HAYATI Journal of Biosciences
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Metagenomic Analysis of Bacterial Communities in the Musi River Estuary, South Sumatra, Indonesia
Get PDFUnderstanding the diversity, function, and ecological dynamics of bacterial communities depends on unique transitional environments, estuaries. Knowledge on bacterial taxa in the Musi River estuary, South Sumatra still eludes, however. This effort aims to identify bacterial species in estuary waters using metagenomic analysis based on 16S rRNA gene. Surface water samples from three study locations were analysed by DNA isolation, nanodrop spectrophotometer qualitative assessment, 16S rRNA gene amplification, electrophoresis, and Illumina NovaSeq sequencing. Results showed that Proteobacteria predominated at all sites, followed by Campilobacterota, Cyanobacteria, and Bacteroidota. At the class level, Gammaproteobacteria was most common, followed by Alphaproteobacteria and Campylobacteria. Dominant bacterial orders were Campylobacterales, Rhodobacterales, and Pseudomonadales while the most common families were Arcobacteraceae, Rhodobacteraceae, and Pseudomonadaceae. The most plentiful genera were Rheinheimera, Pseudomonas, and Pseudarcobacter. Variations in bacterial spread among stations suggest environmental factors including salinity, nutrient availability, and human activities influencing microbial community composition. Ternary plots, heat maps, and krona diagrams were employed to disclose distinct patterns of bacterial community dispersion in the estuary. This paper underscores the importance of metagenomic research in illuminating microbial diversity in estuarine environments and its impact on ecological dynamics and water quality
Indonesian Herb Extracts Inhibit the Replication of Bovine Respiratory Syncytial Virus: In Vitro Study
Get PDFBovine respiratory syncytial virus (BRSV) is highly prevalent in cattle. It is a major viral cause of bovine respiratory disease complex, which is associated with morbidity, mortality and substantial economic impact. Currently available treatments are only symptomatic, but no specific treatments are available for BRSV infection. This study aimed to identify new antiviral agents against BRSV, which could be used to control bovine respiratory disease complex in cattle with Indonesian herb extracts. Ethanol extracts prepared from Indonesian herbs including Andrographis paniculata, Phyllanthus niruri, Curcuma aeruginosa, and Curcuma xanthorrhiza were evaluated for anti-BRSV activity in A549 cells. The cytotoxicity of the herb extracts was evaluated using a CCK-8 cell viability assay. Antiviral activities of the herb extracts were examined using cell activity and cytopathic assays. The effect on virus production was evaluated by qRT-PCR and plaque-formation assays. Extracts of Curcuma xanthorrhiza (125 μg/ml), Andrographis paniculata (250 μg/ml), and Phyllanthus niruri (62.5 μg/ml) inhibited BRSV activity in A549 in pre-, simultaneously-, and post-infection treatment assays, respectively, as measured by the selective index. Reduction of BRSV activities by the herb extracts correlated with inhibition of viral gene expression and inhibition of plaque formation in a concentration- and time-dependent manner. Our findings suggest that these herb extracts have sufficient potency to be used not only as a therapeutic agent but also as a preventive agent to limit BRSV infection
The Invasive Plant Maeopsis eminii: A Key Dietary Resource for Primates in the Tropical Rainforests of West Java, Indonesia
Get PDFAnimal-plant interactions are crucial for understanding the ecological dynamics and predicting ecosystem changes. This study explores the role of the invasive plant Maesopsis eminii in the Bodogol Nature Reserve, Gunung Gede National Park, West Java, Indonesia, focusing on its interaction with diurnal vertebrate species. Primates, including Javan gibbon and Javan langur, were identified as primary consumers of Maesopsis eminii, with Javan gibbon showing the highest consumption rates. Dietary analysis revealed that both primate species utilized M. eminii fruits and leaves, whereas Javan gibbons relied more on Maesopsis eminii than Javan langurs. Other vertebrates, such as squirrels and birds, are occasional visitors with minimal fruit consumption. This study highlights that Maesopsis eminii is a primary resource of Javan gibbon, Javan langur, and Javan surili. This nearly year-round fruiting plant is crucial for supporting primate populations, especially given its contribution to their diet. Considering the mutualistic interaction between Maesopsis eminii and its visitors, the Javan gibbon was the only species that probably dispersed seeds through endozoochory, making this primate species a crucial part of the dispersal of Maesopsis eminii in Gunung Gede National Park. Because invasive plants can threaten native flora and fauna, future studies must emphasize balancing the beneficial and detrimental effects
Truncation on N-Terminal Hydrophobic Domain of L1 Major Capsid Protein of Human Papillomavirus Type 52 Enhances Its Expression in Hansenula polymorpha
Get PDFHuman papillomavirus (HPV) infection is the main cause of cervical cancer. The administration of the HPV prophylactic vaccine, which is commonly produced based on HPV L1 major capsid protein, significantly reduces the incidence of cervical cancer. However, the coverage of the HPV vaccination program is often hindered due to its relatively high cost. This study aimed to evaluate the impact of N-terminal hydrophobic domain truncation on the expression of L1 major capsid protein of HPV type 52 in Hansenula polymorpha. The truncation enhanced the yield of L1 protein expression compared with the full length, which was confirmed by Western blot and ELISA. Furthermore, the truncated L1 protein formed virus-like particles (VLPs), which were confirmed by transmission electron microscopy (TEM). Bioinformatics analysis showed that the truncated L1 protein was more soluble compared with the full length, possibly increasing the protein expression. These findings could pave the way for the development of a more cost-effective HPV type 52 L1 protein production in H. polymorpha to be used as a VLP-based prophylactic vaccine
Development of a Fluorescence Immunoassay Based on Curcumin Carbon Dots-labeled IgY Antibodies for SARS-CoV-2 Detection
No full textCarbon dots (CDs) are widely utilized in biomedical applications as fluorescent labels for imaging and diagnostics due to their excellent biocompatibility and superior optical properties. These advantages often make CDs a substitute for organic fluorescent dyes, which suffer from low emission intensity and poor photostability when interacting with biomolecules. Moreover, carbon-based materials are eco-friendly and can be synthesized from natural sources, such as curcumin, a chromophore compound abundantly available in Indonesia. Therefore, this study conducted a preliminary investigation on curcumin CDs-labeled IgY antibodies (IgY-cur CDs) for fluorescence immunoassay of SARS-CoV-2 in rapid test applications. The synthesis of the fluorescent label involved a carbodiimide coupling reaction using EDC/NHS agents to conjugate IgY antibodies with curcumin CDs. The IgY-cur CDs conjugate was confirmed to detect antigens through FRET immunosensor mechanisms, showing a significant increase in fluorescence intensity with increasing antigen concentrations (p < 0.05), with a minimum sample concentration of 10 ng. Furthermore, the IgY-CDs cur conjugate was applied as a reporter in a fluorescence-based LFIA using a sandwich assay format. The test strip successfully detected synthetic multiepitope SARS-CoV-2 antigens with an estimated detection limit of 54.28 µg and nasopharyngeal samples from confirmed COVID-19 patients within 35 minutes of operation. The test strip was evaluated to be stable under cold storage at 4°C for up to 3 weeks. In conclusion, curcumin CDs-labeled IgY antibodies demonstrate promising potential for further development as fluorescent labels in rapid diagnostic applications targeting SARS-CoV-2
Endophytic Entomopathogenic Fungi Negatively Impact on Growth and Development of Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae)
No full textAn alternative method for controlling Spodoptera frugiperda applies entomopathogenic fungi (EPF). The present research aimed to determine the impact of the endophytic EPF on the growth and development of S. frugiperda. Twenty molecularly identified isolates of the endophytic EPF were used in seed treatments with the fungal suspension 1 × 1010 conidia.mL-1. The endophytic EPF that colonized corn leaves eaten by the neonate S. frugiperda larvae was able to significantly decrease the body weight of the larvae and pupae. The endophytic EPF could prolong the developmental time of all instar larvae and pupae. The endophytic EPF could raise larval and pupal mortality and reduce the ability of adults to emerge from pupae, and decrease egg laying by females. The lowest percentage of adult emergence was observed in the adults from larvae consuming corn leaves colonized with Beauveria bassiana JaGiP and JgSPK isolates, which were 34.67% and 24%, respectively. Consequently, a reduced adult emergence could lead to a high cumulative death rate of 76% from larvae to adults. Finally, the endophytic EPFs negatively affect S. frugiperda growth and development. B. bassiana JaGiP and JgSPK isolates are the most pathogenic fungi in inhibiting the growth and development of S. frugiperda. They are promising candidates for controlling S. frugiperda in the corn field.
Abundance and Season Variability of Rove Beetles (Coleoptera: Staphylinidae) in the Mangrove Ecosystem of Akalapuzha Coastal Region, Kerala, South India
No full textThe species composition and diversity of rove beetles (Staphylinidae) in the mangrove coastal region remain poorly explored, particularly in South India. This study aims to understand the rove beetle diversity in the Akalapuzha mangrove coastal ecosystem with respective seasonal changes (pre-summer, summer, and monsoon) as well as the efficacy of different collection methods (pitfall trap, light trap, Berlese funnel, and flotation method). From the study, the collected specimens came under five subfamilies: Oxytelinae, Aleocharinae, Staphylininae, Paederinae, and Tachyporinae. Among this, high taxa abundance was observed in Oxytelinae, followed by Aleocharinae, Staphylininae, Paederinae, and Tachyporinae. Among the documented subfamilies, three subfamilies, namely Oxytelinae, Staphylininae, and Aleocharinae were recorded in all three seasons. In the case of diversity analysis, the highest diversity was observed in the summer season (1-D = 0.277, & H = 0.630), followed by pre-summer (1-D = 0.306, & H = 0.678) and monsoon (1-D = 0.533, & H = 1.069). Among all the different insect collection methods, the flotation method (p<0.05) is the most suitable for collecting rove beetles, regardless of seasonal variations, and is followed by light traps, pitfall traps, and Berlese funnel traps. The study revealed the inevitability of protecting the mangrove ecosystem, as it is identified as an ideal habitat for the economically, ecologically, and medically important Coleopteran family, Staphylinidae
Entomological Assessment of Mosquito Diversity and Density in Negara District, Bali: Implications for Vector-Borne Disease Surveillance
No full textIndonesia has one of the highest prevalences of malaria in Asia, with annual case numbers on the rise. Despite being a major international tourist destination, the Negara Sub-district in Jembrana Regency, Bali, continues to experience notable incidences of both diseases. Entomological surveys are crucial for evaluating mosquito diversity and density, which supports effective vector control strategies. This study, conducted in five villages from September to October 2024, employed entomological survey methods to calculate the House Index (HI), Container Index (CI), Breteau Index (BI), and Density Figure (DF). Mosquito species diversity was assessed using the Shannon-Wiener diversity index and evenness (E) and dominance (C) metrics. Results indicated that HI, CI, and BI values were within the low transmission risk category, with a DF of 4 and a larva-free rate (ABJ) of ≤95%. Six mosquito species were identified: Aedes aegypti, Aedes albopictus, Anopheles dirus, Armigeres subalbatus, Culex quinquefasciatus, and Mansonia uniformis. The species diversity was moderate (1 < H\u27 ≤ 3), even species distribution (E > 0.6), and low dominance (C ≤ 0.5). These findings suggest a relatively low risk of vector-borne disease transmission. However, given the presence of multiple disease-vector species and the region\u27s dynamic human activity, continuous entomological surveillance remains essential to anticipate population surges and strengthen disease prevention efforts
Enhanced Lipase Production in Pichia pastoris via Multiple Copies of Bacterial Lipase Genes and Co-expression of the HAC1 Gene
No full textThe hac1 gene, a key regulator of the untranslated protein response (UPR), was co-expressed in Pichia pastoris GS115 to enhance the production of a lipase from Geobacillus stearothermophilus. Multicopy lipase constructs (1X and 4X) were transformed with pPICZAwbe_hac1, generating GS115/T1.2RQ(1X)_hac1 and GS115/T1.2RQ(4X)_hac1 strains. The GS115/T1.2RQ(1X)_hac1 strain showed an 186% lipase activity after 120 hours versus the control (100%), while the GS115/T1.2RQ(4X)_hac1 strain showed a faster initial increase (38% at 48 hours) and 28% at 120 hours, which was beneficial for efficient enzyme production. Overexpression of the hac1 gene enhances lipase production because it activates UPR genes when the endoplasmic reticulum is stressed due to a large number of recombinant proteins and forms proteins that are not appropriately folded. SDS-PAGE and tributyrin plate assays confirmed extracellular lipase expression (~43 kDa). These results demonstrate that hac1 co-expression significantly (p = 0.01) enhances lipase production in Pichia pastoris, especially in lower-copy constructs. This is the first report of co-expressing hac1 with Geobacillus stearothermophilus lipase genes in yeast. The findings are expected to contribute to developing more efficient microbial cell factories for producing industrial enzymes
Molecular Characterization of Infectious Bursal Diseases Virus VP2 Gene Fragments Obtained from Commercial Broiler Farms in Central Java and The Yogyakarta Special Region Province
Get PDFInfectious Bursal Disease (IBD) is an infectious and immunosuppressive disease primarily affecting young chickens. Despite stringent biosecurity and vaccination for control measures, the effective management of IBD remains challenging. The disparity in observed clinical symptoms in the field infections further complicates matters for breeders. The study aims to perform molecular characterization of VP2 gene fragments to identify the latest genotype of field IBD viruses. Twenty-two samples of bursa of Fabricius were collected from broilers suspected of IBD in commercial farms located in Central Java and The Yogyakarta Special Region from 2021 to 2022. Viral RNA was extracted from these samples, and after amplification, a 743 bp PCR product was obtained and subjected to sequencing. The obtained sequences were analyzed in Mega X for multiple alignments, amino acid prediction, homology, and phylogenetic tree construction. Lesion, i.e., Bursa of Fabricius enlargement, oedema, swelling of plica bursa, gelatinous mass, hemorrhage, atrophy, and thigh muscles petechiae to hemorrhage, were considered indicative of IBD. Out of 22 samples tested by RT-PCR, 19 were positive, and 13 samples were selected for sequencing. All sequenced samples belonged to Genogroup A3, specifically the very virulent IBD (vvIBD) strain