HAYATI Journal of Biosciences
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Isolation of DNA Aptamers for Enteropathogenic Escherichia coli (EPEC) Detection using Bacterial-SELEX Approach
Get PDFEnteropathogenic Escherichia coli (EPEC) is a Gram-negative pathogenic bacterium that causes diarrheal disease, especially in infants and children. Aptamers are short chain oligonucleotides that have high affinity, specificity, and selectivity to their targets, which have potential to be developed as a method for diagnosing pathogens. In this study, aptamer was isolated through the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method using whole cells bacteria (Bacterial-SELEX) for recognizing pathogenic E. coli EPEC K1.1 which was isolated from children with diarrhea in Indonesia. Ten rounds of bacterial-SELEX procedure were conducted with modification conditions by using Top10, DH5a E. coli cells, Listeria monocytogenes, and Lactobacillus plantarum S34 as counter-selections. The selection process was started with a pool of ssDNA random library consisting of a random base with 40-nucleotides long flanked with fixed primers sequence for aptamer amplification purpose. Short single-stranded DNA amplification was done by symmetric and asymmetric PCR. The highly enriched oligonucleotide pools (pooled 8, 9, and 10) were cloned and the resulting ssDNA aptamers were identified by Sanger DNA sequencing. Finally, twelve aptamers with unique sequences and various secondary structures including G-quadruplex sequence motif within aptamers were obtained as candidates specific aptamer for detection and capturing of EPEC K1.1
A Meta-Analysis of Latex Physiology Studies Reveals Limited Adoption and Difficulties to Interpret Some Latex Diagnosis Parameters in Hevea brasiliensis
Get PDFLatex diagnosis is widely adopted in natural rubber-producing countries to optimize the natural rubber production through a physiological-based latex-harvesting system management. This study is the first bibliographical searching and meta-analysis on the variation of latex physiological parameters i.e. sucrose, inorganic phosphorus, thiols, and total solid content. The study used information extracted from 158 scientific papers. Descriptive statistics, agglomerative hierarchical clustering, and principal component analysis were performed to characterize applications of latex diagnosis, how often parameters are used and interpreted as well as the variation of its parameter values. From 158 papers, latex diagnosis parameters were used in 114 agronomy, 22 in physiology, and 22 in breeding papers. The agglomerative hierarchical clustering analysis indicated that sucrose and inorganic phosphorus contents were clustered together and total solid and thiols contents were located in another cluster. The average values of the total solid content, sucrose, inorganic phosphorus and thiols were 43.9%, 9.4 mM, 16.5 mM, and 0.52 mM, respectively. The percentage of interpretation is 63.3% for sucrose, 54.4% for inorganic phosphorus, 47.5% for thiols, and 41.1% for the total solid content. The low interpretation of thiols and total solid contents question their relevance in latex diagnosis. The low adoption of latex diagnosis in breeding could hinder the selection of activities leads to a limitation of selection for long-term high yielding and stress-adapted clones
Genotype-Environment Interaction and Yield Stability of Upland Rice in Intercropping Cultivation
Get PDFRice cultivation as intercropping in plantations is a potential approach to increase land resource efficiency. Implementation of rice intercropping would require genotypes adapted to shaded environments. This study aimed to clarify the interaction of genotype and environment on agronomic characteristics of upland rice grown under intercropping system and to identify suitable genotypes for this particular cropping system. Multi-location trials evaluated twelve upland rice lines and two check varieties in eight upland sites during the wet season (WS) 2015-2016. Rice was grown as intercropping in six sites and as monoculture in two sites. Rice as intercrop was cultivated in between plantations of tall coconut, rambutan, teak, albizia, and natural rubber. The result of this study revealed that the interaction between genotype and environment was significant for all agronomic characteristics. The genotype B12056F-TB-1-29-1 was the best performer when grown as intercropping in Subang and Indramayu, with shading intensities of 65% and 44%, respectively. The genotype B11908F-TB-3-WN-1 was the best yielder in Cianjur and Lampung Tengah, which had 37% and 54% shading intensity, respectively. The genotype B12825E-TB-2-4 was the best performer in Lampung Timur with shading intensity of 45%, while Jatiluhur was the best genotype in Banyumas with shading intensity of 82%. The yield of rice genotypes grown as monocultures was higher than as intercropping. Among the genotypes, B12056F-TB-1-29-1 showed the highest yield across sites. Stable and high-yield genotypes identified from this study are expected to be adopted by farmers for intercropping rice cultivation
Reef Fish Biodiversity at Different Depths in Tunda Island, Banten Province, Indonesia
Get PDFReef composition and diversity of coral reef fishes play an essential role in the ecosystem. The reef fish diversity is a crucial indicator of the health level of coral reef ecosystems. The benefit of fish diversity in coral reef ecosystems is to describe ecological services. This research intended to determine reef fish diversity among stations at different depths, consisting of shallow water and reef slope. Research sampling was done in July 2018 and July 2019 in Tunda Island waters at eight stations billowing the coral distribution pattern. Collected data of reef fishes were by using Underwater Visual Census (UVC) on Belt Transects Method at 250 m2 transect area (50 meters of length and 2.5 m on either side). The result showed 69 reef fishes in shallow water and 67 species on the reef slope. The total of fish species ware from 13 families consisting of Acanthuridae, Apogonidae, Blenniidae, Chaetodontidae, Ephippidae, Haemulidae, Labridae, Lutjanidae, Nemipteridae, Pomacentridae, Pseudochromidae, Scaridae and Serranidae. The community services structure of coral reef fishes is included in the moderate community stability category with an H’ value ranging from 1.19-1.60 in shallow water and 1.23-1.44 at reef slope, respectively. Diversity in coral reef fish communities might be associated with species\u27 spatial distribution over the reef habitat\u27s physical structure, but these patterns could be highly scale-dependent. In general, it could be said that the higher the coral reef diversity, the higher the reef fish diversity
Role of Fe2+-dependent Reaction in Biodecolorization of Methyl Orange by Brown-rot Fungus Fomitopsis pinicola
Get PDFThe involvement of Fenton reaction on biodegradation of methyl orange (MO) by brown-rot fungus Fomitopsis pinicola was investigated based on Fe2+-dependent reaction. The degradation of MO (final concentration 75 mg/L) was performed in mineral salt media with and without Fe2+ with incubation period at 0, 7, 14, 21, and 28 days. Degradation analysis was performed using UV-Vis Spectrophotometer and LC-TOF/MS. F. pinicola decolorized MO in a medium containing Fe2+ and a medium that lacked the mineral, at percentages of 89.47% and 80.08%, respectively. The optimum decolorization occurred after 28 days of incubation with the fungus on the presence of Fe2+, indicated that the presence of Fe2+ enhanced MO degradation with assumed to correlate with Fenton reaction. Two metabolites were detected through the LC-TOF/MS analysis, namely 4-(2-(4-(dimethyliminio)-2-hydroxycyclohexa-2,5-dien-1-ylidene) hydrazinyl) phenolate (m/z 258, RT: 1.28 min, compound 1) and 4-(2-(4-(dimethyliminio) cyclohexa-2,5-dien-1-ylidene) hydrazinyl) benzenesulfonate (m/z 391, RT: 2.70 min, compound 2). Compound 1 was a transformation product of hydroxylation and methylation, compound 2 was a product of dehydroxylation and desulfonation. This study indicated that the transformation of the metabolite structures was involved hydroxyl radical (OH.) and enzymatic mechanisms, which involved Fe2+-dependent reaction
Cross-Species Amplification and Variability of Microsatellite DNA Markers in Domesticated Indonesian Mahseer; A Case Study with Tor soro, Tor douronensis and their Interspecific Hybrids
Get PDFIndonesian mahseer (Tor spp.) are freshwater species of high economic, cultural, and conservatory values. Owing to their high values and environmental degradation, the population of Tor fish gradually decreased, and domestication efforts have been made to conserve the population. This study was aimed to assess the cross-species amplification and microsatellite genetic diversity in Indonesian mahseer Tor soro (SS), Tor douronensis (DD), and their interspecific hybrids using primers developed for Tor putitora. Eleven primer sets were used to test for amplifiability and screen genetic diversity in 40 progenies derived from those three groups. Results showed that seven primer sets (64%) successfully amplified loci. Genetic screening using the three most consistently amplifying primers showed that the number of alleles in the three populations was low, ranging from 2 to 5 alleles. The observed heterozygosity (HO) was high ranging from 0.650 to 0.789, and the fixation index (FIS) was negative, indicating heterozygosity excess. In line with other parameters, the P-values of the HW parameter of several loci-population combinations were significantly departed from equilibrium (P <0.05). A few private alleles were observed in parental line DD and the hybrids. Overall, the cross-species primers developed from T. putitora were able to amplify loci in T. soro, T. douronensis and their hybrids and genetic diversity in the hybrid population was slightly higher than those in parental lines. Possible factors driving the phenomena and practical implications of these findings on the conservation measures are discussed
Molecular Detection of Hemagglutinin Gene Fragment of Avian Influenza Virus H9 Subtype Obtained from Poultry Commercial Farm with Prominent Symptom of Decreased Egg Production
Get PDFAvian influenza virus H9N2 subtype is considered a low pathogenic strain that has been reported in Indonesia since late 2016. The outbreak has caused economic losses for farmers due to the sharp drop in egg production. The evidence of the existence of AIV H9N2 has been published, but very limited information on the prominent symptom and macroscopic lesion. This research was a retrospective study of suspected avian influenza H9 subtype, obtained from layer commercial farm with recorded characteristic symptoms. Specific trachea samples were collected and further processed to be isolated, propagated using embryonated chicken egg, and then extracted the RNA for molecular detection using real-time reverse transcriptase-polymerase chain reaction (RT-qPCR). A positive result was further detected in its H-9 gene with RT-PCR technique and sequencing methods. One of five samples showed positive for RT-qPCR with CT value 30.19. Sequence analysis confirmed that the sample with characteristic macroscopic lesion could be detected in the presence of the AIV H9 subtype. Phylogenetic tree analyses revealed that this virus belongs to the China-Vietnam- Indonesia (CVI) lineage
Endophytic Fungi Isolated from the Mangrove Species Rhizophora apiculata and Their Efficacy as Herbicides
Get PDFEndophytic fungi play an important role on mangrove growth and development, however research on the endophytic fungi of the mangrove Rhizophora apiculata is limited. The endophytic fungi produce diverse bioactive compounds involved in the mangrove’s adaptation to varied biotic and abiotic stresses and could have applied uses in agriculture or medicine. The purpose of this research was to isolate endophytic fungi of Rhizophora apiculata mangrove and to study their activity as herbicides on the weed Gomphrena globosa. The fungi were isolated using the surface sterilization method and identified based on morphological characteristics and molecular characteristics using ITS regions of rDNA. Herbicidal activity of the fungal filtrates extracted by ethyl acetate were tested on seed germination and seedling growth of G. globosa. Five fungal isolates were obtained, namely Penicillium citrinum, Diaporthe eucalyptorum, Diaporthe musigena, Colletotrichum queenslandicum, and Diaporthe tectonae. All isolates were able to grow on PDA medium containing 0, 25, 50, 75, and 100 (% v/v) seawater concentrations, but the growth rate varied by species and seawater concentration. In general, all five isolates showed herbicidal activity by delaying seed germination and reducing shoot and root growth. P. citrinum showed the highest herbicidal activity compared to the other isolates. Analysis using Gas Chromatography-Mass Spectrometry of the crude extract of P. citrinum filtrate identified 7 main compounds: 3-Methoxy-2-methyl-cyclohex-2-enone, Cyclohexane-carbohexaldehyde, 6-methyl-3-(1-methylethyl)-2-oxo, Cyclopropane carboxylic acid, 1-(2-propenyl)-1,1-dimethylethyl ester, 2-Hydroxy-4-isopropyl-7-methoxytropone, Beta-Asarone, Oxane, 4-(2-amino-1-hydroxyethyl)-4-(3,4-dimethoxyphenyl)-, and Diisooctyl-phthalate. These compounds should be studied further to determine which ones are responsible for the herbicidal activity
Genetic Background of β-Lactamase Genes in Extraintestinal Pathogenic Escherichia coli ST131 in Indonesia Based on Whole-Genome Sequencing (WGS) Sequences
Get PDFExtraintestinal pathogenic Escherichia coli (ExPEC) is a group of pathogens that can colonize the outside of the intestine, such as the kidney, urinary tract, or bloodstream. This study analyzed more about the genetic background of β-lactamase genes among Indonesian ExPEC ST131. Whole-genome sequencing (WGS) sequences from the National Center for Biotechnology Information (NCBI) of Indonesian ExPEC ST131 were taken, then analyzed. Circular genomic mapping and genomic comparison of surrounding genes of β-lactamase in these isolates were generated. blaOXA-1 and blaDHA-1 were analyzed deeply due to their presence in a relatively long contig, making them available for analysis. Indonesian ExPEC ST131 isolates had blaOXA-1 with an identical genetic background of E. coli originating from China and Austria with aac (6’)-1b-cr5 in the downstream and cab83 in the upstream. The blaOXA-1 was also detected in other species, including Klebsiella pneumoniae INF142 originating from Australia and Salmonella enterica S146 from China. While, blaDHA-1 in EC_0406 had an identical genetic background to E. coli 142 and other species such as Shigella sonei FC1428 from India and S. flexneri M2901 from Australia, with Globulin-encoded gene in the upstream and lysR in the downstream. Our findings demonstrate the global spread of β-lactamase genes detected in Indonesian ExPEC ST131. These genes were identical to those isolated from some countries around the world
Molecular Identification and Phylogenetic Analysis of Fungi Contaminants Associated with In Vitro Cultured Banana Based on ITS Region Sequence
Get PDFThis study characterized, identified and conducted phylogenetic analysis on fungi contaminants in vitro bananas based on the sequence of inter-space (ITS) regions. Genomic DNA was extracted from the pure culture of fungi contaminants, amplified and sequenced using ITS1 and ITS4 markers. Analysis of the sequences using MEGA 7 Software at higher similarity sequence identified five Aspergillus spp., three Penicillium spp., one each of Fusarium, Trichoderma and Cladosporium as the contaminants. The genetic distance between the fungi species was 0.205, which suggests a homogeneous substitution between the sequences, and thiamine was the most stable. The fungi clustered in three major groups at 0.10 genetic distance, subdivided into five clusters. A cluster and sub-cluster consisting of five Aspergillus strains; a major cluster of three Penicillium strains; a cluster comprising of Fusarium chlamydosporum and Trichoderma viride; and a sole fungi Cladosporium tenuissimum. The Aspergillus group were phylogenetically related to A. flavus and A. parvissclerotigenus, the identified Penicillium spp. were closely related to Penicillium citrinum while the detected Cladosporium aligned with Cladosporium tenuissium and Phoma multirostrata. The information provided by this study could be utilized to develop a specific and compelling sterilization protocol to minimize the rate of contamination during in vitro culture procedures