk and burden of endoparasites. Therefore, female goats may be selectively targeted for implementing nutritional management, controlled grazing, and selective anthelmintic treatment during pregnancy and lactation to save cost and minimise excessive use of anthelmintic.