Pragmatic Applications and Universality of DNA Barcoding for Substantial Organisms at Species Level: A Review to Explore a Way Forward

DNA barcodes are regarded as hereditary succession codes that serve as a recognition marker to address several queries relating to the identification, classification, community ecology, and evolution of certain functional traits in organisms. The mitochondrial cytochrome c oxidase 1 (CO1) gene as a DNA barcode is highly efficient for discriminating vertebrate and invertebrate animal species. Similarly, different specific markers are used for other organisms, including ribulose bisphosphate carboxylase (rbcL), maturase kinase (matK), transfer RNA-H and photosystem II D1-ApbsArabidopsis thaliana (trnH-psbA), and internal transcribed spacer (ITS) for plant species; 16S ribosomal RNA (16S rRNA), elongation factor Tu gene (Tuf gene), and chaperonin for bacterial strains; and nuclear ITS for fungal strains. Nevertheless, the taxon coverage of reference sequences is far from complete for genus or species-level identification. Applying the next-generation sequencing approach to the parallel acquisition of DNA barcode sequences could greatly expand the potential for library preparation or accurate identification in biodiversity research. Overall, this review articulates on the DNA barcoding technology as applied to different organisms, its universality, applicability, and innovative approach to handling DNA-based species identification.

WEBSITES AS SCIENCE LEARNING ON INVERTEBRATE ANIMALS MATERIALS AT STATE JUNIOR HIGH SCHOOL 1 TEGALDLIMO BANYUWANGI

This study aims to determine the validity of the development of website-based science learning media on invertebrate animal material for grade 7 in Junior High School, and to determine student responses to the development of website-based science learning media on invertebrate animal material. This research was a type of R&D (Research and Development) research which refers to the ADDIE (Analysis, Design, Development, Implementation and Evaluation) development research model of Robert Maribe Branch. The data collection instruments in this study were validation questionnaires and student response questionnaires. Validation was carried out by material experts, media experts, and science teachers. The development trial to determine student responses to the developed product was carried out in two stages, namely a small-scale trial consisting of 6 students and a large-scale trial consisting of 32 students. Based on the results of validation from experts on website-based science learning media, it showed that the assessment of material experts was 96.16%, media expert assessments was 89.12% and assessments from science teachers was 89.83% . In this case, the average percentage of the expert's assessment was 91.70%, which means it was included in the "Very Valid" category and tested on students, and Small-scale trials obtained an average percentage of 94.28%. Then continued large-scale trials and obtained an average percentage of 92.00%. which means “Very Interesting”.

Seasonal variation of genotypes and reproductive plasticity in a facultative clonal freshwater invertebrate animal (Hydra oligactis) living in a temperate lake

Facultative sexual organisms combine sexual and asexual reproduction within a single life cycle, often switching between reproductive modes depending on environmental conditions. These organisms frequently inhabit variable seasonal environments, where favourable periods alternate with unfavourable periods, generating temporally varying selection pressures that strongly influence life history decisions and hence population dynamics. Due to the rapidly accelerating changes in our global environment today, understanding the dynamics of and genetic changes in facultative sexual populations inhabiting seasonal environments is critical to assess and prepare for additional challenges that will affect such ecosystems. In this study we aimed at obtaining insights of the seasonal population dynamics of the facultative sexual freshwater cnidarian Hydra oligactis through a combination of Restriction-site Associated Sequencing (RAD-Seq) genotyping and the collection of phenotypic data on the reproductive strategy of field-collected hydra strains. We found no significant genetic change during the two years in the study population. Clone lines were detected between seasons and even years, suggesting that clonal lineages can persist for a long time in a natural population. We also found that distinct genotypes differ in sexual reproduction frequency, but these differences did not affect whether genotypes reappeared across samplings. Our study describes changes in population genetic structure across the seasons in a hydra population for the first time, providing key insights into the biology of the species, while also contributing to understanding the population biology of facultative sexual species inhabiting freshwater ecosystems.

INVERTEBRATE SPECIES COMPOSITION (Invertebrata) PESTS OF AGROCENOSES OF WHITE CLOVER (Trifolium repens L.) AND THE RELEVANCE OF PROTECTIVE MEASURES

The area of natural growth and the scale of economic use of white clover among perennial legumes are spatially one of the most global. Due to the constant presence of wild white clover in many natural phyto-cenoses of most agricultural landscapes of different geographical zones, a certain complex of phytophages has evolved from various classes of invertebrate animal organisms that feed on various parts of this plant and reproduce on it. With the introduction of white clover into the culture and the spread of production crops of this plant over large areas, more favorable conditions are created for uncontrolled mass re-production of pests, which can cause already economically and economically significant crop losses. White clover is affected by multi-eating and specialized pests, the damage from which is determined by their biological characteristics and climatic conditions. The high population of white clover crops with a complex of herbivorous invertebrates implies constant monitoring and control of the species composition of the harmful fauna of white clover crops in order to organize, if necessary, protective measures against them. This issue is especially relevant for seed crops. The main pests of seed stands are considered to be weevils of the genera Apion Herbs., Phytonomus Herbs., Sitona Germar., Hypera Germar., which can re-duce seed yield by 50% or more. To reduce the economic losses of the crop on white clover, an integrated protection system should be implemented using chemical, biological and agrotechnical pest control meas-ures based on taking into account their economic harmfulness thresholds. One of the important directions of increasing the efficiency of the production use of white clover is the development of varieties of this crop that are resistant to damage by pathogenic organisms and pests.

Newly Isolated Animal Pathogen Corynebacterium silvaticum Is Cytotoxic to Human Epithelial Cells

Corynebacterium silvaticum is a newly identified animal pathogen of forest animals such as roe deer and wild boars. The species is closely related to the emerging human pathogen Corynebacterium ulcerans and the widely distributed animal pathogen Corynebacterium pseudotuberculosis. In this study, Corynebacterium silvaticum strain W25 was characterized with respect to its interaction with human cell lines. Microscopy, measurement of transepithelial electric resistance and cytotoxicity assays revealed detrimental effects of C. silvaticum to different human epithelial cell lines and to an invertebrate animal model, Galleria mellonella larvae, comparable to diphtheria toxin-secreting C. ulcerans. Furthermore, the results obtained may indicate a considerable zoonotic potential of this newly identified species.

KEANEKARAGAMAN BINTANG LAUT (Asteroidea) DI PANTAI LAMALAKA KECAMATAN ILE BOLENG KABUPATEN FLORES TIMUR

Sea star is an invertebrate animal that is included in the phylum Echinoderms from the Asteroidea class. Radial symmetry is shaped and generally has five or more arms and does not have a frame that helps movement. This study aims to determine the diversity of sea star (Asteroidea) in Lamalaka Beach, Ile Boleng District, East Flores Regency.This research will be conducted in July-August 2019, the location of the study is on the Lamalaka coast, Ile Boleng District, East Flores Regency.The method used is a survey method, based on the presence of Asteroidea species that are considered to represent the area. The steps in data collection are field survey, observation, identifying species in each plot, observing and recording their marphological characteristics.Sampling uses the technique of determining the location of sampling using Area sampling. To identify based on Human and Deloach identification guidelines, 2010. Analysis of the data used in this research is quantitative descriptive and the composition of Asteroidea species can be stated based on visual observations calculating species density, relative density, and dominance index, diversity.Based on the results of the study, found 4 species of Asteroidea, namely Protoreaster nodulosus, Linckia laevigata, Echinaster luzonicus, Astropecten polyacanthus. The highest species density calculation results were found in P. nodulosus species with a value of 0.78 ind / m2. while the lowest species density was found in L.laevigata species with a value of 0.63 ind / m2. The highest relative density was found in P. nodulosus species with a value of 27.64%, and the lowest relative density found in L.laevigata species with a value of 22.35 %. The index of starfish species dominance in Lamalaka Beach, obtained a value of D <0.5, indicates that there are no species that dominate in Lamalaka Beach. The existence of starfish in Lamalaka Beach, Ile Boleng District, East Flores Regency needs to be preserved. One form of effort to preserve it is to conduct monitoring with routine monitoring to monitor the condition of the sea star population in these waters. Besides that, it is also necessary to increase the utilization of sea stars as one of the natural tourism objects and as an Ecoedutourism tourism object. Thus, the presence of starfish will be felt by the wider community so that awareness to maintain the existence of starfish will be increased.

Expression Pattern of Nitric Oxide Synthase during Development of the Marine Gastropod Mollusc, Crepidula fornicata

Nitric Oxide (NO) plays a key role in the induction of larval metamorphosis in several invertebrate phyla. The inhibition of the NO synthase in Crepidula fornicata, a molluscan model for evolutionary, developmental, and ecological research, has been demonstrated to block the initiation of metamorphosis highlighting that endogenous NO is crucial in the control of this developmental and morphological process. Nitric Oxide Synthase contributes to the development of shell gland, digestive gland and kidney, being expressed in cells that presumably correspond to FMRF-amide, serotoninergic and catecolaminergic neurons. Here we identified a single Nos gene in embryonic and larval transcriptomes of C. fornicata and studied its localization during development, through whole-mount in situ hybridization, in order to compare its expression pattern with that of other marine invertebrate animal models.

Another Example of Conditioned Taste Aversion: Case of Snails

Conditioned taste aversion (CTA) in mammals has several specific characteristics: (1) emergence of a negative symptom in subjects due to selective association with a taste-related stimulus, (2) robust long-term memory that is resistant to extinction induced by repeated presentation of the conditioned stimulus (CS), (3) a very-long-delay presentation of the unconditioned stimulus (US), and (4) single-trial learning. The pond snail, Lymnaea stagnalis, can also form a CTA. Although the negative symptoms, like nausea, in humans cannot be easily observed in invertebrate animal models of CTA, all the other characteristics of CTA seem to be present in snails. Selective associability was confirmed using a sweet sucrose solution and a bitter KCl solution. Once snails form a CTA, repeated presentation of the CS does not extinguish the CTA. A long interstimulus interval between the CS and US, like in trace conditioning, still results in the formation of a CTA in snails. Lastly, even single-trial learning has been demonstrated with a certain probability. In the present review, we compare, in detail, CTA in mammals and snails, and discuss the possible molecular events in CTA.