HAYATI Journal of Biosciences
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Successful Primer Picking and Pooling for the Design of Multiplex PCR Primers Specific to Pork, Beef, Chicken, and Rat DNA
Get PDFDNA markers and Multiplex-PCR have emerged as methods for species detection in processed meat products. The primary objective of this study is to design multiplex primer sequences for pork, rat, beef, and chicken, generating distinguishable amplicons through agarose gel electrophoresis for halal detection in processed meat products. Primer design involved utilizing mitochondrial genomic data and the NCBI-Primer BLAST site to obtain specific pork and beef primer sequences. In silico simulations, including single and multiplex-PCR, were conducted using Primer Pooler. In vitro validation encompassed Single-PCR and Multiplex-PCR annealing temperature optimization, using samples of chicken, beef, pork, and rat as well as processed meat products like meatballs, sausages, and nuggets. In vitro validation demonstrated that the halal marker gene\u27s multiplex primer efficiently amplified the target sequence, specifically at the optimal annealing temperature of 58°C. Amplicons from beef (1,217 bp), pork (860 bp), rat (622 bp), and chicken (272 bp) primers could be distinguished on a 1.5% agarose gel. The study\u27s results can aid in cost-effective and rapid halal testing and authentication of processed meat products, offering advantages over PCR with a single primer
Endoparasites of Wild Javan Gibbon (Hylobates moloch) At Gunung Halimun Salak National Park, Indonesia
Get PDFInfections of endoparasites in primates in natural habitats are highly prevalent and can cause disease, reduce health quality, and disrupt their life. This study investigated endoparasites prevalence value in the endangered Javan gibbon (Hylobates moloch) in Citalahab Forest, Gunung Halimun Salak National Park, Indonesia, from June to August 2022 by collected fecal samples (N = 10) and analyzed it using floatation methods. As a result, we found five genera of nematodes Trichuris trichiura (10% egg worm prevalence, Oesophagostomum spp. (50%), Trichostrongylus spp. (60%), Ancylostoma spp. (80%), and Strongyloides spp. (100%). The prevalence value of the worms in the larvae stage of Trichostrongylus spp. 20% and Strongyloides spp. 70%. Nematode infection status successively is Strongyloides spp., which is, frequently; Trichostrongylus spp. and Ancylostoma spp., which is, often; Oesophagostomum spp. and Trichuris trichiura which is, occasionally. Four species of nematode were found in both ages, and only Trichuris trichiura was found in one adolescent individual. The threat posed by this parasite deserves attention; further research is needed to fill the gap in our knowledge of their pathogenicity and transmission in Javan gibbon
Isolation and Identification of Vibrio parahaemolyticus Bacteria in Bottlenose Dolphins (Tursiop truncates) in Kendal Conservation Pond, Central Java
Get PDFAquatic mammals in Indonesia are officially protected. However, there is a lack of research on these mammals, particularly in relation to potential disease-causing bacteria. A study was conducted in a conservation pond to address this gap, where swab samples were taken from the blowholes of aquatic mammals. The focus was on identifying bacteria that could potentially cause infectious diseases in these animals. The results revealed V. parahaemolyticus bacterial isolates, which showed a 98% similarity to the registered V. parahaemolyticus in NCBI. These bacterial isolates exhibited hemolysin properties and demonstrated resistance to trimethoprim, streptomycin, cephalothin, and penicillin antibiotics
Antioxidant, Antidiabetic, and Antibacterial Activities of Terminalia bellerica Seed Extracts in Various Solvent Polarities
Get PDFTerminalia bellerica is well-known for producing edible fruits with pharmacotherapeutic properties. Traditional healers use these species to treat and control diabetes mellitus, its side effects, and other illnesses. Involved in this disease\u27s pathophysiological process, extensive research has been conducted to validate and comprehend these bioactive claims scientifically. This research aims to ascertain the bioactive metabolite contents of different solvent polarities, including ethyl acetate, hexane, distilled water, and methanol. The phenol concentration was determined using the Follin-Ciocalteu procedure to be between 23.45 and 160.41 mg GAE/g. The aluminum chloride colorimetric technique measured flavonoid concentration from 88.52 to 7.12 mg QE/g. The quantitative values of tannic acid, which spanned from 0.78 to 5.32 mg TAE/g, were determined by spectrophotometry UV-VIS. The extracts\u27 capacity to reduce free radical damage ABTS (2, 2\u27 azinobis (3-ethylbenzene-thiazoline-6-sulfonic-acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) was examined. The extract ethyl acetate exhibited the most significant level of antioxidant activity, with IC50 values for DPPH and ABTS of 28.17 and 22.22 g/ml, respectively. Staphylococcus aureus (23 mm) and were tested for antibacterial and antifungal activity in a methanol extract (21 mm). In vitro, antidiabetic activities were assessed using α-glucoside and α-amylase inhibition. The ethyl acetate extract has α-glucoside inhibition IC50 of 23.04 g/ml and α-amylase inhibition IC50 of 25.35 g/ml. T. bellerica seed includes secondary metabolites that show promise as lead chemicals in creating potent medications
Cloning, Expression, and Bioinformatics Modeling of Human Papillomavirus Type 52 L1/L2 Chimeric Protein in Escherichia coli BL21 (DE3)
Get PDFHuman papillomavirus (HPV) L1 major capsid protein generates a highly immunogenic virus like particles (VLPs), which have been used as the main component of its prophylactic vaccine. However, the neutralizing antibodies against L1 VLPs are mostly type specific and may not be effective to prevent infection from different strains of HPV. On the other hand, HPV L2 minor capsid protein has low antigenic variation, thus can induce cross-neutralization. This study aims to obtain HPV 52 L1/L2 chimeric protein, which is designed based on HPV type 52 as one of the most circulated high-risk types in Indonesia, to develop a broad-spectrum HPV vaccine. Substitution of HPV 52 H4 helix L1 region with an HPV 52 L2 epitope was carried out using overlap extension PCR. HPV 52 L1/L2 chimeric gene was constructed into pET-SUMO expression vector and expressed in Escherichia coli BL21 (DE3). Bioinformatics modeling suggested that L2 epitope was located inside of the loop region in monomer form, and on the contrary, it was located outside of the pentamer surface. Furthermore, B cell and T cell epitopes predictions were conducted using Immune Epitope Database (IEDB) analysis. The B cell epitopes prediction revealed eleven potential epitopes, whereas the T cell epitopes prediction showed seven potential epitopes for each MHC class I and MHC class II. This study showed that HPV 52 L1/L2 chimeric protein has the potential to induce cross-neutralizing antibodies and can be developed as a promising candidate for a new HPV vaccine
The Effect of Enhancing Ulvan\u27s Antioxidant Properties in Supplemented Diets on Accelerating The Phenoloxidase Immune Response in White Shrimp
Get PDFThis study investigated the total phenol content (TPC) and DPPH inhibition of ulvan from Ulva lactuca, along with its supplementation effects on the specific growth rate (SGR) and phenoloxidase (PO) activity of Litopenaeus vannamei. Ulva lactuca samples were processed to obtain four ulvan extracts using different methods such as P-HWE (Polysaccharide-Hot Water Extract), O-HWE (Oligosaccharide-Hot Water Extract), P-A-HWE (Polysaccharide-Acid-Hot Water Extract), and O-A-HWE (Oligosaccharide-Acid-Hot Water Extract). The U. lactuca powder was treated with 80% ethanol overnight, then centrifuged and dried. Depigmented U. lactuca was extracted with water (65°C, 2 hours), and polysaccharides were precipitated with 99% ethanol, yielding P-HWE. P-HWE was heated at 145°C for 4.5 hours to yield O-HWE. Another extraction after depigmentation, using water containing H2O2 and ascorbic acid (65°C, 2 hours), precipitated P-A-HWE, which was dried and heated to yield O-A-HWE. FTIR analysis provided insights into the chemical composition and structural characteristics of ulvan extracts. TPC and DPPH inhibition were measured spectrophotometrically, and PO activity using a colorimetric assay with L-DOPA. Ulvan supplementation in shrimp feed was evaluated for growth and immune response. Results showed significant differences in TPC, DPPH inhibition, and PO activity among extracts, with O-A-HWE having the highest phenol content and DPPH inhibition. Ulvan supplementation significantly influenced the SGR of L. vannamei, with the highest SGR in the O-A-HWE treatment. This study suggests that ulvan extracts, especially O-A-HWE, could be effective natural immunostimulants for enhancing the health and growth of L. vannamei, warranting further research to optimize extraction methods and understand the underlying mechanisms
Assessment of Genetic Diversity Using Morphological and Molecular Characteristics of Indonesian Zoysiagrass Genotypes
Get PDFZoysiagrass is warm-season turfgrass thriving in tropical regions. Despite the adaptive nature, the existence of Indonesian zoysiagrass as well as morphological and genetic characteristics are not available. Therefore, this study aimed to explore Indonesian zoysiagrass from western and eastern parts of the country, as well as its morphological and molecular characteristics. Morphological characteristics was conducted to measure vegetative and reproductive characters while genotyping was performed using 15 simple sequence repeat markers. Morphological characteristics cluster three major groups, namely Group 1 corresponded to short, shorter, and fine leaves. Group 2 corresponded to tall, longer, and fine leaves, fewer seeds, and short spikelet, while Group 3 corresponded to tall, long, and wider leaves, more seeds, and longer spikelet. The results showed that the expected heterozygosity (He = 0.256) was lower than homozygosity (Ho = 0.341). The high level of discriminating capacity, polymorphism, and informativeness of SSR marker was observed (Effective Multiplex Ratio = 4.20, Marker Index = 2.394, and Resolving Power = 1.574). Additionally, population structure generated two subpopulations. Group 1 corresponded to Z. japonica from Sumatera Island and mixed province while Group 2 corresponded to Z. japonica from Central Java and Bali with Z. japonica and Z. matrella from mixed province. In conclusion, the exploration of morphological and genetic diversity from Indonesian zoysiagrass provided useful insight for conservation and future breeding improvement
The Survival, Growth, and Accelerating Morphological Development of Stichopus horrens are Affected by the Initial Larval Stocking Densities
Get PDFStichopus horrens is highly exploited due to their use as a pharmaceutical ingredient. Since then, this species has become extinct; therefore, it is necessary to start aquaculture. Gonad maturation and broodstock spawning succeeded, but the optimum larval-rearing stocking density has not yet been determined. Therefore, the present study aimed to determine the optimal stocking density to improve survival and accelerate the development of larval morphology. Three treatments were used: 100, 150, and 200 larvae L-1, each with four replicates. Twelve 100-litre plastic containers filled with 80 litres of seawater as larval rearing media were placed in a concrete tank with a water-bath system using a heater and a thermostat (29.0°C±1.0°C). The larvae were fed a mixture of microalgae (Chaetoceros muelleri, Isochrysis galbana, and Nitzchia sp.) twice a day-1. The data collected included survival and growth rates, larval morphological development, and water quality. The fastest metamorphose from auricularia to the doliolaria stage occurred in 100 larvae L-1, 15 days after hatching. The highest survival rate, growth rate, and percentage of larvae metamorphose to the doliolaria stage were obtained in the 100 larvae L-1 as 26.38%, 26.4 µm day-1, and 65.27%, respectively, and were significantly different (P<0.05). A stocking density of 100 larvae L-1 was optimal for promoting survival and growth and accelerating the morphological development of auricularia to the doliolaria stage
Assessment of Biological Activity, Total Phenolic Content, and Cytotoxicity of Ethyl Acetate Extracts from an Endophytic Fungus, Lasiodiplodia pseudotheobromae IBRL OS-64
Get PDFThe biological activities of ethyl acetate extracts of Lasiodiplodia pseudotheobromae IBRL OS-64 were assessed in the present study. Antioxidant activity was measured using the DPPH scavenging assay, antibacterial activity via broth microdilution, and total phenolic content using the Folin-Ciocalteu method. In vivo toxicity was determined using a brine shrimp lethality assay. The fraction extract exhibited the highest antioxidant activity, with an EC50 of 441.6 µg/ml, surpassing the crude extract\u27s. The EEELP showed a relatively low phenolic content of 13.273 µg GAE/mg extract. Regarding antibacterial activity, both crude and fraction extracts demonstrated MIC values ranging from 62.5 to 250 µg/ml against Gram-positive bacteria, while MBC values ranged from 125 to 500 µg/ml for both extracts. The MIC values for crude and fraction extracts against Gram-negative bacteria were 250-500 µg/ml, whilst the MBC values for both crude and fraction extracts were in the range of 500–2,000 µg/ml. The findings also revealed that Gram-positive bacteria were more susceptible to both extracts (crude and fraction) than Gram-negative bacteria. The crude extract exhibited a non-cytotoxic effect with an LC50 value of 2054.88 µg/ml for acute exposure and a low cytotoxic effect with an LC50 value of 199.69 µg/ml for chronic exposure. Meanwhile, the fraction extract demonstrated a non-cytotoxic effect for both acute and chronic with an LC50 value of 5744.14 µg/ml and 1035.94 µg/ml, respectively. Therefore, the present study suggests that the fraction extract could be an effective pharmaceutical agent and safe for drug development due to its low toxicity
Increased Anti-Proliferation Performance of NanoChitosan-Moringa Seeds Extract and Co-Treatment with Doxorubicin in Liver Cancer Cells
Get PDFHepatocellular carcinoma (HCC) with high epidemiological report data. Pathogenesis in HCC also involves several signaling pathways. This study aims to evaluate the in vitro activity of Moringa seed NanoChitosan against Hep G2 liver cancer cells and Co-Treatment with Doxorubicin. Initially, nanoparticles were prepared by extracting Moringa seeds, formulating them into nano chitosan, and then characterizing the compounds and particle sizes. The IC50 dose was investigated using the MTT assay. Then, the IC50 dose was confirmed in more detail through immunofluorescence, betatrophin gene, several genes in the Wnt-βcatenin-CyclinD1 proliferation pathway, and the addition of the apoptotic effector Caspase-3 using RT-qPCR analysis. Each treatment used a single dose of NCH-Mosee and co-treatment or combination with 4 μg/ml doxorubicin. The IC50 dose was 994 μg/ml in single treatment and 649 μg/ml in combined treatment with Dox. Hep G2 showed a decrease in the expression level of each parameter measured with increasing single dose and combination treatment (p < 0.050). Histologically, cells shrank, betatrophin expression was inhibited, and luminescence was seen, which decreased with increasing dose. In conclusion, NCH-Mosee with dose-tracking toxicity combined with Dox can suppress the viability of Hep G2 cells