Evidence from the gut microbiota of swarming alates of a vertical transmission of the bacterial symbionts in Nasutitermes arborum (Termitidae, Nasutitermitinae).

Studies on termite symbiosis have revealed that significant symbiont lineages are maintained across generations. However, most studies have focused only on the worker caste. Little is known about the gut microbiota of reproductives, the most probable vectors for transmitting these lineages to offspring. Using 16S rRNA gene-based Illumina MiSeq sequencing, we compared the gut microbiota of swarming alates of the higher termite Nasutitermes arborum with those of their nestmates from the parental colony. The OTU-based alpha diversity indices showed that the gut microbiota of the alates was at least as diverse as those of non-reproductive adults. It was largely dominated by Spirochaetes mostly of the Treponema I cluster (63.1% of reads), the same dominant taxa found in soldiers and workers of this species and in workers of closely related Nasutitermes species. The termite-specific lineages also included other representative taxa such as several clusters of Bacteroidetes and Fibrobacteres-TG3 group. The microbiota of alates was dominated by a core set of host-specific lineages (87% of reads, 77.6% of OTUs), which were always present across all castes/stages. This first comprehensive survey of the microbiota of the founding reproductives of these xylophagous higher termites shows that the bulk of the host endogenous symbionts, mostly taxa that cannot thrive outside the gut, are brought from the parent colony. The royal pair therefore seems to be a key player in the transmission of symbionts across generations and thereby in host-symbiont codiversification. The high proportion of fiber-degrading lineages in their gut suggests a wood-rich diet unlike the larval stages.

Species delimitation and phylogeny in the genus Nasutitermes (Termitidae: Nasutitermitinae) in French Guiana.

Species delimitation and identification can be arduous for taxa whose morphologic characters are easily confused, which can hamper global biodiversity assessments and pest species management. Exploratory methods of species delimitation that use DNA sequence as their primary information source to establish group membership and estimate putative species boundaries are useful approaches, complementary to traditional taxonomy. Termites of the genus Nasutitermes make interesting models for the application of such methods. They are dominant in Neotropical primary forests but also represent major agricultural and structural pests. Despite the prevalence, pivotal ecological role and economical impact of this group, the taxonomy of Nasutitermes species mainly depends on unreliable characters of soldier external morphology. Here, we generated robust species hypotheses for 79 Nasutitermes colonies sampled throughout French Guiana without any a priori knowledge of species affiliation. Sequence analysis of the mitochondrial cytochrome oxidase II gene was coupled with exploratory species-delimitation tools, using the automatic barcode gap discovery method (ABGD) and a generalized mixed Yule-coalescent model (GMYC) to propose primary species hypotheses (PSHs). PSHs were revaluated using phylogenetic analyses of two more loci (mitochondrial 16S rDNA and nuclear internal transcribed spacer 2) leading to 16 retained secondary species hypotheses (RSSH). Seven RSSHs, represented by 44/79 of the sampled colonies, were morphologically affiliated to species recognized as pests in the Neotropics, where they represent a real invasive pest potential in the context of growing ecosystem anthropization. Multigenic phylogenies based on combined alignments (1426-1784 bp) were also reconstructed to identify ancestral ecological niches and major-pest lineages, revealing that Guyanese pest species do not form monophyletic groups.

Mercury species in the nests and bodies of soil-feeding termites, Silvestritermes spp. (Termitidae, Syntermitinae), in French Guiana.

Mercury pollution is currently a major public health concern, given the adverse effects of mercury on wildlife and humans. Soil plays an essential role in speciation of mercury and its global cycling, while being a habitat for a wide range of terrestrial fauna. Soil fauna, primarily soil-feeding taxa that are in intimate contact with soil pollutants are key contributors in the cycling of soil mercury and might provide relevant indications about soil pollution. We studied the enrichment of various mercury species in the nests and bodies of soil-feeding termites Silvestritermes spp. in French Guiana. Soil-feeding termites are the only social insects using soil as both shelter and food and are major decomposers of organic matter in neotropical forests. Nests of S. minutus were depleted in total and mobile mercury compared to nearby soil. In contrast, they were enriched 17 times in methylmercury. The highest concentrations of methylmercury were found in body of both studied termite species, with mean bioconcentration factors of 58 for S. minutus and 179 for S. holmgreni relative to the soil. The assessment of the body distribution of methylmercury in S. minutus showed concentrations of 221 ng g-1 for the guts and even higher for the gut-free carcasses (683 ng g-1), suggesting that methylmercury is not confined to the gut where it was likely produced, but rather stored in various tissues. This enrichment in the most toxic form of Hg in termites may be of concern on termite predators and the higher levels in the food chain that may be endangered through prey-to-predator transfers and bioaccumulation. Soil-feeding termites appear to be promising candidates as bio-indicators of mercury pollution in soils of neotropical rainforest ecosystems.

Variations in the relative abundance of Wolbachia in the gut of Nasutitermes arborum across life stages and castes.

There are multiple forms of interactions between termites and bacteria. In addition to their gut microbiota, which has been intensively studied, termites host intracellular symbionts such as Wolbachia. These distinct symbioses have been so far approached independently and mostly in adult termites. We addressed the dynamics of Wolbachia and the microbiota of the eggs and gut for various life stages and castes of the wood-feeding termite, Nasutitermes arborum, using deep-sequencing of the 16S rRNA gene. Wolbachia was dominant in eggs as expected. Unexpectedly, it persisted in the gut of nearly all stages and castes, indicating a wide somatic distribution in termites. Wolbachia-related sequences clustered into few operational taxonomic units, but these were within the same genotype, acquired maternally. Wolbachia was largely dominant in DNA extracts from the guts of larvae and pre-soldiers (59.1%-99.1% of reads) where gut-resident lineages were less represented and less diverse. The reverse was true for the adult castes. This is the first study reporting the age-dependency of the relative abundance of Wolbachia in the termite gut and its negative correlation with the diversity of the microbiota. The possible mechanisms underlying this negative interaction are discussed.

Profiling the Succession of Bacterial Communities throughout the Life Stages of a Higher Termite Nasutitermes arborum (Termitidae, Nasutitermitinae) Using 16S rRNA Gene Pyrosequencing.

Previous surveys of the gut microbiota of termites have been limited to the worker caste. Termite gut microbiota has been well documented over the last decades and consists mainly of lineages specific to the gut microbiome which are maintained across generations. Despite this intimate relationship, little is known of how symbionts are transmitted to each generation of the host, especially in higher termites where proctodeal feeding has never been reported. The bacterial succession across life stages of the wood-feeding higher termite Nasutitermes arborum was characterized by 16S rRNA gene deep sequencing. The microbial community in the eggs, mainly affiliated to Proteobacteria and Actinobacteria, was markedly different from the communities in the following developmental stages. In the first instar and last instar larvae and worker caste termites, Proteobacteria and Actinobacteria were less abundant than Firmicutes, Bacteroidetes, Spirochaetes, Fibrobacteres and the candidate phylum TG3 from the last instar larvae. Most of the representatives of these phyla (except Firmicutes) were identified as termite-gut specific lineages, although their relative abundances differed. The most salient difference between last instar larvae and worker caste termites was the very high proportion of Spirochaetes, most of which were affiliated to the Treponema Ic, Ia and If subclusters, in workers. The results suggest that termite symbionts are not transmitted from mother to offspring but become established by a gradual process allowing the offspring to have access to the bulk of the microbiota prior to the emergence of workers, and, therefore, presumably through social exchanges with nursing workers.

Increased lead availability and enzyme activities in root-adhering soil of Lantana camara during phytoextraction in the presence of earthworms.

Earthworms are known to increase availability of heavy metals in soils and also play an important role in maintaining the structure and quality of soil. The introduction of earthworms into soils contaminated with metals in the presence of a potential hyperaccumulator has been suggested as an aid for phytoremediation processes. The present study was conducted to evaluate: (i) the effects of earthworms on lead availability in artificially contaminated soil at 500 and 1000 mg kg(-1) Pb in the presence of Lantana camara, a hyperaccumulator, (ii) the effects of earthworms and lead on soil properties such as pH, cation exchange capacity (CEC), organic matter (OM), total and available N, P and K and (iii) soil enzyme activities. Earthworms increased the bioavailable Pb in root-adhering soil by a factor of 2 to 3 in the contaminated soils at concentrations of 500 to 1000 mg Pb kg(-1), respectively. In lead contaminated soils, the presence of earthworms led to a significant decrease in soil pH by about 0.2 but increased CEC by 17% and OM by more than 30%. Earthworm activities also increased the activities of N-acetylglucosamidase, ß-glucosidase, cellulase, xylanase, alkaline and acid phosphatase, urease and fluorescein diacetate assay (FDA). These results indicate that the ecological context for phytoremediation should be broadened by considering plant-soil-earthworm interactions as they influence both plant health and absorption of heavy metals. They also showed that the enzyme activities monitored could serve as useful proxies for phytoremediation capability and, more generally, for soil quality as a whole.

Effect of earthworms on plant Lantana camara Pb-uptake and on bacterial communities in root-adhering soil.

The present study aimed to assess the potential abilities of Lantana camara, an invasive plant species for phytoremediation in the presence of earthworm Pontoscolex corethrurus. Effects of earthworm on growth and lead (Pb) uptake by L. camara plant were studied in soil artificially contaminated at 500 or 1000mg of Pb kg(-1) soil. This species has a promising value for phytoremediation because it can uptake as much as 10% of 1000mgkg(-1) of Pb per year. Moreover, the presence of earthworms enhanced plant biomass by about 1.5-2 times and increased the uptake of lead by about 2-3 times. In the presence of earthworm, L. camara was thus able to uptake up 20% of Pb presence in the soil, corresponding to remediation time of 5 years if all organs are removed. As soil microorganisms are known to mediate many interactions between earthworms and plants, we documented the effect of earthworms on the bacterial community of root-adhering soil of L. camara. Cultivable bacterial biomass of root-adhering soil increased in the presence of earthworms. Similar trend was observed on bacterial metabolic activities. The increase of lead concentrations from 500 to 1000mgkg(-1) did not have any significant effect either on plant growth or on bacterial biomass and global activities but affected the structure and functional diversity of the bacterial community. These results showed that we should broaden the ecological context of phytoremediation by considering plant/microbial community/earthworm interactions that influence the absorption of heavy metals.

Digestive capacities of leaf-cutting ants and the contribution of their fungal cultivar to the degradation of plant material.

Leaf-cutting ants (tribe Attini) are a unique group of ants that cultivate a fungus that serves as a main source of their food. The fungus is grown on fresh leaves that are harvested by workers. We examine the respective contribution of ants and their symbiotic fungus in the degradation of plant material by examining the digestive capacities of seven Attini species in the genera Atta and Acromyrmex. The results show that both, the ants and their mutualistic fungi, have complementary enzymatic activities. Ants are specialized in the degradation of low molecular weight substrates (oligosaccharides and heterosides) whereas the fungus displays high polysaccharidase activity. The two genera Atta and Acromyrmex are not distinguished by a specific enzymatic activity. The seven different mutualistic associations examined display a similar enzymatic profile but have quantitative differences in substrate degradation activities. The respective contribution of ants and the fungus garden in plant degradation are discussed.