Standard methods for research on<i>Apis mellifera</i>gut symbionts

Engel, Philipp; James, Rosalind R.; Koga, Ryuichi; Kwong, Waldan K.; McFrederick, Quinn S.; Moran, Nancy A.

doi:10.3896/ibra.1.52.4.07

Cited by 109 publications

(87 citation statements)

References 99 publications

(5 reference statements)

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“…For example, Fructobacillus, a member of the Leuconostocaceae, may produce fermentative products, such as lactic acid, CO 2 , ethanol, or acetate (39); these products may selectively promote the growth of the honey beespecific microbial community. Indeed, selective culture of many of the "core" microbiome members requires elevated atmospheric CO 2 and acidic media such as MRS (40). Importantly, these effects do not require the presence of Fructobacillus in the bee gut but may be mediated by metabolic products ingested by the bee.…”

Section: Figmentioning

confidence: 99%

Interactions between Cooccurring Lactic Acid Bacteria in Honey Bee Hives

Rokop

Horton

Newton

2015

Appl Environ Microbiol

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In contrast to the honey bee gut, which is colonized by a few characteristic bacterial clades, the hive of the honey bee is home to a diverse array of microbes, including many lactic acid bacteria (LAB). In this study, we used culture, combined with sequencing, to sample the LAB communities found across hive environments. Specifically, we sought to use network analysis to identify microbial hubs sharing nearly identical operational taxonomic units, evidence which may indicate cooccurrence of bacteria between environments. In the process, we identified interactions between noncore bacterial members (Fructobacillus and Lactobacillaceae) and honey bee-specific "core" members. Both Fructobacillus and Lactobacillaceae colonize brood cells, bee bread, and nectar and may serve the role of pioneering species, establishing an environment conducive to the inoculation by honey bee core bacteria. Coculture assays showed that these noncore bacterial members promote the growth of honey bee-specific bacterial species. Specifically, Fructobacillus by-products in spent medium supported the growth of the Firm-5 honey bee-specific clade in vitro. Metabolic characterization of Fructobacillus using carbohydrate utilization assays revealed that this strain is capable of utilizing the simple sugars fructose and glucose, as well as the complex plant carbohydrate lignin. We tested Fructobacillus for antibiotic sensitivity and found that this bacterium, which may be important for establishment of the microbiome, is sensitive to the commonly used antibiotic tetracycline. Our results point to the possible significance of "noncore" and environmental microbial community members in the modulation of honey bee microbiome dynamics and suggest that tetracycline use by beekeepers should be limited.

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Section: Figmentioning

confidence: 99%

Interactions between Cooccurring Lactic Acid Bacteria in Honey Bee Hives

Rokop

Horton

Newton

2015

Appl Environ Microbiol

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“…S. alvi grows in a distinct layer adjacent to the gut epithelium, suggesting adherence (12,13) (Fig. 1).…”

Section: Significancementioning

confidence: 99%

“…An abundant constituent of the honey bee and bumble bee gut microbiota is Snodgrassella alvi (10), a member of Betaproteobacteria that is related to human commensals in the genus Neisseria (11). S. alvi colonizes the hindgut ileum, a region analogous to the proximal large intestine in humans and is normally found in contact with the intima lining, of the ileum (12,13).…”

mentioning

confidence: 99%

Genome-wide screen identifies host colonization determinants in a bacterial gut symbiont

Powell

Leonard

Kwong

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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125

152

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Animal guts are often colonized by host-specialized bacterial species to the exclusion of other transient microorganisms, but the genetic basis of colonization ability is largely unknown. The bacterium Snodgrassella alvi is a dominant gut symbiont in honey bees, specialized in colonizing the hindgut epithelium. We developed methods for transposon-based mutagenesis in S. alvi and, using high-throughput DNA sequencing, screened genome-wide transposon insertion (Tnseq) and transcriptome (RNA-seq) libraries to characterize both the essential genome and the genes facilitating host colonization. Comparison of Tn-seq results from laboratory cultures and from monoinoculated worker bees reveal that 519 of 2,226 protein-coding genes in S. alvi are essential in culture, whereas 399 are not essential but are beneficial for gut colonization. Genes facilitating colonization fall into three broad functional categories: extracellular interactions, metabolism, and stress responses. Extracellular components with strong fitness benefits in vivo include trimeric autotransporter adhesins, O antigens, and type IV pili (T4P). Experiments with T4P mutants establish that T4P in S. alvi likely function in attachment and biofilm formation, with knockouts experiencing a competitive disadvantage in vivo. Metabolic processes promoting colonization include essential amino acid biosynthesis and iron acquisition pathways, implying nutrient scarcity within the hindgut environment. Mechanisms to deal with various stressors, such as for the repair of double-stranded DNA breaks and protein quality control, are also critical in vivo. This genome-wide study identifies numerous genetic networks underlying colonization by a gut commensal in its native host environment, including some known from more targeted studies in other hostmicrobe symbioses.type IV pilus | transposon mutagenesis | microbiota | symbiosis | Neisseriaceae

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“…rarefaction curves, the probability of an interspecific encounter (PIE, a diversity index) and Chao1-estimated OTU richness) and beta diversity (microbial diversity across individual guts, i.e. non-metric multi-dimensional scaling (NMDS) of Bray-Curtis dissimilarity matrices), again using the standard methods for honey bee gut symbionts [31] (see online electronic supplementary material for a detailed description). We additionally compared the effects of treatments on bacterial and fungal OTU richness, diversity (PIE), and Chao1-estimated richness (separately) using analysis of variance (ANOVA).…”

Section: (D) Microbial Community Data Analysismentioning

confidence: 99%

“…To utilize the UNITE ITS database [29] and to verify our 16S rDNA analyses, we conducted parallel analyses in the program QIIME [30]. For all analyses we followed the standard methods recommended for honey bee gut symbionts [31]. We analysed the 16S rDNA and ITS data separately, but used similar methods (see the electronic supplementary material for detailed descriptions of our analyses).…”

mentioning

confidence: 99%

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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Standard methods for research onApis melliferagut symbionts

Cited by 109 publications

References 99 publications

Interactions between Cooccurring Lactic Acid Bacteria in Honey Bee Hives

Interactions between Cooccurring Lactic Acid Bacteria in Honey Bee Hives

Genome-wide screen identifies host colonization determinants in a bacterial gut symbiont

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

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