Sanger Sequencing Customer Publications

Tailing from mining activities affects soil fertility resulting in poor soil conditions that are challenging for plants to grow. Plants can interact with rhizosphere bacteria to enhance their growth in harsh environments. Rhizospheric bacteria possess numerous mechanisms that promote plant growth and induced resistance to various abiotic stress. This study aims to determine the diversity of rhizobacteria and their potential as plant growth-promoting rhizobacteria (PGPR) agents. Bacterial communities from rhizosphere soil samples from kaolin mining sites in Perawas, Tanjung Pandan district, Belitung Regency, Bangka Belitung Island, Indonesia were analyzed using Next Generation Sequencing based on the V3-V4 region of the 16S rRNA gene, while culturable bacteria were isolated from samples and screened for PGP activity. The results showed that the rhizosphere bacterial community was mostly dominated by Pseudomonadota, Acidobacteria, and Verrumicrobiota. There were 15 bacteria isolated from the sample and RKB-5 bacterial isolate had the potential to be PGP agent. The RKB-5 bacterial isolate was identified as Burkholderia cenocepacia based on its 16S rRNA sequence. The bacterial isolate produced IAA, utilized ACC, dissolved phosphate up to 209,5 mg/L, and formed a high potassium solubilizer index value of 5.00. Therefore, the B. cenocepacia RKB-5 has potential application as the PGPR to support plants growth by obtaining nutrients in ex-mining lands with poor soil conditions.

This study aimed to analyze the diversity of indigenous bacteria by comparing culture and non-culture methods and to analyze the physicochemical effects on bacterial diversity in polluted and natural mangrove sediments. The environmental parameter values of mangrove sediments for bacterial growth can change owing to differences in adaptation and tolerance to fluctuations in physicochemical conditions. The number of colonies in natural and polluted areas using the culture method was 6.2 × 104 CFU/g and 5.5 × 104 CFU/g, respectively. A total of 33 isolates were identified, with 17 and 16 isolates from the natural and polluted areas, respectively. The most common isolates found in both areas were Acinetobacter haemolyticus strain FBC636 and Exiguobacterium acetylicum strain IAE17. Using the nanopore sequencing method, the total number of colonies in the natural and polluted areas was 69,761 and 58,412 colonies, respectively. A total of 12,954 bacterial species were identified, with 6,837 species in the natural area and 6,117 in the polluted area. The most common isolate found was Sulfurovum aggregans. Physicochemical conditions influenced the differences in bacterial diversity between the natural and polluted areas in the mangrove areas of Ambon Bay.

Cloning and expression of RubiscoLike Protein (RLP) from halophilic bacterium Chromohalobacter salexigensBKL 5.Biodiversitas 25:1605-1614.This research seeks to identify and determine the Rubisco-like proteincharacteristics from Chromohalobacter salexigensBKL 5 tested using a recombinant method approach. C. salexigensBKL 5 is a halophilic bacterium found in Bledug Kuwu Mud, Central Java. The Open Reading Frame (ORF) gene encoding Rubisco-likeprotein from that bacterium was successfully amplified using the polymerase chain reaction method using the primers we designed. Cloning of that gene on the pCold plasmid was carried out using the double digest method with the restriction enzymes NdeI and EcoRI.The results of plasmid transformation in Escherichia coli BL 21 were successful, indicating that the bacterial culture was grown in Luria Bertoni agar media containing the antibiotic ampicillin. Overexpression protein of the transformed Escherichia coli BL 21 shows that the effective temperature that produces maximum results is 16°C. The results of sequencing and phylogenetic analysis showed that the protein belongs to the Rubiscofamily, namely Rubisco-likeprotein with a residue of 429 amino acids and a weight of 46 kDa. Prediction of 3D structures using the Alphafold tool givesan accuracy above 90%. Observation of the 3D structure shows that glutamic acid is more distributed on the surface of the proteinwhich is important for protein solubility. Tests with SOPMA showed that this protein was dominated by random coil structure of 41.96% which was identical to that found in bacterial species in extreme habitats. The dynamic molecular analysis using Yasara at 10 ns showed that the protein had a tendency to undergo considerable deformation but remained stable.

Geosesarma shows intraspecific carapace color variation, which might lead to species misidentification. The problem can be solved using DNA barcoding. There is one research about Geoserarma from the southern slopes of Mount Slamet, but samples were only collected from the Banjaran River for morphological identification. Here, we collected samples from wider areas covering south slope and applied molecular identification. This research aims to assess Geosesarma diversity in south-slope Mount Slamet Central Java, Indonesia based on the cytochrome c oxidase 1 gene barcoding. Surveys were carried out at six sites. Taxonomic identification was done using the barcoding technique. Four morphotypes were obtained during the research. Three morphotypes with the square carapace were identified as Geosesarma, while the remaining one morphotype was included in Parathelphusa. The three Geosesarma morphotypes were barcoded as Geosersarma dennerle because their genetic identity was more than 97% of the G. dennerle sequence in Boldsystems. In contrast, the Parathelphusa morphotype was barcoded as P. convexa with a genetic identity of 97.50%. It can be concluded that the Geosesarma crab on the south-slope Mount Slamet only consists of one species but has carapace and claw color variations. The data are essential for Geosesarma market development and conservation in the region.

Cardamom is one of the prominent plants with significant economic value in the Banyumas Regency, Central Java, Indonesia. Although Javanese cardamom is traditionally categorized under Amomum compactum, the morphological variations observed create ambiguity about its exact species status. DNA barcoding using the maturase K (matK) and ribulose-bisphosphate carboxylase (rbcL) genes is proven as a reliable technique to elucidate the taxonomic status of morphological variable plant cultivars. This study aimed to characterize cardamom from Banyumas Regency using morphological and molecular approaches for taxonomic status identification and genetic diversity evaluation. The matK and rbcLgenes were selected as genetic markers andsequenced using a bidirectional sequencing technique. Morphological examination showed significant color variations at the cardamom stem base. All samples had high genetic identities to reference species in databases and were supported by high query cover and zero e-values. Therefore, molecular characterization, alongside geographic distribution assessment, established that this plant belongs to a single species, Amomum compactum. Additionally, the analysis conducted showed a low level of genetic diversity, as evidenced by haplotype and nucleotide diversity. Low-levelgenetic diversity provides additional data to convince that cardamon in Banyumas Regency belongsto a single species. These results are essential data in seed selection for further cultivation.

Aeromonas spp. causes the human diseases including diarrhea, gastroenteritis, and bacteremia. Aeromonas spp. can be found in kitchen sponge, one of the reservoirs for food-borne bacterial pathogens. Virulence study of Aeromonas spp. in vivoin animal model is important since the animal model can mimic manifestasions in human infections. Omphisa fuscidentalis was chosen for alternative virulence model, since they are in the same taxonomical order with the well-known infections model, Galleria mellonella. Bacterial isolation and selection of kitchen sponge used Brain Heart Infusion agar and Endo Agar, respectively. Bacterial virulence of KS-1 was injected into Omphisa fuscidentalis larvae. Survival percentage and melanization score of infected larvae were evaluated. Hemolymph of larvae with melanization score of 1 and 4 were stained with Giemsa method to observe the hemocyte changes. Bacterial identification of isolate KS-1 based on 16S rRNA sequence resulted in 96.9% identity to Aeromonas spp. strain VS7. Isolate KS-1 injection to O. fuscidentalis revealed higher bacterial dosage resulting more severe symptoms to the larvae according to survival percentage and melanization score. However, statistical analysis showed evaluation of melanization score could distinguish larvae with 106 and 107 CFU/larva dosage injection, while evaluation of survival percentage could not. Hemocyte of larvae with melanization score 1 had larger and more cytoplasmic vacuolization than the score 4 (healthy larvae). Omphisa fuscidentalis is an alternative of insect model for bacterial infections with survival percentage and melanization score as the evaluation. Cytoplasmic vacuolization of hemocyte can be used as larvae’s health indicator in a cellular level

Infestation and histopathology of Clinostomumsp. (Digenea: Clinostomidae) in snakeskin fish (Trichopodus pectoralisRegan, 1910) in Tempe Lake, South Sulawesi, Indonesia.Biodiversitas 25:3263-3272.Fish diseases caused by parasites seriously threaten the aquaculture industry and wildfish populations in natural water. This research aims to identify and characterize the digenetic trematode Clinostomum sp. based on morphology and molecular characteristics and analyze the level of infestation and histopathological changes of the endoparasite infestation in snakeskin fish across three lakes (Tempe Lake, Lampulung and Latamperu Lakes) in South Sulawesi, Indonesia. A total of 300 snakeskin fish were collected from a survey conducted from July 2023 to January 2024 for parasite examination. The morphological identification of parasites, a meticulous process, was carried out to distinguish digenean species that infect snakeskin fish populations, ensuring the accuracy of our findings. The results showed that Clinostomumsp., was found infesting snakeskin fish across all three lakes within Wajo District, and the identity of the parasite was further confirmed with Polymerase Chain Reaction (PCR) and sequencing on the Internal Transcribed Spacer (ITS)-5.8S region. The prevalence and mean intensity ofthe parasite infestation varied among the three locations surveyed, though they are not different statistically; Lampulung Lake showed a prevalence of 34%, Latamperu Lakeat 28%, and Tempe Lake at 21%. The mean intensity of infestation in Lampulung Lake at 10±10.70, Latamperu Lake at 09±12.82, and Tempe Lake at 06±6.22. Histopathological analysis showed changes within the intestinal tissues of snakeskin fish associated with the infestation of Clinostomum sp., including goblet cells, hemorrhage, and cellular infiltration. This study demonstrated a consistent presence of Clinostomum sp. across all lakes, with relatively high prevalence and mean intensity of infestation. The histopathological analysis showed mild tissue alterations in infested fish. However, the high prevalence of infestation of the zoonotic digenetic trematode raises concerns about the public health impacts for local communities who consume raw or undercooked fish.

 DNA barcoding of intertidal barnacles as potential bioindicators of microplastic pollution in Seribu Islands and Jakarta Bay, Indonesia. Biodiversitas 25:4664-4676.Barnacles, with their sessile nature and filter-feeding behavior, hold significant potential as bioindicators of microplastic pollution. Due to the diverse morphotypes across barnacle species, DNA barcoding is a reliable technique for accurate taxonomic identification. This research aimed to determine intertidal barnacle species and identify potential bioindicators of microplastic pollution in Seribu Islands and Jakarta Bay,Indonesia. Barnacle samples were collected from seven locations using purposive sampling. Microplastic characteristics were analyzed visually and polymer-type testing was performed using Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (ATR-FTIR). Data was analyzed using Pearson correlation and bioconcentration factors to determine potential microplastic bioindicator species based on three criteria. The significance level was set at p<0.05 and all statistical analyses were performed using SPSS. Four species of intertidal barnacles were identified in Seribu Islands and Jakarta Bay, namely Amphibalanus amphitrite, Striatobalanus amaryllis, Amphibalanus zhujiangensis, and Newmanella radiata. DNA barcoding was used to determine the first three species, while morphological analysis identified the fourth species. The microplastic particle count varied among the species, with A. amphitriteshowing the highest concentration at 42-53 particles/g. Due to its clear taxonomy, ease of surveying, and wide distribution, A. amphitritehas strong potential as a bioindicator of microplastic pollution, as it can accumulate more than other species

Natural association of the entomopathogenic fungi Aschersonia placentawith spiralling whitefly (Aleurodicus dugesii) in Bali, Indonesia. Biodiversitas 25:4067–4073.The whitefly is one of the most common pests in agriculture,and one highly polyphagous species is the spiral whitefly (Aleurodicussp.). The fungal genus Aschersoniacan be used to control Aleurodicussp..This research has potential applications in agriculture because it can produce a new environmentally friendly method to control whitefly infestations. The aim of this study was to analyze the morphology and molecular characteristics of Aschersoniaand their hosts in Bali. The samples were taken from several areas in Bali Province. Morphological identification was carried out at the Plant Pathology Laboratory, Faculty of Agriculture, UniversitasUdayana. The conidia of Aschersoniawere fusoid with tapered ends, 9–16 μm long and 1.5–2 μm wide,and the color of the culture was white to yellowish white. Molecular analysis exhibited that DNA bands measuring between 500–600 bp were successfully amplified with universal primers ITS1/ITS4. Aschersoniainfected spiralling whiteflies has never been reported in mulberry plants. Further phylogenetic analysis showed that Aschersonia–infected spiralling whiteflies were in the same group as Aschersonia placenta/Hypocrella raciborskiiisolates from Thailand and India. Aleurodicus dugesii Cockerell 1896had morphological characteristics, such as vasiform orifice with lingula extending beyond the borders of orifice, compound pores present on puparia, and thoracic legs with claws. The results of morphological analysis showed that A. dugesiifrom Bali (LC491422) had the closest kinship to A. dugesiifrom the USA (AY521251). This is the first report of the identification of Aschersoniaplacenta associated with A. dugesiiin Indonesia.

Mitochondrial DNA of CO1 and 16S rRNA genes for diversity and phylogenetic analysis of sardine (Sardinellaspp.) from the Lombok Straits, Indonesia. Biodiversitas 25: 3500-3509.Sardines in Indonesia consist of six species from around 21 species-world, having many similarities and difficulty distinguishing between one species and others caused the taxonomy of the Sardines species is still ambiguous affected the same names in two different species and the different names in one species because of morphological similarities. The most effective method for identifying animal species and subspecies is themolecular method using sequences of the cytochrome c oxidase subunit 1(CO1) gene and the 16S rRNA gene. The study aims to determine the molecular characteristics, genetic diversity, and phylogenetic relationships of sardines in the Lombok Strait using a sequence of CO1 and 16S rRNA genes. The study has significant practicalimplications as it can potentially inform conservation efforts and fisheries management. The sardine samples were collected from the Lombok Strait by small-scale fishermen. The sampling method was random. It takes 55 individuals as a sample. Initially, the sardine samples were grouped based on morphological characteristics. Subsequently, DNA from the two genes of 20 sardine samples were separated and amplified using two pairs of primers. The DNA amplification yielded 671 bp for the CO1 gene and 625 bp for the 16S rRNA gene. The sequence data from the two genes were analyzed using Molecular Evolutionary Genetics Analysis Version 11 (MEGA 11). The CO1 gene sequence revealed that sardines from the Lombok Strait could be determined more accurately than the 16S rRNA. CO1 rRNA genes of 12 individuals indicated a close genetic relationship between sardines of Lombok Strait and Sardinella aurita Valenciennes 1847, and the 16S rRNA gene placed sardines from Lombok Strait as an out-group. It empirically demonstrates that the CO1 gene is more effective than the 16S rRNA gene in identifying and determining molecular characteristics, genetic diversity, and phylogenetic relationships of sardines in Lombok Strait.

Threat of extinction of Macrobrachium esculentum in the Serayu River (Central Java, Indonesia) confirmed by DNA barcoding. Biodiversitas 25: 3531-3539. Sweet river prawn (Macrobrachium esculentum Thallwitz 1891 is one of the amphidromous shrimp. Serayu River is one of the habitats of this shrimp. Information about the M. esculentum in the Serayu River, is still minimal, so this research is essential. The Serayu Weir affects the population of Macrobrachium esculentum, which impacts the balance of the Serayu River ecosystem. This study aims to determine the condition of the M. esculentum population in the lower reaches of the Serayu River in order to maintain its sustainability and avoid the threat of extinction. This research was conducted for one year (January-December 2023). The method used was the descriptive method. We collected M. esculentum catches both upstream and downstream of the weir for one year to determine the presence of shrimp as a result of the Serayu Weir. The catch data was compared with the catch data from previous research. M. esculentum has morphological characteristics with a blackish-gray pattern with stripes along its abdomen and has an upper rostrum of 11-14 teeth and 2-4 on the lower rostrum. M. esculentum in the Serayu River has an mtDNA fragment of 686 bp and shows the same species as M. esculentum in GenBank. A total of 65 M. esculentum were collected. This shrimp was not found above the weir (Station 1) but only at station 2 (below the weir) due to the presence of weir. The results showed the importance of constructing migration routes for aquatic biota in the Serayu Weir. Further research is also needed, especially related to the reproductive system of M. esculentum.

The occurrence of the very rare species Gekko cf. brooksii (Squamata, Gekkonidae) in West Sumatra, Indonesia, based on molecular and morphological evidence. Biodiversitas 25: 3369-3379. Sumatra, Indonesia is a region of vast, rich and diverse flora and fauna, yet knowledge of herpetology in Sumatra is limited. One such group is the genus Gekko, which is poorly known, morphologically diverse and taxonomically problematic, especially for canopy-dwelling geckos. During our recent fieldwork in the Barisan Mountains of West Sumatra, we found an unidentified camouflaged gecko and used molecular and morphological approaches to confirm its identity. Based on molecular (NADH dehydrogenase 2 [ND2] and flanking tRNA genes) and morphological comparisons (31 characters), we confirmed a new record and an expansion range of Gekko cf. brooksii to West Sumatra; we also reported several morphological characters that have not been reported in previous studies. The individual was found on the leaves of a young breadfruit plant (Artocarpus communis J.R.Forst. & G.Forst.) located about 1 meter from the hiking trail, where the surrounding vegetation consisted of several tall trees and was mostly dominated by herbaceous plants that did not exceed 1 meter in height. It conclusion, despite a long history of exploration, the herpetofauna of Sumatra continues to yield new discoveries and records. As the geckos inhabiting the higher canopy layers of tropical rainforests in Sumatra remain largely unknown, further intensive surveys for canopy-dwelling geckos are needed to further elucidate the complete taxonomic composition of Sumatra’s herpetofauna.

New specific primer matK and rbcL region for DNA barcode pitcher plant Nepenthes spathulata. Biodiversitas 25: 2515-2523. The matK and rbcL genes were proposed as the preferred plant barcoding loci by The Consortium for the Barcode of Life (CBOL). DNA barcoding efficiently identifies samples at the species level using short standard DNA sequences. The identification disclosure needs is an important basis given the confusion in systematics in Nepenthes spathulata. Therefore, this study aims to design matK and rbcL region-specific primers for DNA barcoding pitcher plants. The primary design uses Primer3Plus with a blueprint from the GenBank KDQ007081.1 for matK and MH346374.1 for rbcL. Following testing primary candidates with Oligo Analyzer and Primer-BLAST, the primers MkNs1, and RbNs4 were selected as the optimal criteria based on the values of ?G and primer specificity. Optimization of annealing temperature with PCR gradient shows that the temperature range 52-57°C produces a good band for both regions and corresponds to the matK and rbcL region with product sizes of 800-900 bp from seven samples, which could be observed from the gel electrophoresis. Sequence similarity using blast-N, the matK sequence has similarity to the N. spathulata matK gene of 99.64% and 99.63% for the rbcL gene of N. ventricosa x N. alata. This MkNs1 and RbNs4 primer can be used to discover the identity of N. spathulata and the Nepenthes genus.

Hidayati N, Soffan A, Arwiyanto T, Wijonarko A. 2024. Diversity of bacterial endosymbionts of Bemisia tabacifrom some regions in Indonesia and the genetic diversity of Wolbachiaendosymbiont. Biodiversitas 25: 4304-4314.Bemisia tabaci Gennadius 1889(Hemiptera) is an insect that associates with endosymbiont bacteria to meet nutritional needs lacking in its food. The species has both primary and secondary endosymbionts, with infections showing significant dynamism among populations.Wolbachiais a facultativeendosymbiont that infects 66%of all insect species, including B. tabaci,which is a key focus of this study. The study aimedto investigate the diversity of bacterial endosymbiont infections in several B. tabacipopulations collected from Java and Sumatra, Indonesia, to explore the genetic diversity of Wolbachiaendosymbiont.The B. tabacipopulation was collected from 17 districtsin Java and Sumatra, and to determine the presence of bacterial endosymbiont was determined through molecular analysis using specific primers. The study involved a comprehensive methodology, including the construction and comparison of multiple alignment sequences, phylogenetic analysis, and genetic differentiation of Wolbachiain 10 representative populations of B. tabaci, which were then compared with Wolbachiasequences from other countries and arthropods. The results showed the presence of various bacterial endosymbionts found among the B. tabacipopulations, including Candidatus portiera, Arsenophonus sp.,Cardinium sp.,Hamiltonella sp.,Wolbachia sp.,andRickettsia sp. Wolbachia was found in all B. tabacisamples, while other endosymbiont bacteria varied quite widely among all B. tabacipopulation samples.The genetic diversity of Wolbachia of B. tabacifrom Indonesia showed close relatedness and has adifferent clade from Wolbachiafrom other countries and arthropods. The population structure of Wolbachia populations from Java was genetically related and similar to that of Wolbachia from Sumatra.

SetyaningrumN, LestariW, NuryantoA. 2024.Small scale genetic structure of striped snakehead, Channa striatain the river and swampy areas of the south region of Central Java, Indonesia.Biodiversitas 25:2959-2966.This study focuses on the exploration and characterization of Channastriatapopulation in swampy areas located in south-central Java, Indonesia, specifically in the regencies of Purworejo, Kebumen, Banyumas, and Cilacap. These regions are known for their diverse and unique ecosystems, making them ideal sites for ecological research. Fragmented populations of striped snakehead (Channa striata)are observed in the aquatic ecosystems of south-central Java, Indonesia. However, research on C. striatain swampy areas of south-central Java has not been conducted yet. Evaluating the genetic structure of C. striatais a crucial endeavor that can be accomplished through the analysis of the cytochrome c oxidase 1 gene. Therefore, this research aims to estimate the genetic diversity and differentiation among C. striatapopulations. This study analyzed 74 specimensof C. striatacollected from Keburuan, Karangbolong, Jatijajar, Sumpiuh, and Kedungreja in south-central Java. The used marker has moderate haplotype diversity (0.541±0.065) but low nucleotide diversity (0.0025±0.0017). Haplotype diversity within the population ranges from low (0.151±0.093) to high (0.833±0.222), while all populations showed low nucleotide diversity (0.0006±0.0008to 0.0037±0.0030). Through the analysis of genetic markers, C. striatapopulations can be categorized into three distinct groups. The findings revealed that population fragmentation has resulted in reduced genetic diversity and localized population structuring in the river and swampy areas of south-central Java. These results highlight the importance of separate management strategies for each population to ensure their conservation and sustainable management.

Thermophilic bacteria living in extreme areas with high temperatures are capable of producing secondary metabolites, such as antimicrobial peptides (AMPs). AMPs are stable at high temperatures and show good antibacterial activity. Therefore, this study aimed to identify thermophilic bacteria from the crater of Mount Tangkuban Perahu around West Java and assess antibacterial effectiveness of AMPs against Streptococcus mutans, which contribute to oral biofilm formation. The isolate obtained was identified using 16S ribosomal ribonucleic acid (rRNA) gene sequencing, and the supernatant of the isolate was tested against S. mutans American Type Culture Collection (ATCC) 25175 using the disc assay method. To determine AMPs-coding genes, its genome was uploaded to antibiotic and secondary metabolite analysis shell (antiSMASH) 5.0.0 platform and biofilm inhibition was tested using the microtiter plate technique (with a 96-well bottom). Subsequently, the results were assessed using a microplate reader operating at 595 nm wavelength. The isolate was identified as Geobacillus subterraneus, with antibacterial activity against S. mutans, and produced an inhibition zone of 8.40 mm at an optimum pH of 8. The output of AMPs-coding gene showed that AMPs of the isolate were a member of the lanthipeptide class I, or bacteriocin-I group. AMPs of G. subterraneus suppressed the growth of S. mutans biofilm at a supernatant concentration of 5%, with the lowest optical density (OD) value of 0.061 and the highest percentage of biofilm growth inhibition at 28.24%. Based on the results, G. subterraneus derived from the crater of Mount Tangkuban Perahu showed potent antibacterial properties against S. mutans, making it a promising novel S. mutans anti-biofilm candidate.

This study aims to detect Escherichia coli which encodes beta-lactamase Cefotaxime (blaCTX-M) gene from the reproductive tract of Bali cattle with repeat breeder cases. This research was conducted from June to August 2021 using 16 Bali cattle with repeat breeder cases. The reproductive fluids were taken using a plastic sheet gun which was inserted into a Brain Heart Infusion medium, isolated in eosin-methylene blue agar (EMB) and identified using biochemical tests. Antibiotic susceptibility testing of E. coli was carried out using the disc diffusion method. Double-disk approximation test was used to screen the presence of E. coli which produces Extended-spectrum beta-lactamase. The polymerase chain reaction (PCR) method was used to detect the blaCTX-M gene of E. coli and sequences of the blaCTX-M gene were phylogenetically analyzed. The research results obtained three E. coli isolates from 16 reproductive tract fluids of Bali cattle. Antibiotic sensitivity tests showed that 100% of E. coli was resistant to penicillin G and oxytetracycline. 66.66% of E. coli was resistant to cefotaxime (CTX) and gentamicin, and 33.33% of E. coli was resistant to tetracycline. Escherichia coli isolates that were resistant to penicillin and CTX showed positive results in the double-disk approximation test. The results of E. coli detection using PCR showed that three E. coli isolates encoded the blaCTX-M gene located at 370 bp on gel electrophoresis. The results of the phylogenetic analysis showed that E. coli from the reproductive tract of Bali cattle was related to E. coli that encoded blaCTX-M-14 isolated from humans.

Exploring the secondary metabolites with excellent biological activity and pharmacy value from mangrove-derived fungi has attracted great attention. This study aims to isolate endophytic fungi that exhibit antimicrobial activity from Mangrove plants in Siput River Estuaries, Bengkalis Regency, Riau Province, Indonesia. Fifteen isolates of fungal endophytes have been isolated from the root of different Mangrove species, i.e., Aegiceras sp., Lumnitzera racemosa Wild., Avicennia marina (Forssk.) Vierh., Laguncularia racemosa (L) C. F. Gaertn., Sonneratia ovata Backer., Kandelia candel (L) Druce., and Xylocarpus granatum J. Koenig. In vitro, antagonism test was performed for all isolated endophytes against selected pathogenic bacteria. Ten of fifteen endophytes were able to inhibit at least one pathogen with a diameter of inhibition zones ranging from 7,62 ± 0,55 to 18,20 ± 0,98 mm. Three fungi (F15, F2, and F5) with the highest antibacterial activity were subjected to molecular characterization based on the ribosomal region of the internal transcribed spacer by PCR amplification using ITS4 and ITS5 primers. The most potential isolate (F15 from Xylocarpus granatum J. Koenig) showed 99.82% similarity with Botryosphaeria rhodina isolate P130. Its ethyl acetate extract was found positive for terpenoid, phenolic, and flavonoid presence. F2 and F5, originated from L. racemosa Wild. and Avicennia marina (Forssk.) Vierh., had 100% similarity with Fusarium equiseti isolate FUS-34-2 and Aspergillus fumigatus strain DTO 402-H1, respectively. All endophytic fungi are recorded for the first time in Riau Mangrove.

There is an increase rate of hypervirulent Klebsiella pneumoniae (hvKP) that urges the need for preventive and effective immunotherapies, such as vaccine. The YidR gene is a new gene for adherence to host that is conserved in many strain of Klebsiella sp, offering its potency as vaccine candidate. This study aims to isolate the YidRhv gene and analyze the YidRhv protein in silico for future work as vaccine candidate against hypervirulent K. pneumoniae Indonesia strain. Klebsiella pneunomiae is tested for hypervirulency using LAMP-PCR method. The YidRhv gene was amplified via PCR method and the fragment produced was cloned and sequenced. The protein structure and epitope prediction for T-cell and B-cell of YidRhv protein was analyzed using bioinformatics approach. DNA sequence of YidRhv gene strain consisted of 1227 base pairs and showed 99,75 % homology to yidR from classical K pneumoniae. However, several single nucleotide polymorphism were found in this gene. The protein structure demonstrated that the YidRhv is possibly outer membrane protein.

Microalgae have recently been identified as a valuable source of natural bioactive compounds, with potential applications suggested in areas such as food, animal feed, energy production (biofuels), fine chemicals, and pharmaceuticals. However, their chemical diversity remains largely unexplored and it is necessary to determine their potency for further application. This study will explore the diversity of chemical compound from six tropical marine microalgae with untargeted comparative metabolomics approach using a Liquid Chromatography-Orbitrap High Resolution Mass Spectrometry with different mobile phases. Six microalgae species were isolated from Jakarta Bay, Indonesia. Species identification was carried out using morphological and molecular identification. The isolates Chlorella vulgaris InaCC M205 (C5), Chlorella sp. 12 (C12), Tetraselmis subcordiformis InaCC M206 (T2), Tetraselmis sp. 5 (T5), Nannochloropsis oceanica InaCC 207 (N4), and Porphyridium purpureum (P) were then cultivated using f/2 media in 1 L glass photobioreactors, irradiated with daylight lamps (4000 lux) under a 12 h/12 h photoperiod at 25 0C for until cultures reached the stationary phase. Biomass harvesting and measurements were carried out at the end of the culture period. The freeze-dry biomass then was analyzed using liquid chromatography-high resolution mass spectrometry. The results showed that biomass production on day 12 was not significantly different. Biomass production ranged from 0.8 to 1 g/L, with the highest production obtained from the C5 strain and the lowest from the C12 strain. Chromatography analysis revealed that untargeted metabolomics profiling of microalgae, using water-acetonitrile as mobile phase, identified potential compound markers such as N-Methyl-2-pyrrolidone, l-(+)-Cysteine, TBHQ and L-Phenylalanine, which can differentiate between species. Compound such as Cyromazine, Arachidic acid and 13(S)-HOTrE were the potential compound markers when using Water-Methanol as mobile phase. Our research indicates that the metabolomic profile of microalgae can be used to differentiate species depending on the choice of mobile phase.

Objectives
Food products are often contaminated by pathogens and spoilage bacteria. Most of them can form biofilms, a community of cells embedded in protective extracellular matrix layers resistant to harsh conditions, including antibiotics. Therefore, alternative antibiofilm agents are required to overcome biofilm formation. This study aims to determine and quantify the antibiofilm activity of supernatants from plant-associated bacteria against biofilms of foodborne pathogen and food spoilage bacterium, namely Bacillus cereus and Bacillus subtilis.
Results
Plant-associated bacteria (PAB) have shown promising antibiofilm activities against biofilm-forming pathogens in previous studies. Thirteen PAB isolated from Ternate, Indonesia were used in this study. Supernatants of PAB were subjected to antimicrobial activity and quorum quenching detection, both using the well diffusion method. Four supernatants inhibited the growth of B. subtilis, but none affected the growth of B. cereus. Eight supernatants were able to disrupt the quorum sensing system of an indicator bacterium, wild-type Chromobacterium violaceum. Biofilm inhibition and destruction were quantified using 96-well microplates. The highest biofilm inhibition and destruction activities of PAB supernatants against each of B. cereus and B. subtilis biofilms were > 76%, and were later confirmed by light microscope and scanning electron microscope. Brine shrimp lethality assay (BSLA) was conducted and revealed that the selected PAB supernatants were non-toxic. The 16S rRNA gene of PAB were sequenced and they showed similarities to Bacillus, Priestia, and Chryseobacterium. Compounds in the supernatants were determined by GC–MS which revealed contents of fatty acids, ethyl esters, and diketopiperazines. Therefore, PAB supernatants have poten

Objectives
This research aimed to identify and quantify the antibiofilm activity of bioactive compounds from bacteria isolated from rhizosphere and nodule butterfly pea (Clitoria ternatea), rhizosphere clove afo 3 (Syzygium aromaticum), nodule mimosa (Mimosa pudica L.), and soil from gold mining land which were recovered from Ternate, Tidore, Obi Island, and Marotai Island, Eastern part of Indonesia.
Results
Eight supernatants from soil and plant-associated bacteria were found to have quorum quenching activity against Chromobacterium violaceum. All supernatants exhibited antibiofilm activity against biofilm formed by Aeromonas hydrophila and Vibrio harveyi. The supernatant of FT5 showed the highest activity in disrupting (66.59%) and inhibiting (85.63%) the biofilm of A. hydrophila. For V. harveyi, the supernatant of PTM3 showed the highest disruption activity (72.61%), whileRCA7 showed the highest inhibition activity(75.68%). The Gas Chromatography-Mass Spectrometry (GC-MS) identified fatty acids, ester, and diketopiperazine as the compounds related to the antibiofilm activity. Molecular identification revealed that the isolates belong to the genera Bacillus, Priestia, and Chryseobacterium.

The most bothersome weed in rice fields in the Indonesian province of West Java is Monochoria vaginalis (Burm. F.) C. Presl, an aquatic herbaceous plant. Metsulfuron-methyl has long been used in wetland rice in West Java with a high enough intensity. However, the case of Monochoria vaginalis resistance to metsulfuron-methyl herbicides in Indonesia has not been widely reported and investigated. The study aims to (1) classify the resistance level of M. vaginalis toward metsulfuron-methyl, (2) identify Target Site Resistance (TSR) mechanism mutations in the MvALS1 gene of the resistant biotype of M. vaginalis. The Whole Plant Pot Test method was utilized to assess the resistance level of Monochoria vaginalis. Following that, all samples were subjected to DNA sequencing using the PCR method to identify mutations in the MvALS1 gene from the resistant biotype. After then, this study used DUET, a server with an integrated computational methodology, to anticipate the effect of mutations on protein stability. The result showed that Monochoria vaginalis from Rawamerta, Karawang showed a moderate level of resistance to metsulfuron-methyl with a resistance ratio of 6.00, Patokbeusi, Subang showed a low level of resistance to metsulfuron-methyl with a resistance ratio of 3.89, compared to susceptible Monochoria vaginalis. Nucleotide base alignment in the MvALS1 gene revealed that base substitutions occurred in the Monochoria vaginalis biotype from Rawamerta and Patokbeusi, resulting in 5 amino acid substitutions: Ser-64-Ala, Asp-66-Glu, Asn-240-Asp, Glu-426-Asn, and Ser-469-Asn and Sukra: Ser-64-Ala, Asp-66-Glu, and Asn-240-Asp. The analysis showed that S64A, D66E, and N240D stabilize the protein, whereas E426N and S469N destabilize it. This study confirms for the first time that Ser-64-Ala, Asn-240-Asp, and Glu-426-Asn amino acid mutations were found in cases of M. vaginalis resistance to metsulfuron-methyl (ALS inhibitor).

Microalgae offer numerous benefits, including the production of value-added products like omega-3 fatty acids, pigments, and bioactive compounds, as well as their potential use as a third-generation biofuel. Euglena, a unicellular microalgae, is gaining attention as an industrially relevant organism and an emerging cell factory. It exhibits various trophic growth types depending on cultivation conditions. Light intensity and wavelength are critical factors affecting microalgal growth and productivity. This study explores the effects of different light-emitting diode (LED) wavelengths on the growth models and metabolite production of Euglena gracilis from peatland water. The highest specific growth rates were observed under blue and white light in both the logistic and Gompertz models. White light produced the highest biomass yield (0.813 ± 0.261 g.L⁻¹). Our findings indicate that white light is the most effective treatment for enhancing growth and metabolite content, including lipids, carbohydrates, paramylon, and carotenoids. Conversely, green light resulted in the highest protein content. Different illumination treatments led to variations in lipid profiles and fatty acid composition. The highest SFA content was recorded under red light (59.6%), while the highest PUFA content was observed under blue light (31.1%), and the highest MUFA content was detected under red light (18.0%). In conclusion, this study highlights the impact of LEDs on the growth models of E. gracilis and their influence on the production of biomass, carbohydrates, proteins, chlorophyll, carotenoids, paramylon, and fatty acids.

The utilisation of enzymes in the industry has brought numerous benefits and advantages to production processes. Enzymes serve as biocatalysts, efficiently catalyzing reactions and hydrolysis in biochemical processes. However, there are challenges in applying enzymes in the industry, particularly concerning enzyme stability. The obstacle encountered in the production processes involv-ing industrial enzyme applications is the low stability of enzymes when used at high temperatures. Heat-sensitive enzymes undergo damage or denaturation. Thermophilic microorganisms are chosen because they hold the potential to produce thermophilic enzymes. The thermophilic enzymes exhibit better heat stability compared to other enzymes, making them an effective alternative for future industrial production processes. This study aims to isolate thermotoler-ant bacteria from Nglimut Hot Spring sediment, screen for cellulase and amylase-producing isolates, and molecularly identify the best isolate using 16S rRNAbarcode. The results show that 22 bacterial isolates were found in the sediment of a hot spring; TS-14 was the best isolate in producing amylase, with the highest average amylolytic index of 2.38, whereas TS-15 had the highest cellulolytic index of 2.11. Based on 16S rRNA identification, TS-14 showed an homological identity of79% with Bacillus amyloliquefaciens, while TS-15 had a 100% homological identity with Bacillus licheniformis.These re-sults were important as the first step of screening bacterial potential to pro-duce thermophilic enzymes that could be applied in the downstream pro-cessing in future industrial and biotechnology companies.

The presence of ferritin protein in rice plants indicates a regulatory response to environmental iron stress. It prevents the destructive effects of the chain reaction resulting from Fe2+accumulation in the cell. This study aimed to detect and identify the location of the ferritin gene (OsFER) in eight local rice varieties in Indonesia. OsFER2, as the target gene was amplified using PCR, visualized by electrophoresis, and then sequenced. The sequencing results wereanalyzed using DNA Baser, BioEdit, and ClustalX2. The predicted proteins were visualized using the SWISS–MODEL server, Rice Genome Annotation Project Database, and chromosome map tools. The results show that all rice varieties studied have 100% alignment similarity and OsFERcharacteristics on chromosome 11 at LOC_Os11g01530 and chromosome 12 at LOC_Os12g01530, which have striking differences. The complete protein structure of the complex is found on chromosome 12, while only a portion of alpha–helix ferritin is found on chromosome 11. Differences in the position of the ferritin gene at different chromosomes impact the functional changes in the ferritin protein as an iron homeostasis key role. The two genes show different characteristics and are located at different genomic positions, suggesting a potential regulatory impact on the level of iron stress resistance in Indonesian rice varieties.

Advances in fermentation technology, genetic engineering, and enzyme application technology have increased the use of enzymes. Enzymes can be produced by utilizing a source of microorganisms such as bacteria. Proteolytic bacteria or protease enzyme-producing bacteria are found in foods or plants that contain protein such as the brown seaweed Hydroclathrus sp. This study aimed to obtain protease-producing bacterium associated with marine algae Hydroclathrus sp. from waters around Hoga Island of Wakatobi District and identify the organism based on its 16S rRNA gene sequence. Isolation of bacteria from algae samples was carried out with nNutrient aAgar (NA) media, while proteolytic bacteria selection was carried out on sSkim mMilk aAgar (SMA) media. The bacterial isolates producing proteolytic-clear zone on SMA media were then identified targeting the 16S rRNA gene using the PCR (Polymerase Chain Reaction) method with 27F-1492R primers. Based on the isolation results, there were 3 unique colonies of bacteria that could be cultured from algae samples and coded HIHA-1 to HIHA-3 (HIHA stands for Hoga Island Hydrolathrus macroalgae). The selection process for protease-producing bacteria on SMA media resulted in 1 isolate of proteolytic bacteria, namely HIHA-1. Molecular identification by PCR on HIHA-1 isolate resulted in a single DNA band on the electrophoresis gel sized ~1500 bp. The sequencing results showed a DNA sequence with the size of 1421 bp sharing the highest similarity with the bacterium Exiguobacterium aestuarii strain TF-16 (homology level of 99,93%). In conclusion, the proteolytic bacterial isolate HIHA-1 associated with marine brown algae Hydroclathrus sp. was obtained and identified as Exiguobacterium aestuarii strain HIHA-1.

Gaharu bouya oil obtained from distillation of the woods from Gonystylus genus has attracted essential oil industry interest. However, the information about gaharu bouya essential oil profile is limited. The presence of Gonystylus species is also critically endangered on the IUCN Red List. Therefore, exploring the -omics profiles of Gonystylus bancanus, a native plant from Borneo Island, is important for Indonesia to conserve the population. This research investigated the metabolite profiling of G. bancanus oil, especially the volatile components of its essential oils. Distillations were performed in two technical ways: hydrodistillation on a laboratory scale and steam distillation on an industrial scale. According to LC–MS and GC–MS profiles, both essential oils displayed similar chemical compositions. This article also discusses the similarity of the chemical contents of gaharu bouya oil and agarwood oil from the gaharu superior type (Aquilaria) to support the value of the oil. This research also investigated the cytotoxicity of gaharu bouya oil against three cell lines: HeLa, MCF-7, and HT-29.

Basal plate rot disease is an important disease of shallots that causes losses in the field and storage. The disease is caused by Fusariumcomplex species, with different levels of virulence and host susceptibility. The six species of Fusariumspp.reported to cause basal plate rot disease of onion were F. oxysporum f.sp. cepae, F. proliferatum, F. redolens, F. solani, F. acutatumand F. tricinctum. Information about the variation in symptoms and morphology of fungi that cause basal plate rot disease on shallots, especially in Indonesia, is still very limited. Besides, the species that cause basal plate rot disease on shallots has not been molecularly confirmed using specific primers. The objective of this study was to isolate, identify, and study the variationin symptoms of the fungus associated with basal plate rot disease. The fungi were isolated from shallot plants symptomatic of basal plate rot disease collected from Brebes, Central Java Province. Eight isolates of Fusariumwith different morphologies weresuccessfully isolated and identified. Five isolates were identified as F. solani (isolate BC1, BC2, BC3, BBS1, and BBS4), two isolates as F. oxysporum f.sp. cepae (isolate BC4 and BBS6) and one isolate F. proliferatum (isolate BBS5). All eight isolates were pathogenic on shallots with different levels of virulence and symptom variations. BC4 isolate had the highest level of virulence, resulting in plant death, and was identified as F. oxysporum f.sp. cepae.

Liriomyza spp.(Diptera: Agromizidae) is a polyphagous pest that attacks various types of vegetable and ornamental plants throughout the world. The most damaging species are Liriomyza huidobrensis(Blanchard), Liriomyza sativae(Blanchard), and Liriomyza trifolii(Burgess). There have been no reports regarding mapping of population distribution and population genetics of Liriomyzain the Bali region using theCytochrome C Oxidase Subunit I (COI) approach. This research aims to map the population and genetic structure of L. huidobrensis, L. sativaeand L. trifoliion vegetable plants in Bali. This research uses a purposive sampling method which represents vegetable plantings in each district and city in Bali Province. Identification of the Liriomyzaspp.gene structure using the COI approach using LCO and HCO primers. The results of the research found that the spatial distribution of invasive leafminer fly species, namely L. sativaeand L. trifolii, was evenly distributed throughout the city districts in Bali. The distribution pattern of L. trifoliiis uniform with an S2/X value <1, while L. huidobrensisis found in the highland areas with a clustered distribution patternwith an S2/.Thegenetic distance between the three Liriomyzaspecies found in vegetable plants in Bali is much longer. This means that the three Liriomyzaspecies show very high levels of genetic differentiation in mitochondrial and nuclear genes, and differentiation between species in nuclear genes. This genetic variation can influence the ability of insects to quickly adapt to new environments and contribute to population dynamics and dispersal capacity.

Lactococcus garvieae is a gram-positive ovoid cocci bacterium formerly classified as a member of the Lactococcus genus. This study aims to isolate L. garvieae from catfish rearing pond and characterize it as a potential probiotic candidate. L. garvieae was identified and characterized through phenotypic and genotypic observation, genomic % G~C content analysis, cell surface hydrophobicity assays, acidification test, in vitro antagonism, and a profile of antimicrobial activities. The MT597595.1 accession number corresponds to L. garvieae, as determined by a molecular identification test. Biochemical characterization was performed using API 50 CH kit. The genomics %G~C content of L. garvieae was 51.8. Findings from acidification ability tests, in vitro antagonism tests, and the ability of bacteria to grow in broth medium at pH 4 reveal that L. garvieae can inhibit the growth of Aeromonas hydrophila, Streptococcus agalactiae, Streptococcus iniae, and Edwardsiella ictaluri. However, it does not suppress the growth of L. garvieae Edwardsiella tarda. Remarkably, L. garvieae has the ability to reduce the pH of neutral broth medium turning it acidic. Furthermore, L. garvieae’s hydrophobic cell surface exhibited an adhesive, hydrophobic,and protein surface cell content with a compact growth pattern consistent with postive SAT and MATH assay. Antimicrobial activity tests, encompassing 11 antibiotics, disclosed resistance to Nalidixic acid while displaying intermediate sensitivity to Streptomycin and Trimethoprim. In conclusion, L. garvieae demonstrates an inhibitory effect on the growth of pathogenic bacteria, underlining its potential as a probiotic candidate.

Rhizobia are bacteria that symbiosis with host plant, as shown in the root nodules formation, and provide nitrogen that can be absorbed by plants in greater quantities than rhizobacteria. Available Nitrogen, which absorbed by plants, is the essential requirement for plant growth because its role in increasing yield and quality, hence it is needed in greater quantities than other nutrients. The study aimed to determine the macroscopic and microscopic diversity of rhizobia isolates from the groundnut nodules and their potential as PGPRs, andto identify 16S rRNA isolates with the best potential as PGPRs molecularly. The methods used were isolation from root nodules, screening of PGPR potential, molecular identification based on the 16S rRNA gene, and phylogenetic analysis to determine their kinship. Based on the isolation results, 17 Gram–negative isolates were obtained white to pink or orange color on AG media with various colony characteristics in terms of shape, margin, elevation, and texture. KT 20, which was selected as rhizobia isolate with the best potential as PGPR, has ammonium concentration of 23.12 ppm, synthesizes IAAwithaconcentration of 10.36 ppm, and phosphatessolubilization activity, although its ability to synthesize proteases is low. The results of molecular identification of 16S rRNA showed that KT 20 belongs to the Rhizobiumgenus witha similarity of 99.48% andbootstrap valueof 96%.

Green macroalgae, known locally as Latoh, is one of the green seaweeds consumed by the local community in Jepara and is beneficial for health. This study explores the potential of secondary metabolites from seaweed and its symbiotic bacteria as natural food preservatives and antibacterials. Seaweed samples were collected from the seagrass ecosystem of Panjang Island, Jepara, Indonesia. Subsequently, the samples were subjected to scanning electron microscopy analysis, proximate analysis, phytochemical analysis, thin layer chromatography, and high–performance liquid chromatography for amino acid analysis. A sample was subjected to a multistage extraction process using n–hexane, ethyl acetate, and methanol (1:5 w/v), each for 24 h. Symbiotic bacteria from seaweed were isolated, and enzymatic (proteolytic, amylolytic, and cellulolytic) and antibacterial testing against pathogenic bacteria Staphylococcus aureusand Escherichia coliwas conducted using the disc diffusion method. The selected bacteria were subjected to molecular identification. The research showed that Caulerpa lentilliferahad an ash content of 3.24%, protein content of 0.57%, and fat content of 0.337%. Phytochemical analysis shows that the sample contains flavonoids, steroids, and alkaloids. HPLC analysis reveals that Caulerpa lentilliferahas the highest content of aspartic acid (relative area: 11.90%), glutamic acid (relative area: 13.43%), and alanine (relative area: 9.03%). Caulerpa racemosasample shows the highest detector response for glutamic acid (relative area: 12.19%), aspartic acid (relative area: 11.10%), and alanine (relative area: 9.63%). The results indicate that 14 bacterial isolates were successfully isolated, with 6isolates from Caulerpa lentilliferaand 8isolates from Caulerpa racemosa,all exhibiting enzymatic and antibacterial abilities. The research results concluded that the Latoh seaweed species Caulerpa lentillifera andCaulerpa racemosaand their symbiotic bacteria have the potential to be used as food preservatives

Trichoderma has the potential to suppress fungal pathogens and thus control plant diseases, including Phytophthora foot rot, which is the most devastating disease of black pepper in Lampung. Identification of a microorganism can not only rely on its morphological characteristics, but it is also necessary to identify it molecularly at the species level. This research was aimed at identifying the fungus Trichoderma sp. Margodadi isolates at the species level and to know the potential of Trichoderma sp. Margodadi isolates and their secondary metabolites to control P. capsici. This research was conducted from March to November 2021 at the Laboratory of Plant Disease, Department of Plant Protection, and the Laboratory of Agricultural Biotechnology, Faculty of Agriculture, University of Lampung. Identification of Trichoderma was done by morphological characteristics and molecular methods. The ability of Trichoderma to suppress P. capsici was tested by dual culture. The effect of secondary metabolites on the growth of P. capsici was determined in vitro at concentrations of 0% (control), 10%, 20%, 30%, and 40%. The experimental design used was a completely randomized design consisting of five treatments repeated five times. The data obtained from the test were analyzed using ANOVA, followed by the LSD test at 5%. The results of this study showed that Trichoderma sp. Margodadi isolate had a close relationship with Trichoderma asperellum and had the ability as an antagonist to inhibit the growth of P. capsici up to 47.23%, and the secondary metabolites produced could inhibit the growth of P. capsici up to 72.53% with the best concentration of 40%. 

Polyethylene terephthalate (PET) plastic is the most widely used type of plastic that produces waste and causes various environmental and health problems. The treatment of PET plastic waste with chemically and mechanically recycling approaches still has shortcomings, so biological processing using microorganisms or enzymes has new potential. Two bacterial isolates from the Indonesian Culture Collection of NationalResearch and Innovation Agency (InaCC, BRIN), namely isolate InaCC B538 and InaCC B947, were further observed for their potential in PET plastic degradation. Firstly, both isolates were determined by the molecular marker 16S rDNA. The potential of both isolates was measured with following method: 10 days of degradation using PET and PCL substrates, esterase enzyme activity assay, and observation of the PET plastic surface using Scanning Electron Microscope (SEM). Species identification was performed using DNA sequencing of 16S rDNA. InaCC B538 and InaCC B947 were closely related to Bacillus altitudinis TBMAX41 and Alcaligenes faecalis AN-13, respectively. InaCC B947 isolate has a better potential in degrading PET plastic and PCL with a degradation percentage of 0.32% for PET plastic and 3.22% for PCL film for 10 days, respectively, and esterase activity of 0.06 U/ml; while InaCC B538 did not cause weight loss of PET and 2.49% for PCL, respectively, with esterase activity of 0.04 U/ml. The degradation of PET plastic by the isolates InaCC B947 was able to cause damage to the plastic surface leading to the degradation of PET plastic.

The Nias Islands are an archipelago in the western part of North Sumatera, encompassed by the Indian Ocean, and have become a hotspot for demersal and pelagic fishes. These geographical conditions endow the waters around Nias with large fishery resources, leading to their frequent utilization as fishing grounds for Sumatera Island and its surrounding areas. This research aimed to identify the commercially important fish species with the highest catch frequency in the waters of Nias, and this identification marked the initial stage of the sustainable utilization of fish resources. The method used was DNA Barcoding using genes targeting the COI Mitochondrial locus. Therefore, 43samples were collected from several North and South Nias fish landing sites. The obtained species were classified into 3 groups based on the commodity type: demersal fish, pelagic fish, and Choncricthyes. There were 8 species of demersal fish (Caesio caerulaurea, Halichoeres scapularis, Lethrinus ornatus, Mulloidichthys flavolineatus, Parupeneus barberinus, Scarus prasiognathos, Plectropomus leopardus, andVariola albimarginata), displaying genetic distances ranging from 0.002–0.275. Pelagic fish consisted of 5 species, namely Amblygaster clupoides, Caranx ignobilis, Caranx sexfasciatus, Ferdauia ferdau, andScomberomorus commerson, displaying genetic distances ranging from 0.002–0.267. The next commodities were Chondrichthyeswith 9 species (Carcharhinus sealei, Himantura leoparda, Neotrygon kuhlii, Paragaleus randalli, Pateobatis jenkinsii, Rhynchobatuscf laevis, Spyrna lewini, Taeniura meyeni, andUrogymnus granulatus), displaying genetic distance ranging from 0–0.271.This value indicates that migratory species such as Chondrichthyeshave quite extensive movements so that the genetic distance between species and between populations tends to be low.

Pemphis acidulais a wild plant in rocky or sandy coastal areas and mangrove ecosystems. Different geographic characteristics may affect plant adaptability and have an impact on the emergence of various genotypes. This study was performed to reveal the phenetic relationship and genetic variation of P. acidu-lain 3 different areas in Tomini Bay, Gorontalo Province, Indonesia. We took 3 samples from each location and analysed them using 14 morphological char-acters and molecular approaches based on ISSR markers and ITS gene. The results showed that P. acidulaon Olele had bigger sizes in some morphological features compared to the plants in other study areas. The phenetic analysis showed that P. acidulaat Biluhu and Dulanga were more closely related, alt-hough P. acidulaat the 3 locations had 100% similarity. Genetic variation anal-ysis showed the highest genetic similarity based on ISSR markers was found in Dulanga and Biluhu samples (76.8%). Phylogenetic based on ITS gene re-vealed that Olele samples were in the same clade with P. acidula accession from GenBank (genetic distance 0-0.19%), while Biluhu samples were a sister group (genetic distance 24.97-25.03%) even though their percentage identity corre-sponds to P. acidula (81.34%). Plant adaptation to different habitat conditions may affect the genetic diversity of P. acidula.

Background and Aims: Despite the endemicity of Japanese encephalitis virus (JEV) in humans and animals in the Province of Bali, Indonesia, there is little data on whether seroconversion to the virus occurs in pigs, JEV genotypes circulating, and it’s potential mosquito vectors in the area. The aims of this study were to (i) Determine whether JEV infection in Balinese pigs occurs before reaching their sexual maturity, (ii) identify the genotypes of circulating JEV, and (iii) identify potential JEV mosquito vectors at the study sites in urban and peri-urban areas of Bali. Materials and Methods: Sixteen 1-week-old Landrace piglets from two different sows were housed in Denpasar. Similarly, 18 one-week-old mixed-breed piglets of two different sows were housed in Badung Regency. The piglets were bled every 1 to 4 weeks for up to 24 weeks. Serum samples from the 11 piglets were tested for antibodies against JEV, and seroconversion- suspected sera were titrated using an enzyme-linked immunosorbent assay. Blood of seroconverted sera from pigs were tested using polymerase chain reaction (PCR) to detect the genetic sequence of JEV. The mosquitoes in the sentinels were trapped throughout the study period to identify the potential mosquito vectors of JEV. Results: Antibodies were detected in most of the selected piglets’ sera from weeks 1 to 24 of their age. However, sera of pig B9 collected from the sentinel setting in Badung Regency showed a four-fold increase in antibody titer from week 4 to week 8, indicating seroconversion. PCR testing of blood from B9 (pooled blood sample collected from week 5 to week 8) identified JEV nucleic acids, which were phylogenetically classified as belonging to the JEV genotype III. Meanwhile, 1271 of two genera of mosquitoes, Anopheles spp. and Culex spp. were trapped in the pig sentinels. Conclusion: JEV seroconversion likely occurs before the pig reaches sexual maturity in Badung Regency. Sequence data indicate that JEV genotype III is circulating in the pig sentinel setting in the regency; however, circulating genotypes need to be clarified through increased surveillance. Meanwhile, Culex spp. and most likely Culex quinquefasciatus and Anopheles spp. were the dominant mosquitoes present in the study sites set in the urban area of Denpasar and peri-urban areas of Badung, Bali, indicating that these are likely vectors in spread of JEV in the region

This research aims to quantify antiquorum sensing and antibiofilm activity of f phyllosphere bacteria against biofilm formed by pathogenic fish bacteria such as Aeromonas hydrophila, Streptococcus agalactiae, and Vibrio harveyi. Antiquorum sensing assay using Chromobacter violaceum as indicator bacteria and antibiofilm assay showed six phyllosphere bacteria have antiquorum sensing and antibiofilm activities against tested bacteria. The highest inhibition and destruction activity was showed by metabolite of JB 3B and EJB 5 F against A. hydrophila, respectively. Determination using light microscope and scanning electron microscope performed decreaing in biomass of biofilm observed after treated with metabolite from phyllosphere bacteria.