The role of the gut microbiota in the dietary niche expansion of fishing bats.

BACKGROUND: Due to its central role in animal nutrition, the gut microbiota is likely a relevant factor shaping dietary niche shifts. We analysed both the impact and contribution of the gut microbiota to the dietary niche expansion of the only four bat species that have incorporated fish into their primarily arthropodophage diet. RESULTS: We first compared the taxonomic and functional features of the gut microbiota of the four piscivorous bats to that of 11 strictly arthropodophagous species using 16S rRNA targeted amplicon sequencing. Second, we increased the resolution of our analyses for one of the piscivorous bat species, namely Myotis capaccinii, and analysed multiple populations combining targeted approaches with shotgun sequencing. To better understand the origin of gut microorganisms, we also analysed the gut microbiota of their fish prey (Gambusia holbrooki). Our analyses showed that piscivorous bats carry a characteristic gut microbiota that differs from that of their strict arthropodophagous counterparts, in which the most relevant bacteria have been directly acquired from their fish prey. This characteristic microbiota exhibits enrichment of genes involved in vitamin biosynthesis, as well as complex carbohydrate and lipid metabolism, likely providing their hosts with an enhanced capacity to metabolise the glycosphingolipids and long-chain fatty acids that are particularly abundant in fish. CONCLUSIONS: Our results depict the gut microbiota as a relevant element in facilitating the dietary transition from arthropodophagy to piscivory.

Synergetic effects of immune challenge and stress depress cortisol, inflammatory response and antioxidant activity in fish-eating Myotis.

One of the most common tools in conservation physiology is the assessment of environmental stress via glucocorticoid measurement. However, little is known of its relationship with other stress-related biomarkers, and how the incidence of an immune challenge during long-term stress could affect an individual's overall stress response. We investigated here the relationship between basal and post-acute stress fecal cortisol metabolite (FC) with different antioxidant enzymes, oxidative damage and immune parameters in the fish-eating bat, Myotis vivesi We found that in both basal and post-stress conditions, FC was highly related with a number of antioxidant enzymes and immune parameters, but not to oxidative damage. We also assessed changes of FC through the seasons. Basal FC samples and stress reactivity after short-duration stress displayed similar levels during summer, autumn and early winter, but lower concentrations in late winter. Stress reactivity after long-duration stress was greater in summer and early winter. Finally, we tested the effect of a simultaneous exposure to a long, strong stress stimulus with an immune response stimulation by administrating adrenocorticotropic hormone (ACTH) and phytohemagglutinin (PHA) after 42 h. Results showed that when both stimuli were administrated, FC concentrations, inflammation and some antioxidant activity were lowered in comparison with the control and individual administration of the challenges. Our findings support the idea that animals maintain constant basal glucocorticoid levels when living in challenging environments, but response to acute stress differs seasonally and immune defense mechanisms and stress responses might be compromised when confronted with multiple challenges.

Acoustic evaluation of behavioral states predicted from GPS tracking: a case study of a marine fishing bat.

BACKGROUND: Multiple methods have been developed to infer behavioral states from animal movement data, but rarely has their accuracy been assessed from independent evidence, especially for location data sampled with high temporal resolution. Here we evaluate the performance of behavioral segmentation methods using acoustic recordings that monitor prey capture attempts. METHODS: We recorded GPS locations and ultrasonic audio during the foraging trips of 11 Mexican fish-eating bats, Myotis vivesi, using miniature bio-loggers. We then applied five different segmentation algorithms (k-means clustering, expectation-maximization and binary clustering, first-passage time, hidden Markov models, and correlated velocity change point analysis) to infer two behavioral states, foraging and commuting, from the GPS data. To evaluate the inference, we independently identified characteristic patterns of biosonar calls ("feeding buzzes") that occur during foraging in the audio recordings. We then compared segmentation methods on how well they correctly identified the two behaviors and if their estimates of foraging movement parameters matched those for locations with buzzes. RESULTS: While the five methods differed in the median percentage of buzzes occurring during predicted foraging events, or true positive rate (44-75%), a two-state hidden Markov model had the highest median balanced accuracy (67%). Hidden Markov models and first-passage time predicted foraging flight speeds and turn angles similar to those measured at locations with feeding buzzes and did not differ in the number or duration of predicted foraging events. CONCLUSION: The hidden Markov model method performed best at identifying fish-eating bat foraging segments; however, first-passage time was not significantly different and gave similar parameter estimates. This is the first attempt to evaluate segmentation methodologies in echolocating bats and provides an evaluation framework that can be used on other species.

Bite force, cranial morphometrics and size in the fishing bat Myotis vivesi (Chiroptera: Vespertilionidae)

Abstract Fish-eating in bats evolved independently in Myotis vivesi (Vespertillionidae) and Noctilio leporinus (Noctilionidae). We compared cranial morphological characters and bite force between these species to test the existence of evolutionary parallelism in piscivory. We collected cranial distances of M. vivesi, two related insectivorous bats (M. velifer and M. keaysi), two facultatively piscivorous bats (M. daubentonii and M. capaccinii), and N. leporinus. We analyzed morphometric data applying multivariate methods to test for differences among the six species. We also measured bite force in M. vivesi and evaluated if this value was well predicted by its cranial size. Both piscivorous species were morphologically different from the facultatively piscivorous and insectivorous species, and skull size had a significant contribution to this difference. However, we did not find morphological and functional similarities that could be interpreted as parallelisms between M. vivesi and N. leporinus. These two piscivorous species differed significantly in cranial measurements and in bite force. Bite force measured for M. vivesi was well predicted by skull size. Piscivory in M. vivesi might be associated to the existence of a vertically displaced temporal muscle and an increase in gape angle that allows a moderate bite force to process food.(AU)

Resumen La alimentación por peces en murciélagos evolucionó independientemente en Myotis vivesi (Vespertilionidae) y Noctilio leporinus (Phyllostomidae). En este estudio se compararon características craneales morfológicas y fuerza de mordida entre estas especies, para probar la existencia de paralelismo evolucionario en piscivoría. Se recolectaron distancias craneales en M. vivesi, dos parientes insectívoros (M. velifer y M. keaysi), dos murciélagos piscívoros facultativos (M. daubentonii y M. capaccinii), y N. leporinus. Se analizaron datos morfométricos aplicando múltiples métodos para probar las diferencias entre las seis especies. Se midió la fuerza de mordida en M. vivesi y se evalúo si puede ser predicha por el tamaño del cráneo. Las especies piscívoras fueron morfológicamente diferentes de las facultativamente piscívoras y las insectívoras, el tamaño del cráneo tuvo una contribución significativa en esta diferencia. Sin embargo, no encontramos semejanzas morfológicas y funcionales que puedan ser interpretadas como paralelismos entre M. vivesi y N. leporinus. Estas dos especies piscívoras difieren significativamente en medidas craneales y fuerza de mordida. La fuerza de mordida en M. vivesi fue efectivamente predicha por el tamaño de cráneo. La piscivoría en M. vivesi puede estar asociada con la existencia de un músculo temporal verticalmente desplazado y el incremento en el ángulo de apertura mandibular que permite moderar la fuerza de mordida para procesar el alimento.(AU)

Resource Ephemerality Drives Social Foraging in Bats.

Observations of animals feeding in aggregations are often interpreted as events of social foraging, but it can be difficult to determine whether the animals arrived at the foraging sites after collective search [1-4] or whether they found the sites by following a leader [5, 6] or even independently, aggregating as an artifact of food availability [7, 8]. Distinguishing between these explanations is important, because functionally, they might have very different consequences. In the first case, the animals could benefit from the presence of conspecifics, whereas in the second and third, they often suffer from increased competition [3, 9-13]. Using novel miniature sensors, we recorded GPS tracks and audio of five species of bats, monitoring their movement and interactions with conspecifics, which could be inferred from the audio recordings. We examined the hypothesis that food distribution plays a key role in determining social foraging patterns [14-16]. Specifically, this hypothesis predicts that searching for an ephemeral resource (whose distribution in time or space is hard to predict) is more likely to favor social foraging [10, 13-15] than searching for a predictable resource. The movement and social interactions differed between bats foraging on ephemeral versus predictable resources. Ephemeral species changed foraging sites and showed large temporal variation nightly. They aggregated with conspecifics as was supported by playback experiments and computer simulations. In contrast, predictable species were never observed near conspecifics and showed high spatial fidelity to the same foraging sites over multiple nights. Our results suggest that resource (un)predictability influences the costs and benefits of social foraging.

Baseline and post-stress seasonal changes in immunocompetence and redox state maintenance in the fishing bat Myotis vivesi.

Little is known of how the stress response varies when animals confront seasonal life-history processes. Antioxidant defenses and damage caused by oxidative stress and their link with immunocompetence are powerful biomarkers to assess animal´s physiological stress response. The aim of this study was A) to determine redox state and variation in basal (pre-acute stress) immune function during summer, autumn and winter (spring was not assessed due to restrictions in collecting permit) in the fish-eating Myotis (Myotis vivesi; Chiroptera), and B) to determine the effect of acute stress on immunocompetence and redox state during each season. Acute stress was stimulated by restricting animal movement for 6 and 12 h. The magnitude of the cellular immune response was higher during winter whilst that of the humoral response was at its highest during summer. Humoral response increased after 6 h of movement restriction stress and returned to baseline levels after 12 h. Basal redox state was maintained throughout the year, with no significant changes in protein damage, and antioxidant activity was modulated mainly in relation to variation to environment cues, increasing during high temperatures and decreasing during windy nights. Antioxidant activity increased after the 6 h of stressful stimuli especially during summer and autumn, and to a lesser extent in early winter, but redox state did not vary. However, protein damage increased after 12 h of stress during summer. Prolonged stress when the bat is engaged in activities of high energy demand overcame its capacity to maintain homeostasis resulting in oxidative damage.

Metabolic Cost of the Activation of Immune Response in the Fish-Eating Myotis (Myotis vivesi): The Effects of Inflammation and the Acute Phase Response.

Inflammation and activation of the acute phase response (APR) are energetically demanding processes that protect against pathogens. Phytohaemagglutinin (PHA) and lipopolysaccharide (LPS) are antigens commonly used to stimulate inflammation and the APR, respectively. We tested the hypothesis that the APR after an LPS challenge was energetically more costly than the inflammatory response after a PHA challenge in the fish-eating Myotis bat (Myotis vivesi). We measured resting metabolic rate (RMR) after bats were administered PHA and LPS. We also measured skin temperature (Tskin) after the LPS challenge and skin swelling after the PHA challenge. Injection of PHA elicited swelling that lasted for several days but changes in RMR and body mass were not significant. LPS injection produced a significant increase in Tskin and in RMR, and significant body mass loss. RMR after LPS injection increased by 140-185% and the total cost of the response was 6.50 kJ. Inflammation was an energetically low-cost process but the APR entailed a significant energetic investment. Examination of APR in other bats suggests that the way in which bats deal with infections might not be uniform.

The cost of digestion in the fish-eating myotis (Myotis vivesi).

Flying vertebrates, such as bats, face special challenges with regards to the throughput and digestion of food. On the one hand, as potentially energy-limited organisms, bats must ingest and assimilate energy efficiently in order to satisfy high resting and active metabolic demands. On the other hand, the assimilation of nutrients must be accomplished using a digestive tract that is, compared with that of similarly sized non-flying vertebrates, significantly shorter. Despite these competing demands, and the relative breadth of dietary diversity among bats, little work has been done describing the cost of digestion, termed 'specific dynamic action' (SDA). Here, we provide the first systematic assessment of the SDA response in a bat, the fish-eating myotis (Myotis vivesi). Given the shorter digestive tract and the relatively higher resting and active metabolic rates of bats in general, and based on anecdotal published evidence, we hypothesized that the SDA response in fish-eating myotis would be dependent on meal size and both significantly more brief and intense than in small, non-flying mammals. In agreement with our hypothesis, we found that the peak metabolic rate during digestion, relative to rest, was significantly higher in these bats compared with any other mammals or vertebrates, except for some infrequently eating reptiles and amphibians. Additionally, we found that the magnitude and duration of the SDA response were related to meal size. However, we found that the duration of the SDA response, while generally similar to reported gut transit times in other small bats, was not substantially shorter than in similarly sized non-flying mammals.