Recent studies describe the use of UAVs in collecting blow samples from large whales to analyze the microbial and viral community in exhaled air. Unfortunately, attempts to collect blow from small cetaceans have not been successful due to their swimming and diving behavior. In order to overcome these limitations, in this study we investigated the application of a specific sampling tool attached to a UAV to analyze the blow from small cetaceans and their respiratory microbiome. Preliminary trials to set up the sampling tool were conducted on a group of 6 bottlenose dolphins (Tursiops truncatus) under human care, housed at Acquario di Genova, with approximately 1 meter distance between the blowing animal and the tool to obtain suitable samples. The same sampling kit, suspended via a 2 meter rope assembled on a waterproof UAV, flying 3 meters above the animals, was used to sample the blows of 5 wild bottlenose dolphins in the Gulf of Ambracia (Greece) and a sperm whale (Physeter macrocephalus) in the southern Tyrrhenian Sea (Italy), to investigate whether this experimental assembly also works for large whale sampling. In order to distinguish between blow-associated microbes and seawater microbes, we pooled 5 seawater samples from the same area where blow samples’ collection were carried out. The the respiratory microbiota was assessed by using the V3-V4 region of the 16S rRNA gene via Illumina Amplicon Sequencing. The pooled water samples contained more bacterial taxa than the blow samples of both wild animals and the sequenced dolphin maintained under human care. The composition of the bacterial community differed between the water samples and between the blow samples of wild cetaceans and that under human care, but these differences may have been mediated by different microbial communities between seawater and aquarium water. The sperm whale’s respiratory microbiome was more similar to the results obtained from wild bottlenose dolphins. Although the number of samples used in this study was limited and sampling and analyses were impaired by several limitations, the results are rather encouraging, as shown by the evident microbial differences between seawater and blow samples, confirmed also by the meta-analysis carried out comparing our results with those obtained in previous studies. Collecting exhaled air from small cetaceans using drones is a challenging process, both logistically and technically. The success in obtaining samples from small cetacean blow in this study in comparison to previous studies is likely due to the distance the sampling kit is suspended from the drone, which reduced the likelihood that the turbulence of the drone propeller interfered with successfully sampling blow, suggested as a factor leading to poor success in previous studies.

最近的研究描述了无人机在从大鲸鱼收集打击样本中的用途，以分析呼出空气中的微生物和病毒群落。不幸的是，由于它们的游泳和潜水行为，试图从小鲸类身上收集打击的尝试并未成功。为了克服这些局限性，在这项研究中，我们研究了连接到无人机的特定采样工具的应用，以分析来自小鲸类动物及其呼吸微生物组的打击。对一组6只宽吻海豚（Tursiops truncatus）在人类护理下，安置在热那亚（Acquario di Genova）上，吹制动物与工具之间的距离约为1米，以获取合适的样品。使用相同的采样套件，该套件通过一条2米长的绳索悬挂在防水无人机上，并悬挂在动物上方3米处，用于在Ambracia湾（希腊）和抹香鲸（Physeter macrocephalus）的打击下，对5只野生宽吻海豚的打击进行采样。）在第勒尼安海南部（意大利）进行调查，以调查该实验装置是否也适用于大型鲸鱼采样。为了区分与打击相关的微生物和海水微生物，我们从进行打击样品收集的同一地区收集了5个海水样品。通过Illumina扩增子测序，使用16S rRNA基因的V3-V4区域评估呼吸道菌群。合并的水样比野生动物和人类照管的测序海豚的打击样中细菌类群更多。在水样本之间以及野生鲸类的打击样本之间和在人类的照料下，细菌群落的组成有所不同，但是这些差异可能是由海水和水族馆水之间的不同微生物群落介导的。抹香鲸的呼吸微生物组与野生宽吻海豚获得的结果更为相似。尽管本研究中使用的样品数量有限，并且采样和分析受到若干限制的影响，但结果却令人鼓舞，如海水和吹气样品之间明显的微生物差异所表明，并通过比较进行的荟萃分析证实我们的结果与以前的研究结果一致。从后勤和技术上来说，使用无人机从小型鲸类中收集呼出的空气是一个具有挑战性的过程。与先前的研究相比，本研究中从鲸类小型打击中成功获取样本的原因可能是采样套件与无人机的悬挂距离，