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Pollinating insects are of vital importance for the ecosystem and their drastic decline imposes severe consequences for the environment and humankind. The comprehension of their interaction networks is the first step in order to preserve these highly complex systems. For that purpose, the following study describes a protocol for the investigation of honey bee pollen samples from different agro-environmental areas by DNA extraction, PCR amplification and nanopore sequencing of the barcode regions rbcL and ITS. It was shown, that the most abundant species were classified consistently by both DNA barcodes, while species richness was enhanced by single-barcode detection of less abundant species. The analysis of the the different landscape variables exhibited a decline of species richness, Shannon diversity index, and species evenness with increasing organic crop area. However, sampling was only carried out in August and further investigations are suggested to display a more complete picture of honey bee foraging throughout the seasons.
In this work, a protocol for portable nanopore sequencing of DNA from pollen collected from honey bees, bumble bees, and wild bees was developed. DNA metabarcoding is applied to identify genera within the mixed DNA samples. The DNA extraction and ITS and ITS2 PCR parameters tested for this purpose were applied to the collected pollen sample and the amplicons were then decoded using the Flongle sequencer adapter from Oxford Nanopore Technologies. It is shown that the main pollinator resources at the different sites can be identified in percentage proportions. The protocol generated in this study can be used for further ecological questions.
Assessment of COI and 16S for insect species identification ti determine the diet of city bats
(2023)
Despite the numerous benefits of urbanization to human living conditions, urbanization has also negatively affected humans, their environment, and other organisms that share urban habitats with humans. Undoubtedly adverse while some wild animals avoid living in urban areas, others are more tolerant or prefer life in urban habitats. There are more than 1,400 species of bats in the world.
Therefore, they have the potential to contribute significantly to the mammalian biodiversity in urban areas. Insectivorous bats species play a key role in agriculture by improving yields and reducing chemical pesticide costs. Using metabarcoding, it is possible to determine the prey consumed by these noctule mammals based on the DNA fragments in their fecal pellets. This study
aimed to evaluate COI and 16S metabarcodes for insect species identification to determine the diet of metropolitan bats. For this purpose, COI and 16S metabarcodes were extracted, amplified, and sequenced from 65 bat feces collected in the Berlin metropolitan areas. Following a taxonomic annotation, I found that 73% of all identified insects could only be detected using the COI method, while 15% could be recovered using the 16S approach. Just 12% of all detected insects were identified simultaneously by both markers. According to this result, COI is more suitable for the taxonomic identification of insects from bat feces. However, given the bias of COI primers, it is recommended to use both markers for a more precise estimation of species diversity. Additionally,based on the insect species identified, I noticed that urban bats fed mainly on Diptera, Coleoptera,and Lepidoptera. The bat species Nyctalus noctula was most abundant in the samples. His diet analysis revealed that 91% of the samples contained the insect species Chironomus plumosus. 14 pest insect species were also found in his diet.