A Gastrointestinal Metagenomic Study On Mudskippers Supports Their Terrestrial Adaptation

-Purpose: The adaptability of blue-spotted mudskipper (Boleophthalmus Periophthalmodon; BP) and giant-fin mudskipper (Periophthalmus magnuspinnatus; PM), has been previously reported at the genome level to explain their amphibious life. However, the roles of GI microbiota in their adaptation to the terrestrial life are worth exploring. -Methods: In this study, we mainly utilized metagenomic data from these two representative mudskippers and typical aquicolous fish species to obtain microbial composition, diversity, abundance and potential functions of GI microbiota for comparisons between amphibious and aquicolous fishes. Meanwhile, we summarized the GI microbiota results of representative seawater fishes, freshwater fishes, amphibians, and terrestrial animals by literature mining for comparing those of the mudskippers. -Result: Interestingly the content for each dominant phylum was strikingly different among BP, PM and aquicolous fishes. We also observed that the profile of GI microbiota in mudskippers owned the typical bacterial families for the terrestrial animals, (freshwater and seawater) fishes, and amphibians at the same time, which is consistent with their life style of water-to-land and freshwater to seawater transition. More interestingly, certain bacteria strains like S24-7, previously thought to be specific in terrestrial animals, were also identified in both BP and PM. -Conclusion: The various composite and diversity of mudskipper GI microflora are therefore considered to conduce to their terrestrial adaptation in these amphibious fishes.

More than Breathing Air: Evolutionary Drivers and Physiological Implications of an Amphibious Lifestyle in Fishes

Amphibious and aquatic air-breathing fishes both exchange respiratory gasses with the atmosphere, but these fishes differ in physiology, ecology, and possibly evolutionary origins. We introduce a scoring system to characterize interspecific variation in amphibiousness and use this system to highlight important unanswered questions about the evolutionary physiology of amphibious fishes.

Does leaving water make fish smarter? Terrestrial exposure and exercise improve spatial learning in an amphibious fish

Amphibious fishes transition between aquatic and terrestrial habitats, and must therefore learn to navigate two dramatically different environments. We used the amphibious killifish Kryptolebias marmoratus to test the hypothesis that the spatial learning ability of amphibious fishes would be altered by exposure to terrestrial environments because of neural plasticity in the brain region linked to spatial cognition (dorsolateral pallium). We subjected fish to eight weeks of fluctuating air–water conditions or terrestrial exercise before assessing spatial learning using a bifurcating T-maze, and neurogenesis in the dorsolateral pallium by immunostaining for proliferating cell nuclear antigen. In support of our hypothesis, we found that air–water fluctuations and terrestrial exercise improved some markers of spatial learning. Moreover, air–water and exercised fish had 39% and 46% more proliferating cells in their dorsolateral pallium relative to control fish, respectively. Overall, our findings suggest that fish with more terrestrial tendencies may have a cognitive advantage over those that remain in water, which ultimately may influence their fitness in both aquatic and terrestrial settings. More broadly, understanding the factors that promote neural and behavioural plasticity in extant amphibious fishes may provide insights into how ancestral fishes successfully colonized novel terrestrial environments before giving rise to land-dwelling tetrapods.

Widespread use of emersion and cutaneous ammonia excretion in Aplocheiloid killifishes

The invasion of land required amphibious fishes to evolve new strategies to avoid toxic ammonia accumulation in the absence of water flow over the gills. We investigated amphibious behaviour and nitrogen excretion strategies in six phylogenetically diverse Aplocheiloid killifishes ( Anablepsoides hartii, Cynodonichthys hildebrandi, Rivulus cylindraceus , Kryptolebias marmoratus, Fundulopanchax gardneri , and Aplocheilus lineatus ) in order to determine if a common strategy evolved . All species voluntarily emersed (left water) over several days, and also in response to environmental stressors (low O 2 , high temperature). All species were ammoniotelic in water and released gaseous ammonia (NH 3 volatilization) during air exposure as the primary route for nitrogen excretion. Metabolic depression, urea synthesis, and/or ammonia accumulation during air exposure were not common strategies used by these species. Immunostaining revealed the presence of ammonia-transporting Rhesus proteins (Rhcg1 and Rhcg2) in the skin of all six species, indicating a shared mechanism for ammonia volatilization. We also found Rhcg in the skin of several other fully aquatic fishes, implying that cutaneous ammonia excretion is not exclusive to amphibious fishes. Overall, our results demonstrate that similar nitrogen excretion strategies while out of water were used by all killifish species tested; possibly the result of shared ancestral amphibious traits, phenotypic convergence, or a combination of both.

Out of the frying pan into the air—emersion behaviour and evaporative heat loss in an amphibious mangrove fish ( Kryptolebias marmoratus )

Amphibious fishes often emerse (leave water) when faced with unfavourable water conditions. How amphibious fishes cope with the risks of rising water temperatures may depend, in part, on the plasticity of behavioural mechanisms such as emersion thresholds. We hypothesized that the emersion threshold is reversibly plastic and thus dependent on recent acclimation history rather than on conditions during early development. Kryptolebias marmoratus were reared for 1 year at 25 or 30°C and acclimated as adults (one week) to either 25 or 30°C before exposure to an acute increase in water temperature. The emersion threshold temperature and acute thermal tolerance were significantly increased in adult fish acclimated to 30°C, but rearing temperature had no significant effect. Using a thermal imaging camera, we also showed that emersed fish in a low humidity aerial environment (30°C) lost significantly more heat (3.3°C min −1 ) than those in a high humidity environment (1.6°C min −1 ). In the field, mean relative humidity was 84%. These results provide evidence of behavioural avoidance of high temperatures and the first quantification of evaporative cooling in an amphibious fish. Furthermore, the avoidance response was reversibly plastic, flexibility that may be important for tropical amphibious fishes under increasing pressures from climatic change.