Metabarcoding, the coupling of DNA-based species identification and high-throughput sequencing, offers enormous promise for arthropod biodiversity studies but factors such as cost, speed and ease-of-use of bioinformatic pipelines, crucial for making the leapt from demonstration studies to a real-world application, have not yet been adequately addressed. Here, four published and one newly designed primer sets were tested across a diverse set of 80 arthropod species, representing 11 orders, to establish optimal protocols for Illumina-based metabarcoding of tropical Malaise trap samples. Two primer sets which showed the highest amplification success with individual specimen polymerase chain reaction (PCR, 98%) were used for bulk PCR and Illumina MiSeq sequencing. The sequencing outputs were subjected to both manual and simple metagenomics quality control and filtering pipelines. We obtained acceptable detection rates after bulk PCR and high-throughput sequencing (80-90% of input species) but analyses were complicated by putative heteroplasmic sequences and contamination. The manual pipeline produced similar or better outputs to the simple metagenomics pipeline (1.4 compared with 0.5 expected:unexpected Operational Taxonomic Units). Our study suggests that metabarcoding is slowly becoming as cheap, fast and easy as conventional DNA barcoding, and that Malaise trap metabarcoding may soon fulfill its potential, providing a thermometer for biodiversity.
Seaweeds have been used as a food for centuries in Asia and are increasingly exploited as a source for dietary supplements, animal feed, chemicals, and biofuels. In recent years, there has been an increase in the prevalence of diseases and pests in these aquaculture crops, with a subsequent reduction in their quantity and commercial value. In this article, we review diseases that have been reported in the scientific literature for species of red and brown seaweeds. We have focused on the major seaweed crops grown in Asia, where much of this production is centered. We also provide information on disease management and biosecurity and some observations on future directions.
Indonesia is the world largest producer of the red seaweeds Kappaphycus and Eucheuma; however, this country is facing significant challenges such as disease outbreaks, epiphyte infestations and a loss in seedling quality. Biosecurity practices have been widely adopted in other aquaculture sectors and when enforced can help to limit the introduction and spread of diseases and pests. To assess current capacity for biosecurity in seaweed aquaculture in Indonesia, a systematic analysis of policy frameworks including legislation, regulatory tools, and national standards was conducted. Biosecurity themes and risks were used to evaluate current national biosecurity content. The results identified major challenges faced by the industry in order to implement biosecurity policies in practice. Barriers to implementation included unspecific reference to the seaweed aquaculture sector, limited variety of approaches to biosecurity, limited use of up-to-date scientific evidence, insufficient guidance for the use of precaution and insufficient inclusion of specific biosecurity hazards. In general, although national regulations are currently under revision, current policies indicate a lack of clarity where biosecurity is included. Six recommendations are suggested to incorporate proactive biosecurity actions into current frameworks, with the aim of improving the health and sustainability of the seaweed aquaculture sector in Indonesia.
Bactrocera latifrons is a serious pest of solanaceous fruits and Bactrocera umbrosa is a pest of Artocarpus fruits, while Bactrocera melastomatos infests the fruit of Melastomataceae. They are members of the subgenus Bactrocera. We report here the complete mitochondrial genome of these fruit flies determined by next-generation sequencing and their phylogeny with other taxa of the subgenus Bactrocera. The whole mitogenomes of these three species possessed 37 genes namely, 13 protein-coding genes (PCGs), 2 rRNA and 22 tRNA genes. The mitogenome of B. latifrons (15,977 bp) was longer than those of B. melastomatos (15,954 bp) and B. umbrosa (15,898 bp). This difference can be attributed to the size of the intergenic spacers (283 bp in B. latifrons, 261 bp in B. melastomatos, and 211 bp in B. umbrosa). Most of the PCGs in the three species have an identical start codon, except for atp8 (adenosine triphosphate synthase protein 8), which had an ATG instead of GTG in B. umbrosa, whilst the nad3 (NADH dehydrogenase subunit 3) and nad6 (NADH dehydrogenase subunit 6) genes were characterized by an ATC instead of ATT in B. melastomatos. The three species had identical stop codon for the respective PCGs. In B. latifrons and B. melastomatos, the TΨC (thymidine-pseudouridine-cytidine)-loop was absent in trnF (phenylalanine) and DHU (dihydrouracil)-loop was absent in trnS1 (serine S1). In B. umbrosa, trnN (asparagine), trnC (cysteine) and trnF lacked the TψC-loop, while trnS1 lacked the DHU-stem. Molecular phylogeny based on 13 PCGs was in general concordant with 15 mitochondrial genes (13 PCGs and 2 rRNA genes), with B. latifrons and B. umbrosa forming a sister group basal to the other species of the subgenus Bactrocera which was monophyletic. The whole mitogenomes will serve as a useful dataset for studying the genetics, systematics and phylogenetic relationships of the many species of Bactrocera genus in particular, and tephritid fruit flies in general.
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