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Reflective architecture The study will support a scale-up study to optimize the results of the microbiome surveys in order to provide important insights into large scale management. We will also look into how to manage the size of the microbiome of livestock and fishes as well as the relationship with their health and welfare. Methods Microbiome sampling Livestock Roots of the plants will be collected from animals present in large herds. The crop or other part of the animal will be sampled. Prokaryote Fresh feces will be collected directly from cattle and pigs. Eukaryote When possible, depending on the species, the crop will be sampled. Fishes The water where the fishes live will be sampled and used for the collection of the animal feces. Fresh feces will be collected directly from fish. Protein 3D mesh DNA extraction DNA will be extracted using the DNeasy blood and tissue kit (Qiagen), following manufacturer’s recommendations. 16S rDNA amplicon In order to establish the most representative markers for prokaryotes and eukaryotes, we will amplify the 16S rRNA gene of prokaryotes and the 18S rRNA gene of eukaryotes. RNA extraction RNA will be extracted from the frozen samples using the E.Z.N.A.® Fungal/Bacterial RNA Mini Kit (Omega Bio-Tek, Inc., Norcross, GA, USA). cDNA synthesis All the extracted RNA will be transcribed into cDNA using the Superscript III kit (Invitrogen, Thermo Fisher Scientific, Inc., Waltham, MA, USA). The four steps of a typical PCR reaction are: PCR reactions will be performed on the cDNA using the following primers;Q: Clicking
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