The microbiome at the interface between environmental stress and animal health: an example from the most threatened vertebrate group.

Nitrate pollution and global warming are ubiquitous stressors likely to interact and affect the health and survival of wildlife, particularly aquatic ectotherms. Animal health is largely influenced by its microbiome (commensal/symbiotic microorganisms), which responds to such stressors. We used a crossed experimental design including three nitrate levels and five temperature regimes to investigate their interactive and individual effects on an aquatic ectotherm, the European common frog. We associated health biomarkers in larvae with changes in gut bacteria diversity and composition. Larvae experienced higher stress levels and lower body condition under high temperatures and nitrate exposure. Developmental rate increased with temperature but decreased with nitrate pollution. Alterations in bacteria composition but not diversity are likely to correlate with the observed outcomes in larvae health. Leucine degradation decreased at higher temperatures corroborating accelerated development, nitrate degradation increased with nitrate level corroborating reduced body condition and an increase in lysine biosynthesis may have helped larvae deal with the combined effects of both stressors. These results reinforce the importance of associating traditional health biomarkers with underlying microbiome changes. Therefore, we urge studies to investigate the effects of environmental stressors on microbiome composition and consequences for host health in a world threatened by biodiversity loss.

Colonization of frog Rana temporaria L. urinary bladder by Gram-negative bacteria leads to decreased effect of arginine-vasotocin on water reabsorption from the urinary bladder.

In frogs and toads the urinary bladder is very important for the maintenance of water balance due to its ability to store water which can be reabsorbed under the action of arginine-vasotocin (AVT). The usage of isolated bladders as a model for studying the osmotic water permeability (OWP) regulation has a disadvantage which relates to high variability of AVT effect among individuals, some showing insensitivity to the hormone. We hypothesized that the response of the bladder to AVT could depend on the colonization of the mucosal epithelium by Gram-negative bacteria. To test this, paired hemibladders of the frog Rana temporaria were used for measurement of OWP and for analysis of Gram-negative bacteria in the bladder tissue or isolated epithelial cells. Among the 206 frogs studied, 41% were infected by different Enterobacteriaceae, with prevalence of Hafnia alvei and Escherichia coli. In infected bladders the basal level of OWP was unchanged, whereas OWP stimulated by AVT was reduced (non-infected: 2.53 ± 0.13, n = 59, infected: 1.21 ± 0.17 µL min(-1) cm(-2), n = 38, for the 15 min of AVT action, P < 0.001). In the sample, 100% of hemibladders that responded to AVT very weakly (OWP <0.5 µL min(-1) cm(-2)) had a bacterial infection. Overnight treatment of hemibladders with mucosal lipopolysaccharide E. coli decreased OWP induced by AVT, forskolin, or IBMX lowering basal and stimulated level of cAMP. The data obtained indicate that the frog bladder epithelium could be colonized by Gram-negative bacteria, probably of cloacal origin, leading to reduction of sensitivity to AVT and to impairment of the urinary bladder to provide osmoregulation.

External application of antibiotic to improve survival of adult laboratory frogs (Rana pipiens).

Survival data for over 500 adult frogs (393 Rana pipiens, 35 Rana clamitans , and 97 Rana temporaria) obtained from commercial suppliers were accumulated. Frogs which developed signs of red-leg disease while being kept in 0.15% saline were cured by addition of appropriate antibiotics to the saline. The appropriate antibiotic was determined by sensitivity testing of bacteria isolated from the sick frogs. When many frogs in a shipment were sick, prophylactic treatment of the entire shipment improved the overall survival in the shipment.