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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

Boxplot of quantitative real-time PCR (qRT-PCR) data of the abundance of selected bacterial groups in pooled samples of total gastrointestinal tract of each honey bee ontogenetic stage.The Y axis shows log-transformed copies of the 16S rRNA gene per gram of honey bee gastrointestinal tract. Boxes show pooled data from 4 locations and 3 hives at each location. 1-, 3- and 6-day old larvae (L1, L3 and L6, respectively), white and black pupae (PW, PB), young bees, drones and flying bees (BY, DR and BF, respectively). The codes of the outliers refer to the location (Pos: Postrizin, Hos: Hostice, Ust: Ustrasice) and colony number (1, 2, 3).
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pone.0118707.g004: Boxplot of quantitative real-time PCR (qRT-PCR) data of the abundance of selected bacterial groups in pooled samples of total gastrointestinal tract of each honey bee ontogenetic stage.The Y axis shows log-transformed copies of the 16S rRNA gene per gram of honey bee gastrointestinal tract. Boxes show pooled data from 4 locations and 3 hives at each location. 1-, 3- and 6-day old larvae (L1, L3 and L6, respectively), white and black pupae (PW, PB), young bees, drones and flying bees (BY, DR and BF, respectively). The codes of the outliers refer to the location (Pos: Postrizin, Hos: Hostice, Ust: Ustrasice) and colony number (1, 2, 3).

Mentions: After defecation and capping, their counts decreased to (3 × 102). During pupation, an interesting rise in Gamma abundance was observed in white pupae samples (8.3 × 107), which was in this case an order of magnitude higher than the mean of the counts observed in EXP1 and rather similar to its maximum value. In black pupae, these bacteria were suppressed and newly emerged bees continued to show very low bacterial counts. The newly emerging bees appeared to be inoculated and bacterial counts of both Gamma and Firm groups within 6 days post-emergence quickly increased to the highest bacterial counts (G:7.9 × 107 vs. F: 1.8 × 109). Shortly after this rapid inoculation, both bacterial groups began decreasing in abundance, primarily in a linear manner as the honey bee aged (Fig. 4). Between 17–24 days after emergence, honey bees become foragers under normal circumstances [38]; both counts of Firm and Gamma were reduced by half compared to young bees at 6 days postemergence. At day 40, counts of Firm were as low as 4 × 108. It is clear that the sampling of young bees of unspecified age for microbiological analyses is linked with large variation error as the bees were in the process of inoculation and rapid succession.


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)

Boxplot of quantitative real-time PCR (qRT-PCR) data of the abundance of selected bacterial groups in pooled samples of total gastrointestinal tract of each honey bee ontogenetic stage.The Y axis shows log-transformed copies of the 16S rRNA gene per gram of honey bee gastrointestinal tract. Boxes show pooled data from 4 locations and 3 hives at each location. 1-, 3- and 6-day old larvae (L1, L3 and L6, respectively), white and black pupae (PW, PB), young bees, drones and flying bees (BY, DR and BF, respectively). The codes of the outliers refer to the location (Pos: Postrizin, Hos: Hostice, Ust: Ustrasice) and colony number (1, 2, 3).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0118707.g004: Boxplot of quantitative real-time PCR (qRT-PCR) data of the abundance of selected bacterial groups in pooled samples of total gastrointestinal tract of each honey bee ontogenetic stage.The Y axis shows log-transformed copies of the 16S rRNA gene per gram of honey bee gastrointestinal tract. Boxes show pooled data from 4 locations and 3 hives at each location. 1-, 3- and 6-day old larvae (L1, L3 and L6, respectively), white and black pupae (PW, PB), young bees, drones and flying bees (BY, DR and BF, respectively). The codes of the outliers refer to the location (Pos: Postrizin, Hos: Hostice, Ust: Ustrasice) and colony number (1, 2, 3).
Mentions: After defecation and capping, their counts decreased to (3 × 102). During pupation, an interesting rise in Gamma abundance was observed in white pupae samples (8.3 × 107), which was in this case an order of magnitude higher than the mean of the counts observed in EXP1 and rather similar to its maximum value. In black pupae, these bacteria were suppressed and newly emerged bees continued to show very low bacterial counts. The newly emerging bees appeared to be inoculated and bacterial counts of both Gamma and Firm groups within 6 days post-emergence quickly increased to the highest bacterial counts (G:7.9 × 107 vs. F: 1.8 × 109). Shortly after this rapid inoculation, both bacterial groups began decreasing in abundance, primarily in a linear manner as the honey bee aged (Fig. 4). Between 17–24 days after emergence, honey bees become foragers under normal circumstances [38]; both counts of Firm and Gamma were reduced by half compared to young bees at 6 days postemergence. At day 40, counts of Firm were as low as 4 × 108. It is clear that the sampling of young bees of unspecified age for microbiological analyses is linked with large variation error as the bees were in the process of inoculation and rapid succession.

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