Kristyn E. Sylvia

Sex-specific modulation of the gut microbiome and behavior in Siberian hamsters

The gut microbiome is a diverse, host-specific, and symbiotic bacterial environment that is critical for mammalian survival and exerts a surprising yet powerful influence on brain and behavior. Gut dysbiosis has been linked to a wide range of physical and psychological disorders, including autism spectrum disorders and anxiety, as well as autoimmune and inflammatory disorders. A wealth of information on the effects of dysbiosis on anxiety and depression has been reported in laboratory model systems (e.g., germ-free mice); however, the effects of microbiome disruption on social behaviors (e.g., aggression) of non-model species that may be particularly important in understanding many aspects of physiology and behavior have yet to be fully explored. Here we assessed the sex-specific effects of a broad-spectrum antibiotic on the gut microbiome and its effects on social behaviors in male and female Siberian hamsters (Phodopus sungorus). In Experiment 1, we administered a broad-spectrum antibiotic on a short-term basis and found that antibiotic treatment altered the microbial communities in the gut in male and female hamsters. In Experiment 2, we tested the effects of single versus repeated antibiotic treatment (including a recovery phase) on behavior, and found that two, but not one, treatments caused marked decreases in aggressive behavior, but not other social behaviors, in males; aggression returned to normal levels following recovery. Antibiotic-treated females, in contrast, showed decreased aggression after a single treatment, with all other social behaviors unaffected. Unlike males, female aggression did not return to normal during either recovery period. The present findings demonstrate that modest antibiotic treatment results in marked disruption of the gut microbiome in hamsters, akin to research done in other rodent species and humans. Further, we show that treatment with a broad-spectrum antibiotic, which has dysbiotic effects, also has robust, sex-specific effects on aggression, a critical behavior in the survival and reproductive success of many rodent species.

A gut feeling: Microbiome-brain-immune interactions modulate social and affective behaviors

ABSTRACT The expression of a wide range of social and affective behaviors, including aggression and investigation, as well as anxiety‐ and depressive‐like behaviors, involves interactions among many different physiological systems, including the neuroendocrine and immune systems. Recent work suggests that the gut microbiome may also play a critical role in modulating behavior and likely functions as an important integrator across physiological systems. Microbes within the gut may communicate with the brain via both neural and humoral pathways, providing numerous avenues of research in the area of the gut‐brain axis. We are now just beginning to understand the intricate relationships among the brain, microbiome, and immune system and how they work in concert to influence behavior. The effects of different forms of experience (e.g., changes in diet, immune challenge, and psychological stress) on the brain, gut microbiome, and the immune system have often been studied independently. Though because these systems do not work in isolation, it is essential to shift our focus to the connections among them as we move forward in our investigations of the gut‐brain axis, the shaping of behavioral phenotypes, and the possible clinical implications of these interactions. This review summarizes the recent progress the field has made in understanding the important role the gut microbiome plays in the modulation of social and affective behaviors, as well as some of the intricate mechanisms by which the microbiome may be communicating with the brain and immune system. HighlightsThe gut microbiome may play a critical role in modulating behavioral responses.Gut microbes may communicate with the brain via both neural and humoral pathways.The neuroendocrine system and microbiome interact to influence social behaviors.The gut‐brain‐behavior axis can have important clinical implications.

Overcoming neonatal sickness: Sex-specific effects of sickness on physiology and social behavior

Early-life environmental stressors, including sickness, have the potential to disrupt development in ways that could severely impact fitness. Despite what is known about the effects of sickness on reproduction, the precise physiological mechanisms have not yet been determined. The goal of this study was to investigate the effects of a neonatal immune challenge on adult reproductive physiology and opposite-sex social behavior. Male and female Siberian hamster (Phodopus sungorus) pups were administered lipopolysaccharide ([LPS]; a cell wall component of gram-negative bacteria) or saline injections on postnatal days 3 and 5 and body mass, food intake, and measures of reproductive maturity were taken throughout development. In adulthood, hamsters were placed in staged mating pairs with reproductively mature individuals of the opposite sex, during which a series of behaviors were scored. We found that although males and females showed no change in food intake, body mass, or reproductive behaviors, LPS-treated females had abnormal estrous cycles and smaller ovaries. Females also showed increased investigation of and increased aggression towards males in a reproductive context. In contrast, LPS-treated males showed no change in any physiological measures, nor did they show any changes in behavior. The present findings demonstrate that females may be more robustly affected by neonatal sickness than males and that these effects could have potential impacts on reproductive success. Collectively, the results of this study can be used to expand upon what is already known about sickness and reproduction, specifically the importance of social behaviors involved in pre-copulation and information necessary to choose the appropriate mate.

Sickness-induced changes in physiology do not affect fecundity or same-sex behavior

Previous work in our lab has shown that early-life infection affects female reproductive physiology and function (i.e., smaller ovaries, abnormal estrous cycles) and alters investigation and aggression towards male conspecifics in a reproductive context. Although many studies have investigated the effects of postnatal immune challenge on physiological and behavioral development, fewer studies have examined whether these changes have ultimate effects on reproduction. In the current study, we paired Siberian hamsters (Phodopus sungorus) and simulated a bacterial infection in early life by administering lipopolysaccharide (LPS) to male and female pups on pnd3 and pnd5. In adulthood, hamsters were paired with novel individuals of the same sex, and we scored an array of social behaviors (e.g., investigation, aggression). We then paired animals with individuals of the opposite sex for 5 consecutive nights, providing them with the opportunity to mate. We found that females exhibited impaired reproductive physiology and function in adulthood (i.e., smaller ovaries and abnormal estrous cycles), similar to our previous work. However, both LPS-treated males and females exhibited similar same-sex social behavior when compared with saline-treated controls, they successfully mated, and there were no significant changes in fecundity. These data suggest that the physiological changes in response to neonatal immune challenge may not have long-term effects on reproductive success in a controlled environment. Collectively, the results of this study are particularly important when investigating the relationships between physiology and behavior within an ultimate context. Animals exposed to early-life stress may in fact be capable of compensating for changes in physiology in order to survive and reproduce in some contexts.

A Return to Wisdom: Using Sickness Behaviors to Integrate Ecological and Translational Research

Sickness is typically characterized by fever, anorexia, cachexia, and reductions in social, pleasurable, and sexual behaviors. These responses can be displayed at varying intensities both within and among individuals, and the adaptive nature of sickness responses can be demonstrated by the context-dependent nature of their expression. The study of sickness has become an important area of investigation for researchers in a wide range of areas, including psychoneuroimmunology (PNI) and ecoimmunology (EI). The general goal of PNI is to identify key interactions among the nervous, endocrine and immune systems and behavior, and how disruptions in these processes might contribute to disease states. EI, in turn, has been established more recently within the perspectives of ecology and evolutionary biology, and is aimed more at understanding natural variation in immune function and sickness responses within a broadly integrative, organismal, and evolutionary context. The goal of this review is to examine the literature on sickness from both basic and biomedical perspectives within PNI and EI and to demonstrate how the integrative study of sickness behavior can serve as an integrating agent to connect ecological and translational approaches to the study of disease. By focusing on a set of specific exemplars, including the energetics of sickness, social context, and environmental influences on sickness, we hope to accomplish the larger goal of developing a common synthetic framework to understand sickness from multiple levels of analysis and varying perspectives across the fields of PNI and EI. By applying this integrative approach to sickness, we will be able to develop a more comprehensive view of sickness as a suite of adaptive responses rather than the simply deleterious consequences of illness.

Physiological predictors of leptin vary during menses and ovulation in healthy women

Although research has shown interactions between the reproductive system and energy homeostasis, it is not clear how environmental or behavioral factors may factor into these associations. Here we aimed to determine how changes in reproductive state (i.e., phase of the menstrual cycle) and other behavioral and physiological factors may influence leptin levels in healthy women, as well as how sexual activity may play a role in leptin modulation. We collected serum and saliva from 32 healthy women and measured leptin, estradiol, and progesterone. Participants also completed surveys of demographics, health and sexual behaviors, and physical activity. Leptin was predicted by meals per day and missed meals at both menses and ovulation. However, estradiol and physical activity were stronger predictors of leptin at menses, while sexual activity was a stronger predictor of leptin at ovulation. These findings suggest that predictors of serum leptin, and possibly energy storage and expenditure, vary across the menstrual cycle.

Acute intraperitoneal lipopolysaccharide influences the immune system in the absence of gut dysbiosis

There is bidirectional communication between the immune system and the gut microbiome, however the precise mechanisms regulating this crosstalk are not well understood. Microbial‐associated molecular patterns (MAMPs) within the gut, including lipopolysaccharide (LPS) that produces a quick and robust activation of the immune system, may be one way by which these interactions occur. Endogenous levels of LPS in the gut are low enough that they do not usually cause disease, although, in times of increased LPS loads, they may be capable of increasing vulnerability of the gut to pathogenic bacteria. Furthermore, chronic, low‐grade inflammation can have lasting effects on the gut, but the effects of acute inflammation on gut communities have not been thoroughly assessed. In this study, we first investigated whether a single modest dose of LPS administered to adult male and female Siberian hamsters (Phodopus sungorus) activated the immune system by measuring levels of circulating cortisol and the proinflammatory cytokine TNF‐α in the liver compared with saline‐treated animals. We then investigated whether this same acute dose of LPS altered the microbiome 48 h after treatment. We found that, although LPS increased cortisol and liver cytokine levels, and produced changes in food intake and body mass in both sexes, immunological changes were independent of gut dysbiosis 48 h after LPS injection. These data suggest that an acute immune activation may not be capable of altering the gut microbiome in healthy individuals. It is likely, however, that this type of immune challenge may have other physiological impacts on the guts vulnerability, and future studies will investigate these relationships further.

Early-life sickness may predispose Siberian hamsters to behavioral changes following alterations of the gut microbiome in adulthood

Although it is well-established that the immune system plays an important role in the development of physiology and behavior, the gut microbiome has recently become of interest in the study of developmental origins of behavior. Studies suggest that the effects of early-life immune activation may not occur until a secondary stressor is introduced, though the precise nature and timing of the stressor may be critical in the response. Further, recent work suggests that the microbiome and the immune system develop in parallel, and therefore any perturbations to one of these systems early in life will likely affect the other. Here, we sought to determine whether early-life activation of the immune system had long-term consequences on how the gut microbiome responds to antibiotic treatment in adulthood and whether those changes influence adult same-sex social behavior. In order to test the hypothesis that an early-life immune challenge makes individuals more vulnerable to the effects of antibiotics, we mimicked an early-life infection by injecting pups at postnatal day 3 and 5 with lipopolysaccharide (LPS; cell wall component of gram-negative bacteria) or saline, and subsequently exposed the same animals to antibiotic treatment (known to influence microbial community composition and behavior) or water in adulthood. We tracked physiology across development, and paired males and females with a novel individual of the same age and sex in adulthood to score same-sex behavior (e.g., aggression, investigation, grooming) before antibiotic treatment, immediately following treatment, and after recovery from antibiotics. LPS-treated females exhibited impaired reproductive physiology and function in adulthood (e.g., smaller ovaries and abnormal estrous cycles), and female and male gut microbial communities were strongly affected by antibiotic treatment in adulthood, but only slightly affected by postnatal LPS alone. Interestingly, LPS-treated males exhibited more robust changes in their behavioral response following adult antibiotic treatment, including decreased investigation and increased grooming, suggestive of changes in anxiety-like behaviors. These data suggest that males may be more vulnerable than females to behavioral abnormalities after being predisposed to an immune challenge early in life. Collectively, these results provide novel evidence that some of the sex-specific behavioral consequences of an early-life immune challenge may not transpire until an individual is faced with a secondary challenge, and the context in which an individual is exposed can greatly influence the response.

There's no place like biome: Can helminths restore the body's ecosystem?

Immunity is a proverbial double-edged sword. Whereas a competent immune system is crucial for maintaining a sufficiently high level of disease resistance, excessively heightened or over-reactive immune responses may be equally debilitating, leading to a host of allergies and autoimmune diseases. Of considerable concern to immunologists and non-immunologists alike is the dramatic rise in autoimmune and inflammatory disorders during the past decade. A study conducted by the National Center for Health and Statistics determined that the prevalence of asthma has increased from 2001 to 2010 and in 2012 was the highest in recorded history (Akinbami et al., 2012). Immune hyper-reactivity and dysbiosis have also been linked to autism spectrum disorders, anxiety, and obesity (Clarke et al., 2014). Recently, scientists have proffered the ‘‘biome depletion theory” to help make sense of these startling trends. According to this theory, marked increases in autoimmune and inflammatory diseases may be the result of a loss of critical components of our biome (e.g., microbes and multicellular organisms) that normally interact with our immune system. The loss of these microbial communities within modern societies can fuel potentially pathological immune over-reactions (Parker and Ollerton, 2013). The paper by Williamson and colleagues (Williamson et al., 2016) in this issue of Brain, Behavior, and Immunity tests the idea that the loss of biome complexity contributes to hyperactive immune responses. Previous work by this group has demonstrated that neonatal rats infected with the gram negative bacterium Escherichia coli exhibited elevated brain cytokine levels and substantially impaired memory when subsequently exposed to a secondary immune challenge (i.e., lipopolysaccharide) in adulthood (Bilbo et al., 2005; Williamson et al., 2011). The results, while exciting, were also puzzling; why would the immune system have evolved to essentially overreact to exposures that surely must occur frequently in nature? The researchers speculated that perhaps the presence of commensal organisms, particularly gut worms, normally present in the wild (but lacking in most modern, aseptic laboratory rodent colonies), keep immune responses in check. Motivated by this intriguing idea, these researchers tested the effects of prenatal helminth colonization on brain cytokine expression, microglial sensitization, and cognitive dysfunction in response to postnatal bacterial infection in rats (Williamson et al., 2016). Perhaps surprisingly, the results of their study showed that prenatal colonization influenced offspring gut microbiome and immune sensitivity. Critically, helminth exposure early in life prevented the exaggerated brain cytokine response they had previously reported following bacterial infection in adulthood and prevented memory impairment (Williamson et al., 2011). In other words, the presence of the helminths actually buffered the developing immune system of rats against hyper-responsiveness to subsequent immune challenges in adulthood. While the exact relationships between helminths, the brain, and cognition have yet to be fully explored, these findings are exciting as they suggest that ‘‘biome restoration” can alter the immune system and microbial community in important ways that buffer against the adverse effects of neonatal infection. They also suggest the importance of considering inflammation within evolutionary and ecological contexts (Demas and Carlton, 2015). While there is no denying that excessive or unabated inflammation can be detrimental, inflammatory responses more commonly serve to clear individuals of potentially life-threatening infections (Bilbo and Schwarz, 2012). Within this context, the findings of Williamson and colleagues suggest that this ‘‘hyper-reactive” immune response, rather than being maladaptive, is likely an adaptive response to living in an evolutionarily unclean world. Under the artificially aseptic conditions found in most modern laboratories, however, this hyperactivity is most often detrimental to an animal’s health. By creating these aseptic conditions, we are unable to investigate the natural relationships that exist among the different members of the biome. While these findings point to potential opportunities for therapeutic interventions, including the possibility of helminth colonization used to treat conditions like allergies, inflammatory bowel disease, and autism, they have equally important implications for animal welfare and husbandry (Wammes et al., 2014). Quite simply, they provide compelling support for the idea, long appreciated by farmers and their families (but perhaps less so by animal care and use committees) that there may indeed be such a thing as ‘‘too clean.” Collectively, the results from this study clearly demonstrate that organisms, including ourselves, do not exist in a microbial vacuum. Rather, we evolved within complex biomes co-inhabited by a wide range of ‘‘neighbors” (both friend and foe alike),