Complexity and Variability of Gut Commensal Microbiota in Polyphagous Lepidopteran Larvae

Tang, Xiaoshu; Freitak, Dalial; Vogel, Heiko; Ping, Liyan; Shao, Yongqi; Cordero, Erika Arias; Andersen, Gary L.; Westermann, Martin; Heckel, David G.; Boland, Wilhelm

doi:10.1371/journal.pone.0036978

Cited by 172 publications

(181 citation statements)

References 43 publications

(68 reference statements)

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“…This bacterium had 93% sequence similarity to an unclassified bacterium isolated from the gut of Apis mellifera [33] and 84% sequence similarity to an unclassified Clostridium species isolated from the gut of the moth Spodoptera littoralis [34]. This unclassified Firmicute was proportionally abundant in nearly all antifungal and A. aggregata-treated samples and in seven of the no-treatment control samples, Figure 4.…”

Section: (B) Fungal Community Diversitymentioning

confidence: 95%

Interactions between fungi and bacteria influence microbial community structure in the Megachile rotundata larval gut

2014

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Recent declines in bee populations coupled with advances in DNAsequencing technology have sparked a renaissance in studies of bee-associated microbes. Megachile rotundata is an important field crop pollinator, but is stricken by chalkbrood, a disease caused by the fungus Ascosphaera aggregata. To test the hypothesis that some gut microbes directly or indirectly affect the growth of others, we applied four treatments to the pollen provisions of M. rotundata eggs and young larvae: antibacterials, antifungals, A. aggregata spores and a no-treatment control. We allowed the larvae to develop, and then used 454 pyrosequencing and quantitative PCR (for A. aggregata) to investigate fungal and bacterial communities in the larval gut. Antifungals lowered A. aggregata abundance but increased the diversity of surviving fungi. This suggests that A. aggregata inhibits the growth of other fungi in the gut through chemical or competitive interaction. Bacterial richness decreased under the antifungal treatment, suggesting that changes in the fungal community caused changes in the bacterial community. We found no evidence that bacteria affect fungal communities. Lactobacillus kunkeei clade bacteria were common members of the larval gut microbiota and exhibited antibiotic resistance. Further research is needed to determine the effect of gut microbes on M. rotundata health.

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Section: (B) Fungal Community Diversitymentioning

confidence: 95%

Interactions between fungi and bacteria influence microbial community structure in the Megachile rotundata larval gut

2014

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“…A subset of the gut microbiota has been reported to be shared among host individuals within various animal species, including Anopheles mosquitoes, the honey bee Apis mellifera, zebrafish Danio rerio and the laboratory mouse (Mohr and Tebbe, 2006;Martinson et al, 2011;Roeselers et al, 2011;Wang et al, 2011;Pedron et al, 2012;Tang et al, 2012). This subset has been described as the core microbiota (Hamady and Knight, 2009;Shade and Handelsman, 2012).…”

Section: Introductionmentioning

confidence: 99%

The inconstant gut microbiota of Drosophila species revealed by 16S rRNA gene analysis

2013

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The gut microorganisms in some animals are reported to include a core microbiota of consistently associated bacteria that is ecologically distinctive and may have coevolved with the host. The core microbiota is promoted by positive interactions among bacteria, favoring shared persistence; its retention over evolutionary timescales is evident as congruence between host phylogeny and bacterial community composition. This study applied multiple analyses to investigate variation in the composition of gut microbiota in drosophilid flies. First, the prevalence of five previously described gut bacteria (Acetobacter and Lactobacillus species) in individual flies of 21 strains (10 Drosophila species) were determined. Most bacteria were not present in all individuals of most strains, and bacterial species pairs co-occurred in individual flies less frequently than predicted by chance, contrary to expectations of a core microbiota. A complementary pyrosequencing analysis of 16S rRNA gene amplicons from the gut microbiota of 11 Drosophila species identified 209 bacterial operational taxonomic units (OTUs), with near-saturating sampling of sequences, but none of the OTUs was common to all host species. Furthermore, in both of two independent sets of Drosophila species, the gut bacterial community composition was not congruent with host phylogeny. The final analysis identified no common OTUs across three wild and four laboratory samples of D. melanogaster. Our results yielded no consistent evidence for a core microbiota in Drosophila. We conclude that the taxonomic composition of gut microbiota varies widely within and among Drosophila populations and species. This is reminiscent of the patterns of bacterial composition in guts of some other animals, including humans.

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“…Here we applied this new technology to directly survey the genetic composition of density gradient fractions and identify metabolically active bacteria, which gave enhanced sensitivity compared with the gel electrophoresis-based methods, and facilitated subsequent phylogenic classification. By comparing equivalent heavy fractions from both the 13 C-labeled and 12 C-control samples, we find that Pantoea and Enterococcus became labeled upon isotope probing. Once the metabolically active bacteria have been identified, other analysis such as the fluorescence in situ hybridization (FISH) using specific probes and metagenomic analysis of the labeled DNA recovered from active community members can be conducted to provide a comprehensive insight into the true symbionts associated with the host.…”

Section: Discussionmentioning

confidence: 97%

“…Moreover, the results are independent from isolation and cultivation of the gut harbored bacteria as in the past. Almost 99% of bacteria are not cultivable and the simulation of the environmental conditions prevailing in the gut is difficult 12 . By using PCR, 16S rRNA gene fragments (a widely used phylogenetic gene marker among bacteria) could be selectively amplified from a mixed DNA template of gut bacterial communities, sequenced, and cloned.…”

Section: Introductionmentioning

confidence: 99%

Identification of Metabolically Active Bacteria in the Gut of the Generalist <em>Spodoptera littoralis</em> via DNA Stable Isotope Probing Using <sup>13</sup>C-Glucose

Shao

Arias-Cordero

Boland

2013

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Guts of most insects are inhabited by complex communities of symbiotic nonpathogenic bacteria. Within such microbial communities it is possible to identify commensal or mutualistic bacteria species. The latter ones, have been observed to serve multiple functions to the insect, i.e. helping in insect reproduction 1 , boosting the immune response 2 , pheromone production 3 , as well as nutrition, including the synthesis of essential amino acids 4, among others.Due to the importance of these associations, many efforts have been made to characterize the communities down to the individual members. However, most of these efforts were either based on cultivation methods or relied on the generation of 16S rRNA gene fragments which were sequenced for final identification. Unfortunately, these approaches only identified the bacterial species present in the gut and provided no information on the metabolic activity of the microorganisms.To characterize the metabolically active bacterial species in the gut of an insect, we used stable isotope probing (SIP) in vivo employing 13 Cglucose as a universal substrate. This is a promising culture-free technique that allows the linkage of microbial phylogenies to their particular metabolic activity. This is possible by tracking stable, isotope labeled atoms from substrates into microbial biomarkers, such as DNA and RNA 5 . The incorporation of 13 C isotopes into DNA increases the density of the labeled DNA compared to the unlabeled ( 12 C) one. In the end, the 13 Clabeled DNA or RNA is separated by density-gradient ultracentrifugation from the 12 C-unlabeled similar one 6 . Subsequent molecular analysis of the separated nucleic acid isotopomers provides the connection between metabolic activity and identity of the species.Here, we present the protocol used to characterize the metabolically active bacteria in the gut of a generalist insect (our model system), Spodoptera littoralis (Lepidoptera, Noctuidae). The phylogenetic analysis of the DNA was done using pyrosequencing, which allowed high resolution and precision in the identification of insect gut bacterial community. As main substrate, 13 C-labeled glucose was used in the experiments. The substrate was fed to the insects using an artificial diet. Video LinkThe video component of this article can be found at

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Complexity and Variability of Gut Commensal Microbiota in Polyphagous Lepidopteran Larvae

Cited by 172 publications

References 43 publications

Interactions between fungi and bacteria influence microbial community structure in the Megachile rotundata larval gut

Interactions between fungi and bacteria influence microbial community structure in the Megachile rotundata larval gut

The inconstant gut microbiota of Drosophila species revealed by 16S rRNA gene analysis

Identification of Metabolically Active Bacteria in the Gut of the Generalist <em>Spodoptera littoralis</em> via DNA Stable Isotope Probing Using <sup>13</sup>C-Glucose

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