# Environmental Transmission of Symbionts in the Mangrove Crabs Aratus pisonii and Minuca rapax: Acquisition of the Bacterial Community through Larval Development to Juvenile Stage

## TL;DR

This study shows that two mangrove crabs acquire their gill bacteria from the environment, not from their parents, as no bacteria were found in eggs or larvae.

## Contribution

The study provides evidence for environmental acquisition of bacterial symbionts in crabs during larval development.

## Key findings

- Bacteria were not detected in crab gonads, eggs, or larvae, indicating no vertical transmission.
- Juvenile crabs of both species had gill bacteria similar to those in adults, suggesting environmental acquisition.
- Bacterial communities in juveniles were distinct between species and not shared despite co-occurring habitats.

## Abstract

Aratus pisonii and Minuca rapax are two brachyuran crabs living with bacterial ectosymbionts located on gill lamellae. One previous study has shown that several rod-shaped bacterial morphotypes are present and the community is dominated by Alphaproteobacteria and Bacteroidota. This study aims to identify the mode of transmission of the symbionts to the new host generations and to identify the bacterial community colonizing the gills of juveniles. We tested for the presence of bacteria using PCR with universal primers targeting the 16S rRNA encoding gene from gonads, eggs, and different larval stages either obtained in laboratory conditions or from the field. The presence of bacteria on juvenile gills was also characterized by scanning electron microscopy, and subsequently identified by metabarcoding analysis. Gonads, eggs, and larvae were negative to PCR tests, suggesting that bacteria are not present at these stages in significant densities. On the other hand, juveniles of both species display three rod-shaped bacterial morphotypes on gill lamellae, and sequencing revealed that the community is dominated by Bacteroidota and Alphaproteobacteria on A. pisonii juveniles, and by Alphaprotobacteria, Bacteroidota, and Acidimicrobia on M. rapax juveniles. Despite the fact that juveniles of both species co-occur in the same biotope, no shared bacterial phylotype was identified. However, some of the most abundant bacteria present in adults are also present in juveniles of the same species, suggesting that juvenile-associated communities resemble those of adults. Because some of these bacteria were also found in crab burrow water, we hypothesize that the bacterial community is established gradually during the life of the crab starting from the megalopa stage and involves epibiosis-competent bacteria that occur in the environment.