Xenobiotic estradiol-17ß alters gut microbiota of hatchling American alligators (Alligator mississippiensis).

Environmental oestrogens pose serious concerns for ecosystems through their effects on organismal survival and physiology. The gut microbiome is highly vulnerable to environmental influence, yet the effects of oestrogens on gut homeostasis are unknown because they are poorly studied in wildlife populations. To determine the influence of environmental oestrogens (i.e., xenoestrogens) on the diversity and abundance of gut microbiota, we randomly assigned 23 hatchling American alligators (Alligator mississippiensis) to three ecologically relevant treatments (control, low, and high oestrogen concentrations) for 10 weeks. We predicted that xenoestrogen exposure would decrease microbial diversity and abundance within the digestive tract and that this effect would be dose-dependent. Microbial samples were collected following diet treatments and microbial diversity was determined using 16S rRNA gene-sequencing. Individuals in oestrogen-treatment groups had decreased microbial diversity, but a greater relative abundance of operational taxonomic units than those in the control group. In addition, this effect was dose-dependent; as individuals were exposed to more oestrogen, their microbiome became less diverse, less rich and less even. Findings from this study suggest that oestrogen contamination can influence wildlife populations at the internal microbial-level, which may lead to future deleterious health effects.

Male gopher tortoise (Gopherus polyphemus) concentration-dependent social responses to diluted mental gland pheromones.

In complex terrestrial environments, chemical signals can be the most important sensory modality for locating conspecifics for potential mating opportunities, especially in spatially segregated populations or habitats. Organisms must evolve chemical signals to maximize the efficacy of conveying information, particularly in creating trails or mate-choice cues. Long-distance transmission of chemical signals may be an increasingly important management concern for small and fractured populations or potentially threatened species, such as gopher tortoises in the southeastern U.S. Mental gland secretions have been shown to have pheromonal function in gopher tortoises, suggesting a potential role as trail or marking pheromones, allowing males to track females or other males to find females. In this study, male gopher tortoises were given paired presentations of a negative control (distilled water) with serial dilutions (1:4, 1:20, 1:100, and 1:500) of male mental gland secretions. Male tortoises were able to discern treatment differences up to 1:20 diluted secretions, responding with an array of social behaviors (e.g. for the 1:20 dilution trial, carapace alignment and head bobbing occurred more frequently for the mental gland secretion relative to the control; p < 0.01). Multivariate principal components analysis yielded PC1 (including, approach, carapace alignment, head bobbing, tasting air, sniffing, and doubleback) that differed by treatment (p = 0.0007) and also was higher for the 1:20 diluted presentation relative to the 1:500 diluted presentation (p = 0.04). This study provides insight into gopher tortoise ecology, mate-choice, and the utility of environmentally diluted mental gland secretions in the external environment when seeking mating opportunities.

Behavioural discrimination of male mental gland secretions of the gopher tortoise (Gopherus polyphemus) by both sexes.

Chemical communication is important for mate choice, especially at long distances in fragmented populations. The gopher tortoise is a social species that is threatened in the southeast U.S. due to habitat fragmentation and decline. One consequence of habitat loss is reduced mating opportunities, yet chemical signalling in gopher tortoises is relatively under-studied. Here, we investigated chemoreception of tortoise discrimination of chin secretions, or mental gland (MG) secretions. To assess conspecific recognition of male MG secretions, we conducted two paired-choice experiments: one with a neutral odorant control (NC; distilled water) and one with a pungent odorant control (PC; acetone) vs. male MG secretions. Behaviours were defined a priori, and their durations were quantified relative to treatments. Each sex spent significantly more time with MG secretions vs. acetone control during the PC study (p= 0.001). Each sex also sniffed MG swabs more frequently in both studies (PC study: p=0.0003; NC study: p=0.001). A principal components analysis of behavioural durations from the PC study identified one component with a significant treatment effect performed to MG secretions (p=0.0003), including the behaviours sniffing, head bobbing, biting, and eating near a swab. Our study provides the first chemical-behavioural bioassay of MG secretions from male gopher tortoises, suggesting MG secretions may be a source of pheromones.