Leeches are found in terrestrial, aquatic, and marine habitats on all continents. Sanguivorous leeches have been used in medicine for millennia. Modern scientific uses include studies of neurons, anticoagulants, and gut microbial symbioses. Hirudo verbana, the European medicinal leech, maintains a gut community dominated by two bacterial symbionts, Aeromonas veronii and Mucinivorans hirudinis, which sometimes account for as much as 97% of the total crop microbiota. The highly simplified gut anatomy and microbiome of H. verbana make it an excellent model organism for studying gut microbial dynamics. The North American medicinal leech, Macrobdella decora, is a hirudinid leech native to Canada and the northern U.S.A. In this study we show that M. decora symbiont communities are very similar to those in H. verbana. We performed an extensive study using field-caught M. decora and purchased H. verbana from two suppliers. Deep sequencing of the V4 region of the 16S rRNA gene allowed us to determine the core microbiome of M. decora as consisting of Bacteroides, Aeromonas, Proteocatella, and Butyricicoccus. The analysis revealed that the composition of the gut microbiome of the two leech species was significantly different at all taxonomic levels. The R2 value was highest at the genus and ASV levels and much lower at phylum, class, and order taxonomic levels. The gut and bladder microbial communities were distinct. We propose that M. decora is an alternative to H. verbana for studies of wild-caught animals and provide evidence for the conservation of digestive-tract and bladder symbionts in annelid models. IMPORTANCE Building evidence implicates the gut microbiome in critical animal functions such as regulating digestion, nutrition, immune regulation, and development. Simplified, phylogenetically diverse models for hypotheses testing are necessary because of the difficulty of assigning causative relationships in complex gut microbiomes. Previous research used Hirudo verbana as a tractable animal model of digestive-tract symbioses. Our data show that Macrobdella decora may work just as well without the drawback of being an endangered organism and with the added advantage of easy access to field-caught specimens. The similarity of the microbial community structure of species from two different continents reveals the highly-conserved nature of the microbial symbionts in sanguivorous leeches.
24Leeches are found in terrestrial, aquatic, and marine habitats on all continents. Sanguivorous 25 leeches have been used in medicine for millennia. Modern scientific uses include studies of 26 neurons, anticoagulants, and gut microbial symbioses. Hirudo verbana, the European medicinal 27 leech, maintains a gut community dominated by two bacterial symbionts, Aeromonas veronii and 28Mucinivorans hirudinis, which sometimes account for as much as 97% of the total crop 29 microbiota. The highly simplified gut anatomy and microbiome of H. verbana make it an 30 excellent model organism for studying gut microbial dynamics. The North American medicinal 31 leech, Macrobdella decora, is a hirudinid leech native to Canada and the northern U.S.A. In this 32 study we show that M. decora symbiont communities are very similar to those in H. verbana. 33 This similarity allowed for an extensive study in which wild caught animals were sampled to 34 determine effects of geographic separation, time of collection, and feeding on the microbiome. 35Through 16S V4 rRNA deep sequencing we show that: i) the M. decora gut and bladder 36 microbial communities are distinct, ii) the M. decora gut community is affected by feeding and 37 long periods of starvation, and iii) geographic separation does not appear to affect the overall gut 38 microbial community structure. We propose that M. decora is a replacement for H. verbana for 39 studies of wild-caught animals and offer evidence for the conservation of annelid symbionts. 40Successful culturing and comparison of dominant symbionts from M. decora and H. verbana 41 will provide the ability to assess host-symbiont co-evolution in future work. 42 43 44 45 IMPORTANCE 46 48 difficulty of assigning causative relationships in complex gut microbiomes a simplified model 49 for testing hypotheses is necessary. Previous research in Hirudo verbana has suggested this 50 animal as a highly simplified and tractable animal model of gut symbioses. Our data show that 51Macrobdella decora may work just as well as H. verbana without the drawback of being an 52 endangered organism and with the added convenience of easy access to field-caught specimens. 53The similarity of the microbial community structure of species from two different continents 54 reveals the highly-conserved nature of the microbial symbionts in sanguivorous leeches and 55 confirms the medicinal leech as a highly simplified, natural animal model in which to study gut 56 symbioses. 57 58 59 4), resulting in the name medicinal leech, Hirudo medicinalis Linnaeus, 1758. As our 64 understanding of hirundinid taxonomy improved, Hirudo medicinalis was subdivided into 65 additional species including H.verbana Carena 1820 and H.orientalis Utevsky & Trontelj, 2005 66 (5, 6). Since 2004 in the United States, only H. medicinalis and H. verbana are approved for use 67as a medical device and must be shipped from suppliers in Europe (5). Although it shares the 68 same common name, the North American medicinal leech, Macrobdella decora, was rarely used 69 ...
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