HAYATI Journal of Biosciences
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Characterization and Enzymatic Assay of Cellulase-Producing Probiotic Bacteria Isolated from Traditional Fermented Bamboo of Bangladesh
Full text linkFermented bamboo (also known as Ngwopo) is a traditional food item among the indigenous population living in the south-east of Bangladesh. Though fermented foods harbour a significant number of probiotics and enzyme-producing bacteria, there is a lack of research on isolating beneficial bacteria from native fermented food products in Bangladesh. Thus, considering the extensive utilisation of cellulase enzymes and probiotics, our study was conducted with the objective of isolating and characterizing native cellulase-producing bacteria from fermented bamboo and performing crude enzyme activity assay. The sample was collected from Khagrachari hill tracts district of Bangladesh. Three isolated bacteria, namely FB-1, FB-2, and FB-3, screened based on the carboxymethyl cellulose medium, were tested for biochemical and probiotic properties, and the best isolate was genetically identified by molecular characterization through 16S rRNA gene sequencing and evolutionary analysis. All three gram-positive isolates were observed to be positive for starch hydrolysis and fermentation of sucrose, suggesting their ability to breakdown carbohydrates while testing negative for gelatinase, indole, and H2S test. The isolates demonstrated moderate bile salt and pH tolerance and low to medium autoaggregation. FB-2 had 75% susceptibility to the tested antibiotics, while other isolates were susceptible to 50% antibiotics. All of the bacteria were non-pathogenic, showing γ-haemolysis. FB-2 strain showed an enzyme production capacity of 1.56 units/ml. The best performing isolate was identified as Bacillus cereus. Therefore, the study discovered a native bacteria potentially suitable as probiotic and as a source of cellulase for industrial processes, highlighting the enriched indigenous cuisines of Bangladesh
Exploration of Lipid Profile and Wax Ester Content from Local Strain Euglena sp. IDN33 Cultivated in Mixotrophic Condition Combined with Molasses Supplementation
Full text linkThe transition from fossil fuels to sustainable renewable energy is currently growing rapidly. Euglena sp. can be the source of biofuel. Molasses, in addition to Euglena sp., can increase the levels of paramylon and wax esters produced. Therefore, this study aims to analyze cell growth, lipid profile, and wax ester content of Euglena sp. cultivated under mixotroph conditions with molasses. Euglena was cultivated under mixotrophic conditions with the addition of various concentrations of molasses (0, 10, 15, and 20 g/L) until the final log phase. Then, measurements were made on cell density, levels of lipids, proteins, and paramylon to measure the growth and metabolites profile of Euglena sp., along with lipid profile and wax ester analysis using GC-MS. The results of the study showed that 10 g/L molasses increased Euglena growth (SGR 0.1736± 0.0213 mg/ml) and carbohydrate (0.426 mg/ml) but reduced lipid accumulation (2.29 mg/ml). This treatment provides the best result before control. Molasses does not increase carbohydrates and protein but increases paramylon accumulation. The GC-MS assay detected more lipid profiles and concentrations of each lipid type in non-molasses-treated Euglena sp. compared to molasses-treated. Euglena was cultivated on molasses, which was dominated by palmitic acid and myristyl myristate with the absence of PUFA-type fatty acids in the cells, while the untreated ones were dominated by stearic acid and myristyl myristate. Thus, 10 g/L supplementation of molasses is the most effective treatment compared to others
The Trehalose-6-Phosphate Synthase and Trehalose-6-Phosphate Phosphatase in Cocoa (Theobroma cacao L.): Genome-Wide Identification and Expression Analysis
Full text linkCocoa (Theobroma cacao L.), a vital industrial crop renowned for its economic and nutritional significance, faces increasing challenges due to climate change-induced stresses. To enhance the understanding of cocoa\u27s adaptive mechanisms, a comprehensive analysis was conducted on the trehalose-6-phosphate phosphatase (TPP) and trehalose-6-phosphate synthase (TPS) gene families, which play crucial roles in plant stress responses and development. Five TcTPP and eight TcTPS genes were identified using the latest cocoa genome assembly, distributed unevenly across nine of the ten chromosomes. Detailed physicochemical characterization revealed significant variability in amino acid length, molecular weight, isoelectric point, and hydrophilicity among these proteins, suggesting functional diversity. Phylogenetic analyses, performed using the maximum likelihood method, classified the TcTPP family into three distinct clades and the TcTPS family into two main groups. Gene structure examination uncovered variations in exon-intron organization, with TcTPP genes containing nine to twelve exons and TcTPS genes ranging from three to eighteen exons, indicating structural diversity within these families. Based on publicly available datasets, expression profiling demonstrated differential expression patterns of TcTPP and TcTPS genes during embryo development and under biotic stress conditions, such as pathogen infection by Phytophthora megakarya. Certain genes exhibited significant upregulation or downregulation in response to stress, implicating them in cocoa\u27s defense mechanisms. Taken together, this study provides valuable insights into the TPP and TPS gene families in cocoa. It lays a foundation for developing strategies to enhance stress tolerance and sustainability in cocoa cultivation amidst changing climatic conditions
Screening and Molecular Identification of Endophytic Bacteria from Calamus caesius Blume with Potential as Antioxidant and α-Glucosidase Inhibitors
Full text linkCalamus caesius, traditionally used in Asian medicine for managing diabetes mellitus, is a promising source of endophytic bacteria due to its bioactive potential. This study aims to identify the potential of endophytic bacteria isolated from “Pakkat” (Calamus caesius Blume), a traditional food of North Sumatra, as a source of α-glucosidase inhibitors. These inhibitors, such as acarbose, are used to manage type 2 diabetes mellitus but have significant gastrointestinal side effects. This study focuses on isolating and characterizing endophytic bacteria that can produce natural inhibitors, potentially safer alternatives. Twenty endophytic bacterial isolates were successfully isolated, of which five were excluded due to hemolytic activity. Antioxidant activity was determined using the DPPH method, and isolates with the highest activity proceeded to α-glucosidase inhibitor testing. Molecular identification of potential isolates was performed through 16S rRNA gene amplification, and genomic DNA sequencing utilized the Oxford Nanopore Technologies platform. Secondary metabolite analysis was performed using antiSMASH, and gene cluster comparison with CAGECAT. Of 15 isolates, LTUP1.3 (Bacillus cereus) exhibited 86.6% α-glucosidase inhibitor activity, close to the positive control (88%). Although in Bacillus cereus LTUP1.3 no secondary metabolites were found that act as α-glucosidase inhibitors, three enzymes have similarities with enzymes found in Actinoplanes that play a role in acarbose biosynthesis namely dTDP-glucose 4,6-dehydratase RffG, glucose-1-phosphate thymidylyltransferase rmIA and glucose-1-phosphate adenylyltransferase glgC. These results suggest that Bacillus cereus strain LTUP1.3 has a potential biosynthetic pathway similar to the acarbose biosynthetic pathway in Actinoplanes. However, further confirmation is required to establish its overall acarbose production capability
Drought-Induced Morphophysiological and Metabolite Changes in Oil Palm Seedlings from Jambi, Indonesia
Full text linkOil palm requires sufficient water for optimal growth. However, oil palm plantations in Indonesia face the challenge of global climate change, which causes increased temperatures and unpredictable rainy seasons. The research aimed to analyze oil palm accessions from Jambi that are tolerant to drought based on plant morphophysiological characteristics and metabolite profile. The experiment was conducted in the greenhouse using a factorial randomized block design. Five oil palm accessions were evaluated in this study, i.e., Muara Jambi (MJ), Tebo (TB), Tanjung Jabung Barat (TJB), Merangin (MR), and Simalungun (SM). Seedlings were planted in pots containing 20 kg of soil and acclimatized for 14 days before treatment. Drought treatments were carried out for two, four, and six weeks. Observations were made on growth, physiological, and metabolite changes at the end of each drought period. The study showed that the SM accession exhibited a higher photosynthetic rate and fresh and dry biomass than the other accessions. However, the content of proline and MDA increased significantly in oil palm throughout the drought periods. Metabolomic profiling revealed a significant increase in 13 metabolites under drought stress, i.e., citramalate, L-tyrosine, ferulate, and 3-4-dihydroxybenzoate were the four most prevalent metabolites in the leaf. The root produced more L-proline, aspartate, aconitate, kaempferol-3-o-pentoside, oxoadipate, citrate, L-phenylalanine, maltose, phenylacetic acid, and citramalate. These metabolites have roles in plant metabolic pathways such as valine, leucine, and isoleucine biosynthesis, phenylpropanoid biosynthesis, arginine and proline metabolism, citrate cycle (TCA cycle), lysine biosynthesis, phenylalanine metabolism, and starch and sucrose metabolism
Comprehensive Characterization of Phospholipase C and D Families in Cocoa (Theobroma cacao L.): Identification, Phylogenetics, Gene Structure, and Transcriptomic Insights
Full text linkThis study provides a detailed investigation of the phospholipase C (PLC) and phospholipase D (PLD) gene families in cocoa (Theobroma cacao), focusing on their identification, characterization, and expression patterns. A total of 10 PLC and 12 PLD genes was identified and systematically annotated based on their sequence homology, conserved domains, and functional classification, adhering to established nomenclature. Analysis of physicochemical properties revealed diversity in molecular weights, isoelectric points, and stability parameters, reflecting their structural and functional variability. Phylogenetic analysis classified the genes into distinct subfamilies and highlighted their evolutionary relationships with homologs in Arabidopsis thaliana and rice (Oryza sativa). Gene structure analysis demonstrated significant variation in exon-intron organization, indicating functional specialization and regulatory complexity within these gene families. Expression profiling during cocoa embryo development showed that certain genes, such as TcNPC2, TcPI-PLC5, and TcPLDα1, were highly expressed, while others exhibited stage-specific activity. In response to Phytophthora megakarya infection, several PLC and PLD genes displayed significant changes in expression across different time points and genotypes, including the upregulation of TcPI-PLC2, TcPLDα5, and TcPLDζ2, suggesting their roles in cocoa\u27s stress responses and defense mechanisms. These findings offer new insights into the biological roles of PLC and PLD gene families in cocoa, particularly in growth, development, and stress adaptation, providing a solid foundation for further functional research and potential applications in cocoa improvement programs
The Gene Analysis and Probiotic Potential Characterization of Pseudomonas alcaligenes SG03
Full text linkAHPND causes up to 80% of shrimp mortality, severely impacting Indonesia\u27s aquaculture. Rising antibiotic resistance necessitates sustainable alternatives. This study investigates the probiotic potential of Pseudomonas alcaligenes SG03, a bacterium with unique genomic traits, to address antibiotic resistance and promote animal health. Genome analysis revealed a 6.17 Mb genome with 4,446 protein clusters, including genes for carbohydrate metabolism (26.63%), amino acid metabolism (19.93%), and energy metabolism (10.88%). Key probiotic-related genes, such as acid resistance (rpoS, actP), salinity tolerance (gshA, cysK), antibiotic resistance (vanB, gyrA), and metal resistance (copZ, zwf), were identified. In vitro, assays evaluated probiotic properties under conditions mimicking the shrimp gastrointestinal tract. Osmo-tolerance tests showed optimal growth at 10% glucose, with a significant drop in viability at higher concentrations. Autoaggregation increased progressively, reaching 0.78 at 18 hours and 0.13 at 48 hours. Optimal growth occurred at 30°C, with a 2.5-fold increase in optical density compared to 25°C. NaCl tolerance peaked at 2% (20.33×1012 CFU at 48 hours), while bile salt tolerance was highest at 0.1% (27.00×1012 CFU at 48 hours). pH tolerance was optimal at pH 5 (20.00×1012 CFU at 48 hours). Antioxidant (53.00%) and anti-inflammatory (60.33%) activities peaked after 48 hours. Phenotypic antibiotic resistance was observed against Tetracycline, Amoxicillin, Cefixime, Streptomycin, and Chloramphenicol, with proteolytic activity but no lipolytic or hemolytic properties. These findings highlight P. alcaligenes SG03’s potential as a probiotic in aquaculture to support shrimp health and reduce antibiotic dependency. Future studies should focus on in vivo validation and formulation development
Immune Response to Capsular Polysaccharide of Streptococcus pneumoniae in Rabbits Immunized with Pneumococcal Conjugate Vaccine
Full text linkTo evaluate the immune response of a rabbit model to polysaccharide capsules of Streptococcus pneumoniae after primary and booster immunization with pneumococcal conjugate vaccine. Rabbits were immunized with 0.25 ml of the 13-valent pneumococcal conjugate vaccine (PCV13) on Day 0, 7, and 14. Blood of rabbits was collected to measure the specific antibodies against the native polysaccharide capsules of S. pneumoniae serotypes 6B (Anti-Pn6BPS antibodies) and 19F (Anti-19FPS antibodies) using enzyme-linked immunosorbent assay (ELISA). The specific anti-Pn6BPS and anti-Pn19FPS antibodies in the immunized group increased significantly compared to the control rabbit after immunization (P<0.05). The highest titer of anti-Pn6BPS and anti-Pn19FPS antibodies were obtained on Day 21 and Day 28, respectively. The third immunization on Day 21 is a fairly high increase in specific antibodies against polysaccharide type 6B and type 19F. The highest antibody titers against polysaccharide capsules of S. pneumoniae were obtained after booster immunization
Relative abundance, activity pattern and habitat suitability of Great Argus (Argusianus argus grayi) in Sungai Wain Protected Forest, Balikpapan, East Kalimantan
Full text linkGreat Argus (Argusianus argus) is a globally threatened species that is protected by law in Indonesia. This species is a habitat specialist that tends to be confined to primary forests and is susceptible to human disturbance and environmental change. Sungai Wain Protected Forest (SWPF) is an isolated lowland rainforest in Kalimantan. The rising deforestation rate has threatened species sustainability in recent decades, including the Great Argus in SWPF. This study aimed to systematically analyze camera trap data on the Great Argus\u27s abundance, activity patterns, and habitat suitability prediction in SWPF. We used camera trap data from 2018-2022 at the SWPF. Great Argus\u27s lowest relative abundance index was in 2020, and the highest was in 2022. Overall, the relative abundance index of the Great Argus was approximately 3.125 independent events per 100-day trap night. The activity pattern of the Great Argus started at dawn and then constantly decreased until dusk. The habitat suitability prediction for the Great Argus was mostly in primary forests. Distance to road and building were the variables that contributed the most to the results of habitat suitability modeling. Habitat loss and suitability are the determining factors for the sustainability of significant argus populations
Congenital Malformations in Chicken Embryos After Oxybenzone Exposure
Full text linkTopical use of oxybenzone, commonly found in sunscreens, can be absorbed by the skin, and long-term use may cause endocrine disruption, cancer, and teratogenic effects. However, its potential teratogenic effects on embryonic development have not been well-studied. This study aims to determine the impact of oxybenzone exposure on the early stage of embryonic development. Chicken embryos aged 72 hours (20 Hamburger-Hamilton/HH stage) were exposed to a pure oxybenzone for 24 hours at varying concentrations (0, 1, 5, 10, and 20 ppm), each group consisting of 3 embryos. Embryo preparations were made using the wholemount method. Morphological abnormalities were observed with a stereo microscope, and descriptively morphometric measurements were analyzed using ImageJ software. Statistical analysis used One-way ANOVA and Tukey’s test for normally distributed data, while Kruskal-Wallis H and Mann-Whitney U test for non-normally distributed data. This study found that oxybenzone significantly enlarged the embryo, telencephalon, and eye. Several abnormalities were observed in the embryos exposed to oxybenzone, including incomplete closure of the anterior neuropore, concavity in the anterior and lateral of the mesencephalon, and depressions in the tail bud. This study concludes that oxybenzone acts as a teratogen, causing abnormalities in embryonic development, particularly in the central nervous system