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Why bilaterally symmetrical organisms express handedness remains an important question in evolutionary biology. In some species, anatomical asymmetries have evolved that accompany behavioral handedness, yet we know remarkably little about causal links between asymmetric morphological traits and behavior. Here, we explore if a dextral or sinistral orientation of the male intromittent organ predicts side preferences in male behaviors. Our study addresses this question in the Costa Rican livebearing fish, Xenophallus umbratilis. This fish has a bilaterally symmetrical body plan, with one exception—the male anal fin (gonopodium), used to inseminate females, terminates with a distinct left- or right-handed corkscrew morphology. We used a detour assay to test males for side biases in approach behavior when exposed to four different stimuli (predator, potential mate, novel object, empty tank control). We found that left morph males preferred using their right eye to view potential mates, predators, and the control, and that right morph males preferred to use their left eye to view potential mates and predators, and their right eye to view the control. Males of both morphs displayed no eye bias when approaching the novel object. Our results suggest that there is a strong link between behavior and gonopodium orientation, with right and left morph males responding with opposite directional behaviors when presented with the same stimuli. This presents the intriguing possibility that mating preferences—in this case constrained by gonopodial morphology—could be driving lateralized decision making in a variety of non-mating behaviors.

Species delimitation among closely related species is challenging because traditional phenotype-based approaches, e.g., morphology, ecological, or chemical characteristics, often produce conflicting results. With the advent of high-throughput sequencing, it has become increasingly cost-effective to acquire genome-scale data which can resolve previously ambiguous species boundaries. As the availability of genome-scale data has increased, numerous species delimitation analyses, such as BPP and SNAPP+Bayes factor delimitation (BFD*), have been developed to delimit species boundaries. However, even empirical molecular species delimitation approaches can be biased by confounding evolutionary factors, e.g., hybridization/introgression and incomplete lineage sorting, and computational limitations. Here we investigate species boundaries and the potential for micro-endemism in a lineage of lichen-forming fungi, Niebla Rundel & Bowler in the family Ramalinaceae. The species delimitation models tend to support more specious groupings, but were unable to infer robust, consistent species delimitations. The results of our study highlight the problem of delimiting species, particularly in groups such as Niebla, with complex, recent phylogeographic histories.

Xenophallus umbratilis is a freshwater livebearing fish that exhibits unique antisymmetry in the male gonopodium, which terminates in either a dextral or sinistral twist. This asymmetry in the gonopodium suggests that males might exhibit side-biased behavior when interacting with females to mate. We conducted two assays to assess the laterality of male and female mating interactions based on gonopodial morphology. We observed lateralized mating behavior in one test where males with sinistral gonopodial morphology interacted with a single female. However, we did not find lateralized mating behavior in males with dextral gonopodial morphology. We also examined male and female positioning in trials that placed a single female with five males, all with the same morphology. These trials also showed no evidence of lateralized body positioning.

Antisymmetry is a striking, yet puzzling form of biological asymmetry. The livebearing fish Xenophallus umbratilis exhibits antisymmetry in the male intromittent organ and provides a system that is well-suited for studying the nature of variation in antisymmetrical traits. Using geometric morphometrics, we test the hypothesis that because the gonopodium is critical to fitness there will not be significant differences in gonopodium shape between the two gonopodial morphs in this species. Our results are consistent with this prediction, though we found that gonopodium shape differed with gonopodium size.