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Variation in honey bee gut microbial diversity affected by ontogenetic stage, age and geographic location.

Hroncova Z, Havlik J, Killer J, Doskocil I, Tyl J, Kamler M, Titera D, Hakl J, Mrazek J, Bunesova V, Rada V - PLoS ONE (2015)

Bottom Line: However, during pupation, microbial counts were significantly reduced but recovered quickly by 6 days post-emergence.The results suggest that 3-day 4th instar larvae contain low microbial counts that increase 2-fold by day 6 and then decrease during pupation.We found that bacterial counts do not show only yearly cycles within a colony, but vary on the individual level.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague, Czech Republic.

ABSTRACT
Social honey bees, Apis mellifera, host a set of distinct microbiota, which is similar across the continents and various honey bee species. Some of these bacteria, such as lactobacilli, have been linked to immunity and defence against pathogens. Pathogen defence is crucial, particularly in larval stages, as many pathogens affect the brood. However, information on larval microbiota is conflicting. Seven developmental stages and drones were sampled from 3 colonies at each of the 4 geographic locations of A. mellifera carnica, and the samples were maintained separately for analysis. We analysed the variation and abundance of important bacterial groups and taxa in the collected bees. Major bacterial groups were evaluated over the entire life of honey bee individuals, where digestive tracts of same aged bees were sampled in the course of time. The results showed that the microbial tract of 6-day-old 5th instar larvae were nearly equally rich in total microbial counts per total digestive tract weight as foraging bees, showing a high percentage of various lactobacilli (Firmicutes) and Gilliamella apicola (Gammaproteobacteria 1). However, during pupation, microbial counts were significantly reduced but recovered quickly by 6 days post-emergence. Between emergence and day 6, imago reached the highest counts of Firmicutes and Gammaproteobacteria, which then gradually declined with bee age. Redundancy analysis conducted using denaturing gradient gel electrophoresis identified bacterial species that were characteristic of each developmental stage. The results suggest that 3-day 4th instar larvae contain low microbial counts that increase 2-fold by day 6 and then decrease during pupation. Microbial succession of the imago begins soon after emergence. We found that bacterial counts do not show only yearly cycles within a colony, but vary on the individual level. Sampling and pooling adult bees or 6th day larvae may lead to high errors and variability, as both of these stages may be undergoing dynamic succession.

No MeSH data available.


Related in: MedlinePlus

Biplot from redundancy analysis (RDA) explaining the distribution of honey bee ontogenetic stages according to major bacterial strain abundance.Test of interactions between factors location and ontogenetic stage: A, crude data considering absolute DGGE band intensities; B, centred and standardized data considering relative band intensities. Abbreviations: Gil, Gilliamella apicola; Sno, Snodgrassella alvi; Lac, Lactobacillus sp.; Rhi, Rhizobiales bacterium; Fri, Frischella perrara, UM, unknown multiple—probable DNA heterodimer. For further strain descriptions, see S5 Fig. Dotted shapes surrounding each ontogenetic stage were created as an aid in visualization. Eighteen bacterial strains occurring as major 16S rDNA DGGE bands were used for statistical analysis, while only selected strains are plotted as arrows. Same descriptions are for bands of the same sequence occurring at multiple locations of the line. Blue arrows show hypothetical developmental timeline. Its dotted part is ambiguous.
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pone.0118707.g002: Biplot from redundancy analysis (RDA) explaining the distribution of honey bee ontogenetic stages according to major bacterial strain abundance.Test of interactions between factors location and ontogenetic stage: A, crude data considering absolute DGGE band intensities; B, centred and standardized data considering relative band intensities. Abbreviations: Gil, Gilliamella apicola; Sno, Snodgrassella alvi; Lac, Lactobacillus sp.; Rhi, Rhizobiales bacterium; Fri, Frischella perrara, UM, unknown multiple—probable DNA heterodimer. For further strain descriptions, see S5 Fig. Dotted shapes surrounding each ontogenetic stage were created as an aid in visualization. Eighteen bacterial strains occurring as major 16S rDNA DGGE bands were used for statistical analysis, while only selected strains are plotted as arrows. Same descriptions are for bands of the same sequence occurring at multiple locations of the line. Blue arrows show hypothetical developmental timeline. Its dotted part is ambiguous.

Mentions: In Fig. 2A, the first canonical axis (horizontal) explained 15.3% (i.e. 35% of total explained) of strain variability and separated stages according to age with L1, L3, PW and PB on the left, L6 in the middle of the plot and adult stages mainly on the right side of the plot. The abundance of nearly all strains clearly increased with age (towards the right side). The highest variability according to locality was observed within drones and young bees, where the total strain abundance of these stages may be more similar to adults (e.g. drones in Dol) or larval stages (e.g. drones in Ustrasice). The data distribution for the second canonical axes (vertical) showed a trend in the differences within adult stages with drones in the lower plot, young bees in the middle and foraging bees in the upper part of the plot. Lactobacillus Lac 3, Lac 4, Snodgrassella alvi (Betaproteobacteria-Beta) and unknown Rhizobiales bacterium (Rhi) (Alphaproteobacteria-Alpha) appeared to be strains that were generally characteristic for foraging bees, while Gilliamella strains were characteristic for the microbiota of young bees, particularly drones. Although the samples were pooled, young bees at some locations appear to be less populated by distinct microbiota and are thus appear on the left side of the first canonical axis with larvae and pupae. This might be to the fact, that microbial successions in young bees may be faster at some sites or colonies compared to others.


Variation in honey bee gut microbial diversity affected by ontogenetic stage, age and geographic location.

Hroncova Z, Havlik J, Killer J, Doskocil I, Tyl J, Kamler M, Titera D, Hakl J, Mrazek J, Bunesova V, Rada V - PLoS ONE (2015)

Biplot from redundancy analysis (RDA) explaining the distribution of honey bee ontogenetic stages according to major bacterial strain abundance.Test of interactions between factors location and ontogenetic stage: A, crude data considering absolute DGGE band intensities; B, centred and standardized data considering relative band intensities. Abbreviations: Gil, Gilliamella apicola; Sno, Snodgrassella alvi; Lac, Lactobacillus sp.; Rhi, Rhizobiales bacterium; Fri, Frischella perrara, UM, unknown multiple—probable DNA heterodimer. For further strain descriptions, see S5 Fig. Dotted shapes surrounding each ontogenetic stage were created as an aid in visualization. Eighteen bacterial strains occurring as major 16S rDNA DGGE bands were used for statistical analysis, while only selected strains are plotted as arrows. Same descriptions are for bands of the same sequence occurring at multiple locations of the line. Blue arrows show hypothetical developmental timeline. Its dotted part is ambiguous.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4358834&req=5

pone.0118707.g002: Biplot from redundancy analysis (RDA) explaining the distribution of honey bee ontogenetic stages according to major bacterial strain abundance.Test of interactions between factors location and ontogenetic stage: A, crude data considering absolute DGGE band intensities; B, centred and standardized data considering relative band intensities. Abbreviations: Gil, Gilliamella apicola; Sno, Snodgrassella alvi; Lac, Lactobacillus sp.; Rhi, Rhizobiales bacterium; Fri, Frischella perrara, UM, unknown multiple—probable DNA heterodimer. For further strain descriptions, see S5 Fig. Dotted shapes surrounding each ontogenetic stage were created as an aid in visualization. Eighteen bacterial strains occurring as major 16S rDNA DGGE bands were used for statistical analysis, while only selected strains are plotted as arrows. Same descriptions are for bands of the same sequence occurring at multiple locations of the line. Blue arrows show hypothetical developmental timeline. Its dotted part is ambiguous.
Mentions: In Fig. 2A, the first canonical axis (horizontal) explained 15.3% (i.e. 35% of total explained) of strain variability and separated stages according to age with L1, L3, PW and PB on the left, L6 in the middle of the plot and adult stages mainly on the right side of the plot. The abundance of nearly all strains clearly increased with age (towards the right side). The highest variability according to locality was observed within drones and young bees, where the total strain abundance of these stages may be more similar to adults (e.g. drones in Dol) or larval stages (e.g. drones in Ustrasice). The data distribution for the second canonical axes (vertical) showed a trend in the differences within adult stages with drones in the lower plot, young bees in the middle and foraging bees in the upper part of the plot. Lactobacillus Lac 3, Lac 4, Snodgrassella alvi (Betaproteobacteria-Beta) and unknown Rhizobiales bacterium (Rhi) (Alphaproteobacteria-Alpha) appeared to be strains that were generally characteristic for foraging bees, while Gilliamella strains were characteristic for the microbiota of young bees, particularly drones. Although the samples were pooled, young bees at some locations appear to be less populated by distinct microbiota and are thus appear on the left side of the first canonical axis with larvae and pupae. This might be to the fact, that microbial successions in young bees may be faster at some sites or colonies compared to others.

Bottom Line: However, during pupation, microbial counts were significantly reduced but recovered quickly by 6 days post-emergence.The results suggest that 3-day 4th instar larvae contain low microbial counts that increase 2-fold by day 6 and then decrease during pupation.We found that bacterial counts do not show only yearly cycles within a colony, but vary on the individual level.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague, Czech Republic.

ABSTRACT
Social honey bees, Apis mellifera, host a set of distinct microbiota, which is similar across the continents and various honey bee species. Some of these bacteria, such as lactobacilli, have been linked to immunity and defence against pathogens. Pathogen defence is crucial, particularly in larval stages, as many pathogens affect the brood. However, information on larval microbiota is conflicting. Seven developmental stages and drones were sampled from 3 colonies at each of the 4 geographic locations of A. mellifera carnica, and the samples were maintained separately for analysis. We analysed the variation and abundance of important bacterial groups and taxa in the collected bees. Major bacterial groups were evaluated over the entire life of honey bee individuals, where digestive tracts of same aged bees were sampled in the course of time. The results showed that the microbial tract of 6-day-old 5th instar larvae were nearly equally rich in total microbial counts per total digestive tract weight as foraging bees, showing a high percentage of various lactobacilli (Firmicutes) and Gilliamella apicola (Gammaproteobacteria 1). However, during pupation, microbial counts were significantly reduced but recovered quickly by 6 days post-emergence. Between emergence and day 6, imago reached the highest counts of Firmicutes and Gammaproteobacteria, which then gradually declined with bee age. Redundancy analysis conducted using denaturing gradient gel electrophoresis identified bacterial species that were characteristic of each developmental stage. The results suggest that 3-day 4th instar larvae contain low microbial counts that increase 2-fold by day 6 and then decrease during pupation. Microbial succession of the imago begins soon after emergence. We found that bacterial counts do not show only yearly cycles within a colony, but vary on the individual level. Sampling and pooling adult bees or 6th day larvae may lead to high errors and variability, as both of these stages may be undergoing dynamic succession.

No MeSH data available.


Related in: MedlinePlus