In this study, we characterize the metabolic capabilities of a honey bee larvae-associated bacterium, Bombella apis (formerly Parasaccharibacter apium), and its effects on the nutritional resilience of larvae. We found that B. apis is the only bacterium associated with larvae that can withstand the antimicrobial larval diet. Further, we found that B. apis can synthesize all essential amino acids and significantly alters the amino acid content of synthetic larval diet, largely by increasing the essential amino acid lysine. 

A honey bee symbiont buffers larvae against nutritional stress through lysine supplementation
January 2022

DOI:10.1101/2022.01.11.475899

Methods 
278
 
Culturing bacteria 
Bombella apis A29 was grown at 34°C, ambient oxygen, shaking at 250 rpm, in Bacto-Schmehl 
(BS) liquid media. BS growth media is derived from the larval diet outlined in Schmehl et al., 
2011 conceived for growth of the honey bee larvae and is described in the following section. 
The designation BactoSchmehl is used with the written permission of Dr. Daniel Schmehl.  
Bacto-Schmehl (BS) Media 
- 5% w/v D-Glucose 
- 5% w/v D-Fructose 
- 1% w/v Yeast Extract 
.CC-BY-NC-ND 4.0 International licenseavailable under a
(which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made 
The copyright holder for this preprintthis version posted January 11, 2022. ; https://doi.org/10.1101/2022.01.11.475899doi: bioRxiv preprint 
- 4% v/v 5X Sigma M9 Salt Solution (catalog #M9956) 
- 0.2% v/v Cation solution (1% [v/v]; 100 mM MgSO4 and 10 mM CaCl2 in diH2O) 
- 84.8% Milli-Q H2O 
Cation solution must be autoclaved separate from the M9 salt solution. BS media final pH 6.5.  
Minimal Bacto-Schmehl Media (mBS)  
- D-Glucose 
- D-Fructose 
- 5X Sigma M9 Salt Solution (catalog #M9956) 
- Cation solution (1% [v/v]; 100 mM MgSO4 and 10 mM CaCl2 in diH2O) 
- 1X Sigma MEM Vitamin Solution (catalog #M6895) 
- 1X Sigma MEM Amino Acids solution (catalog #M5550) or 1X Sigma Non-essential Amino Acid Solution (catalog #M7145) 
- Milli-Q H2O 

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