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Given the social sensitivity in the area, the process of evaluating the EIA has followed the highest standards in order to protect the population in isolation and initial contact, which is why it has requested the company to carry out the exploration in a sequential manner (the different activities can't be done simultaneously), medical control of its workers, and limits on people entering its concession, among other things. At the same time, the "no contact" principle will be respected, which is why, among other measures, it has been requested that the area of exploration in the north-east of the concession is limited in order not to affect the possible transit of people in isolation in the River Paquiria basin.
When the plant has outgrown its container, root pruning is advisable. Pull roots away from the root mass then cut them back to within 1 inch of the soil mass. An alternative method is to make three or four vertical cuts 1 inch deep in the soil ball on the opposite sides of the root ball.
Through a broader phylogenetic analysis of these Camponotus AAB and other AAB lineages, we present the first evidence of a novel, monophyletic, and deeply divergent clade of ant-associated gut microbiota positioned in the Acetobacteraceae. We found that the 16S rDNA gene of these ant-associated AAB shows accelerated nucleotide substitution rates, elevated AT content, and decreased predicted stability of the rRNA secondary structure. These features resemble patterns found in coevolved, socially-transmitted gut associates of bumblebees [28]. Such patterns have not been found in other well studied ant associates, such as Opitutales taxa often associated with Cephalotes hosts [22, 29, 30], as we demonstrate here. These results suggest that AAB may form a persistent association with ants, with significant impacts on the evolutionary trajectory of these bacteria.
Phylogenetic analyses of 16S rDNA sequences showed that AAB1 and AAB2 group together, along with other Acetobacteraceae OTUs sampled from ant hosts or ant nests. This group of ant-associated Acetobacteraceae form a deeply divergent, monophyletic clade that is well supported based on Bayesian analysis (1.0 posterior probability; Fig. 1) and maximum likelihood analysis (100 % bootstrap support, Additional file 5).
Several lines of evidence support the hypothesis that Acetobacteraceae is a frequent gut associate in Camponotus species and perhaps other ant groups. First, 16S rDNA amplicon sequencing of local C. chromaiodes microbiota showed that at least one of two Acetobacteraceae OTUs (AAB1 or AAB2) occurred in most host samples analyzed and was present in both workers and mated queens. Second, although some of our samples were based on whole ant gasters, we confirmed through analysis of dissected samples that AAB1 and AAB2 indeed occur in the gut tract of C. chromaiodes. Third, using AAB-specific PCR primers, we detected AAB1 in additional Camponotus species and regions: C. castaneus from North Carolina, and C. castaneus and C. pennsylvanicus from Massachusetts. Fourth, our phylogenetic analysis documented that AAB1 and AAB2 belong to a deeply diverging, monophyletic group that includes associates of diverse ant species (detailed below) and, based on current data, is restricted to ant associates (Fig. 1; Clade B in Fig. 2). This phylogenetic pattern suggests that the clade may be specialized for association with ants. Fifth, at the 16S rDNA gene, molecular evolution of the ant-associated AAB clade exhibit trends that resemble stably inherited symbionts: significantly accelerated rates of evolution, AT-biased sequence composition, and (perhaps a result of AT bias) destabilization of predicted rRNA stability. Combined, these data suggest the association of this AAB clade with ants may be specialized and quite old.
Exploration of transmission routes and caste variation will benefit from a deeper sampling of the gut community. While our efforts to deplete the abundance of 16S rDNA amplicons from Blochmannia were successful, Wolbachia contributed significantly to our sequencing effort. In the context of gut community surveys, the alimentary tract is relatively large among the Formicidae, and likely amenable to microdissection of luminal contents for targeted characterization of those communities, as has been performed in termites a [2]. Combining microdissection with high-throughput sequencing [60] will facilitate deeper exploration of gut communities among insects that also house abundant endosymbionts near the gut.
In concert, the data presented here suggest that Acetobacteraceae symbionts represent abundant and dominant members of the gut community of Camponotus chromaiodes. The two most abundant members, AAB1 and AAB2, belong to a novel and deeply divergent, monophyletic clade of ant-associated bacteria. Relatives of these taxa were recovered from two additional Camponotus species, and are known from prior studies to occur in additional ant groups. Members of this ant-specific AAB clade display patterns of molecular evolution at the 16S rDNA gene that resemble patterns found in socially transmitted gut symbionts of bees and, in a more extreme form, cospeciating, maternally transmitted gut associates of stinkbugs. Specifically, these features include elevated AT content, a ~7.6-fold increase in substitution rates at 16S rDNA, and a notable decrease in the predicted stability of 16S rRNA. Cumulatively, these results are consistent with the hypothesis that this AAB clade may represent a long-term gut associate of ants.
Atma is a sonic gateway into new dimensions of sounds that have been left unexplored until now. Our interactive animated performance interface has been carefully crafted to enhance the exploration of these new sonic realms. Featuring a crystalline ocean and an emerging ephemeral being, opening a gateway into these new dimensions. 2b1af7f3a8
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