Ectotherms have peculiar relationships with microorganisms. For instance, bacteria are recovered from the blood and internal organs of healthy teleosts. However, the presence of microbial communities in the healthy teleost brain has not been proposed. Here, we report a living bacterial community in the brain of healthy salmonids with bacterial loads comparable to those of the spleen and 1000-fold lower than in the gut. Brain bacterial communities share >50% of their diversity with gut and blood bacterial communities. Using culturomics, we obtained 54 bacterial isolates from the brains of healthy trout. Comparative genomics suggests that brain bacteria may have adaptations for niche colonization and polyamine biosynthesis. In a natural system, Chinook salmon brain microbiomes shift from juveniles to reproductively mature adults. Our study redefines the physiological relationships between the brain and bacteria in teleosts. This symbiosis may endow salmonids with a direct mechanism to sense and respond to environmental microbes.

Zebrafish (Danio rerio) is now the second most used animal model in biomedical research. As with other vertebrate models, underlying diseases and infections often impact research. Beyond mortality and morbidity, these conditions can compromise research end points by producing nonprotocol induced variation within experiments. Pseudoloma neurophilia, a microsporidium that targets the central nervous system, is the most frequently diagnosed pathogen in zebrafish facilities. The parasite undergoes direct, horizontal transmission within populations, and is also maternally transmitted with spores in ovarian fluid and occasionally within eggs. This transmission explains the wide distribution among research laboratories as new lines are generally introduced as embryos. The infection is chronic, and fish apparently never recover following the initial infection. However, most fish do not exhibit outward clinical signs. Histologically, the parasite occurs as aggregates of spores throughout the midbrain and spinal cord and extends to nerve roots. It often elicits meninxitis, myositis, and myodegeneration when it infects the muscle. There are currently no described therapies for the parasite, thus the infection is best avoided by screening with PCR-based tests and removal of infected fish from a facility. Examples of research impacts include reduced fecundity, behavioral changes, transcriptome alterations, and autofluorescent lesions.

Objective Premature mortality of adult female Chinook Salmon Oncorhynchus tshawytscha is a major barrier to population recovery. The Willamette River basin, Oregon, typifies the problems that are faced by fishery managers in the Pacific Northwest (USA). Adult salmon are trapped and transported upstream of dams to access historical spawning grounds, but annual rates of prespawn mortality (PSM) are high (often >40%) and may limit the recovery of natural populations. The purpose of this study was to identify potential factors related to PSM of female Chinook Salmon that are outplanted above dams and incorporate them into a modeling framework to facilitate adaptive management of outplanting operations. Methods We evaluated PSM in Fall Creek of the Willamette River basin prior to transport facility improvements in summer and fall of 2010-2017 and postimprovement during 2020-2021. We estimated PSM and conducted exploratory analyses to identify possible nontransport sources of stress that may contribute to the observed high PSM rates. Candidate factors included long-term elevated temperature exposure, elevated temperature exposure below the trap, total number of outplanted fish, and monthly human disturbance of outplanted fish. We then developed and fit three models, each representing a hypothesis of a factor influencing PSM, incorporated them into a single alternative decision model, and conducted sensitivity analyses. Result Prespawn mortality averaged 0.66 (ranging from 0.37 to 0.94) over the study period. According to the simulation results, the top two management actions were to exclude human activities-swimming and fishing-from Fall Creek in July and August. Conclusion Expected PSM rates were predicted to be 0.38 when human activity was excluded in July and 0.37 for August. Sensitivity analyses indicated that the most influential decision model component was the choice of the alternative model.

Bighorn sheep (Ovis canadensis) are herbivorous ungulates that live in forage-poor areas of the American west. The trace minerals that herbivores derive from forage are important for immune function. Therefore, identifying trace minerals that affect immune function in bighorn sheep could provide important insights into disease susceptibility and population health in threatened populations. We sought to determine whether trace mineral composition in blood or plasma correlates to survival and determine whether immunologic parameters correlate with any trace minerals that affect survival. We used data collected from 2016 to 2018 as part of a large study on bighorn sheep in southeastern Oregon and northern Nevada, US. We measured the survival of 135 bighorn sheep during the 8-mo monitoring period, including general metrics of immune function and trace mineral levels. We found that animals with higher selenium had improved survival over the monitoring period, with higher peripheral blood mononuclear cell activity (lymphocytes and monocytes) and lower bacterial killing ability in an in vitro assay. This suggests that bighorn sheep may have altered immune function when selenium levels are low, making them more likely to die during the 8-mo monitoring period. Future work should consider whether habitat management strategies that increase selenium intake might improve disease resistance and survival in bighorn sheep in selenium-poor areas.

Objective: Dams and reservoirs can alter juvenile growth and survival of migratory salmonids through several physical and biological mechanisms. Juvenile Chinook Salmon Oncorhynchus tshawytscha that are produced upstream of large hydropower dams may have associated passage mortality, but the reservoirs created by these dams can support rapid growth. Characterizing the biotic drivers of growth and mortality in reservoirs may aid in understanding the cumulative effects of river impoundments on migratory salmonid populations. The purpose of this study was to understand how reservoirs facilitate rapid growth in juvenile Chinook Salmon. Methods: We analyzed stomach contents to determine diet composition throughout the summer and fall. We also recorded prevalence of the parasitic nematode Philonema sp. in the coeloms of fish. Result: We found that juvenile Chinook Salmon frequently consumed young-of-year centrarchids, which likely contributed to rapid growth. Piscivory was highest from July through October and decreased with surface temperature from November through December. Correspondingly, zooplankton and arthropod consumption increased in November and December. Prevalence of visible Philonema sp. infections in the coelom was high (34.6%), negatively associated with time, and nonlinearly associated with fork length. Conclusion: These findings reveal unique diet patterns and suggest potential parasite-associated mortality in reservoir-rearing Chinook Salmon, but more detailed studies across a longer time scale are needed to robustly assess the population-level effects of this parasite.

In species where offspring survival is highly variable relative to adult survival, such as bighorn sheep (Ovis canadensis), physiological indicators of maternal investment could clarify the functional mechanisms of life history trade-offs and serve as important predictors of population dynamics. From a management perspective, simple predictors of juvenile survival measured non-lethally from maternal samples could aid in identifying at-risk populations or individuals before significant mortality occurs. Blood biochemical parameters can offer low-cost insights into animal health and physiology, therefore we sought to develop a simple biochemical predictor of juvenile survival based on maternal blood samples. We measured biochemical indicators of energy balance in adult bighorn sheep at a single time point in January or February, and then monitored survival through August of the same year to assess how those measures related to survival of individual adults and their juvenile offspring. Juvenile survival was lower over the subsequent spring and summer when maternal adult serum beta-hydroxybutyric acid (β-HBA) concentration was high, indicating a negative energy balance in the mothers. However, serum β-HBA did not correlate with adult survival over the same period. Our findings suggest that even when maternal body condition is high, short-term caloric deficit may be sufficient trigger to decrease investment in offspring survival. This mechanism could protect adult females from investing heavily in juvenile survival when resources become too limited to support population growth. Our study suggests that β-HBA could be a powerful monitoring tool for bighorn sheep and other threatened ruminant populations under resource limitation.

Wildlife and natural resource institutions play key roles in invasive species monitoring and management.Paradoxically, the extensive fieldwork undertaken by these institutions and their partners may result in the inadvertent movement and spread of invasive species within and between sensitive ecosystems.In this work, we consider the potential effects of internal guidelines and policies designed to prevent the spread of invasive species by the field activities of management institutions and their partners.Such policies could be high-leverage tools for invasive species prevention, however, as large, complex organizations seek to implement policies to limit the spread of invasive species, it may be challenging to accommodate the wide diversity of potential invasion pathways and mitigation efforts facilitated via the programs, activities, and ecosystems they manage.Prevention policies may also be met with resistance due to the costs of implementation unless concrete benefits can be demonstrated.Assessing and communicating the effects of prevention policies could motivate improved implementation and adherence by institutional units and partners and could help inform adaptive policy changes.However, assessing the effectiveness of invasive species prevention presents a unique set of challenges, including incomplete data on invasive species distribution and pathways, that make it difficult to measure the effects of prevention efforts.In this work, we present a conceptual framework for evaluating institutional policies for invasive species prevention.We describe a flexible, multifaceted approach that considers policy implementation and adherence as well as ecological outcomes.We discuss potential application of this framework using a policy recently implemented by the Pacific Region of the U.S. Fish and Wildlife Service to prevent the introduction and spread of invasive species by service personnel and partners during field activities as a case study.

Ectotherms have long been known to have peculiar relationships with microorganisms. For instance, bacteria can be recovered from blood and internal organs of healthy teleost fish. However, until now, the presence of a microbial community in the healthy teleost brain has not been proposed. Here we report a living bacterial community in the brain of healthy salmonids. Brain bacterial loads in salmonids are comparable to those found in the spleen and 1000-fold lower than in the gut. Brain bacterial communities share >50% of their diversity with gut and blood bacterial communities. Using culturomics, we obtained 54 bacterial isolates from the brain of healthy rainbow trout. Comparative genomics uncovered unique niche adaptations associated with brain colonization and polyamine biosynthesis. In a natural system, salmonid brain microbiomes shift with the host life cycle, becoming dysbiotic in reproductively mature Chinook salmon, a species that undergoes reproductive death. Our study redefines the relationship between the teleost brain and bacterial microbiomes under physiological conditions. We posit that this symbiosis may endow salmonids with a direct mechanism to sense and respond to environmental microbes.One-Sentence Summary Salmonids have a brain-adapted, resident bacterial community### Competing Interest StatementThe authors have declared no competing interest.

Pacific salmon experience prolonged elevation in corticosteroid hormones during important life history events including migration, reproduction, and senescence. These periods of elevated corticosteroids correspond with changes to immunity and energy metabolism; therefore, fish may be particularly vulnerable to mortality at these times. Recent studies found that stress-induced cortisol release associated with microbial community shifts in salmonids, raising the question of how longer-term corticosteroid dynamics that accompany life history transitions affect salmonid microbiomes. In this work, we experimentally evaluated the relationships between gut microbiome composition, chronically elevated corticosteroids, and mortality in juvenile Chinook salmon ( Oncorhynchus tshawytscha ). We found that treatment with slow-release implants of the corticosteroids cortisol or dexamethasone resulted in changes to the gut microbiome. Morbidity was also associated with microbiome composition, suggesting that the gut microbiome reflects individual differences in susceptibility to opportunistic pathogens. Additionally, we analyzed a small number of samples from adult fish at various stages of senescence. Results from these samples suggest that microbiome composition associated with gut integrity, and that the microbial communities of corticosteroid treated juveniles shift in composition toward those of senescent adults. Overall, findings from this work indicate that the gut microbiome correlates with mortality risk during periods of chronic corticosteroid elevation.

A novel Enterocytozoon infection was identified in the intestines of sexually mature Chinook salmon. While microsporidian parasites are common across a diverse range of animal hosts, this novel species is remarkable because it demonstrates biological, pathological, and genetic similarity with Enterocytozoon bieneusi, the most common causative agent of microsporidiosis in AIDS patients. There are similarities in the immune and endocrine processes of sexually mature Pacific salmon and immunocompromised humans, suggesting possible common mechanisms of susceptibility in these two highly divergent host species. The discovery of Enterocytozoon schreckii n. sp. contributes to clarifying the phylogenetic relationships within family Enterocytozoonidae. The phylogenetic and morphological features of this species support the redescription of Enterocytozoon to include Enterospora as a junior synonym. Furthermore, the discovery of this novel parasite may have important implications for conservation, as it could be a sentinel of immune suppression, disease, and prespawning mortality in threatened populations of salmonids. IMPORTANCE In this work, we describe a new microsporidian species that infects the enterocytes of Chinook salmon. This novel pathogen is closely related to Enterocytozoon bieneusi, an opportunistic pathogen commonly found in AIDS patients and other severely immunocompromised humans. The discovery of this novel pathogen is of interest because it has only been found in sexually mature Chinook salmon, which have compromised immune systems due to the stresses of migration and maturation and which share similar pathological features with immunocompromised and senescent humans. The discovery of this novel pathogen could lead to new insights regarding how microsporidiosis relates to immunosuppression across animal hosts.

In recent years, emerging sequencing technologies and computational tools have driven a tidal wave of research on host-associated microbiomes, particularly the gut microbiome. These studies demonstrate numerous connections between the gut microbiome and vital host functions, primarily in humans, model organisms, and domestic animals. As the adaptive importance of the gut microbiome becomes clearer, interest in studying the gut microbiomes of wild populations has increased, in part due to the potential for discovering conservation applications. The study of wildlife gut microbiomes holds many new challenges and opportunities due to the complex genetic, spatial, and environmental structure of wild host populations, and the potential for these factors to interact with the microbiome. The emerging picture of adaptive coevolution in host-microbiome relationships highlights the importance of understanding microbiome variation in the context of host population genetics and landscape heterogeneity across a wide range of host populations. We propose a conceptual framework for understanding wildlife gut microbiomes in relation to landscape variables and host population genetics, including the potential of approaches derived from landscape genetics. We use this framework to review current research, synthesize important trends, highlight implications for conservation, and recommend future directions for research. Specifically, we focus on how spatial structure and environmental variation interact with host population genetics and microbiome variation in natural populations, and what we can learn from how these patterns of covariation differ depending on host ecological and evolutionary traits.

Measuring inflammatory markers is critical to evaluating both recent infection status and overall human and animal health; however, there are relatively few techniques that do not require specialized equipment or personnel for detecting inflammation among wildlife. Such techniques are useful in that they help determine individual and population-level inflammatory status without the infrastructure and reagents that many more-specific assays require. One such technique, known as the erythrocyte sedimentation rate (ESR), is a measure of how quickly erythrocytes (red blood cells) settle in serum, with a faster rate indicating a general, underlying inflammatory process is occurring. The technique is simple, inexpensive, and can be performed in the field without specialized equipment. We took advantage of a population of African buffalo (Syncerus caffer), well studied from June 2014 to May 2017, to understand the utility of ESR in an important wildlife species. When ESR was compared with other markers of immunity in African buffalo, it correlated to known measures of inflammation. We found that a faster ESR was significantly positively correlated with increased total globulin levels and significantly negatively correlated with increased red blood cell count and albumin levels. We then evaluated if ESR correlated to the incidence of five respiratory pathogens and infection with two tick-borne pathogens in African buffalo. Our results suggest that elevated ESR is associated with the incidence of bovine viral diarrhea virus infection, parainfluenza virus, and Mannheimia haemolytica infections as well as concurrent Anaplasma marginale and Anaplasma centrale coinfection. These findings suggest that ESR is a useful field test as an inflammatory marker in individuals and herds, helping us better monitor overall health status in wild populations.

Arctic species are likely to experience rapid shifts in prey availability under climate change, which may alter their exposure to microbes and parasites. Here, we describe fecal bacterial and macroparasite communities and assess correlations with diet trophic level in Pacific walruses harvested during subsistence hunts by members of the Native Villages of Gambell and Savoonga on St Lawrence Island, Alaska. Fecal bacterial communities were dominated by relatively few taxa, mostly belonging to phyla Fusobacteriota and Firmicutes. Members of parasite-associated phyla Nematoda, Acanthocephala and Platyhelminthes were prevalent in our study population. We hypothesized that high versus low prey trophic level (e.g. fish versus bivalves) would result in different gut bacterial and macroparasite communities. We found that bacterial community structure correlated to diet, with nine clades enriched in walruses consuming higher-trophic-level prey. While no parasite compositional differences were found at the phylum level, the cestode genus Diphyllobothrium was more prevalent and abundant in walruses consuming higher-trophic-level prey, probably because fish are the intermediate hosts for this genus. This study suggests that diet is important for structuring both parasite and microbial communities of this culturally and ecologically important species, with potential implications for population health under climate change.

Taxonomic classifications for all eukaryotic OTUs identified from 18S sequence data.

There have been several significant new findings regarding Microsporidia of fishes over the last decade. Here we provide an update on new taxa, new hosts and new diseases in captive and wild fishes since 2013. The importance of microsporidiosis continues to increase with the rapid growth of finfish aquaculture and the dramatic increase in the use of zebrafish as a model in biomedical research. In addition to reviewing new taxa and microsporidian diseases, we include discussions on advances with diagnostic methods, impacts of microsporidia on fish beyond morbidity and mortality, novel findings with transmission and invertebrate hosts, and a summary of the phylogenetics of fish microsporidia.

In semelparous Pacific salmon, increased cortisol levels accompany sexual maturation and may be related to the rapid senescence and death that occur after spawning. In fish with extremely high cortisol, pre-spawning mortality is more likely. This may be because elevated cortisol is accompanied by energy depletion and reduces the immune capacity of sexually maturing individuals, thus increasing their susceptibility to parasites and pathogens. Several studies have measured cortisol levels in Pacific salmon during the last few weeks prior to spawning, but there is a lack of information regarding longer-term changes in resting and stressed cortisol levels of migrating adult Pacific salmon. A better understanding of the scope of the cortisol response during sexual maturation could contribute to understanding the extremely high pre-spawning mortality experienced by some threatened populations of Pacific salmon. The objective of this study was to determine the scope of the cortisol stress response in spring Chinook salmon as well as the dynamics of resting cortisol during maturation. We conducted a laboratory experiment in which plasma cortisol in adult Chinook salmon was measured before and after application of a standardized stressor applied at monthly intervals during the last three months prior to spawning. We found that resting and stressed cortisol levels increased during the last three months prior to spawning. We also found that sexually maturing Chinook salmon are able to mount a cortisol response to acute stressors when resting levels are elevated during maturation. Additionally, the effects of stress and time on cortisol dynamics differed between males and females as well as between individuals, which has implications for population resilience to anthropogenic stressors in wild populations.

Studies in humans and laboratory animals link stable gut microbiome "enterotypes" with long-term diet and host health. Understanding how this paradigm manifests in wild herbivores could provide a mechanistic explanation of the relationships between microbiome dynamics, changes in dietary resources, and outcomes for host health. We identify two putative enterotypes in the African buffalo gut microbiome. The enterotype prevalent under resource-abundant dietary regimes, regardless of environmental conditions, has high richness, low between- and within-host beta diversity, and enrichment of genus Ruminococcaceae-UCG-005. The second enterotype, prevalent under restricted dietary conditions, has reduced richness, elevated beta diversity, and enrichment of genus Solibacillus. Population-level gamma diversity is maintained during resource restriction by increased beta diversity between individuals, suggesting a mechanism for population-level microbiome resilience. We identify three pathogens associated with microbiome variation depending on host diet, indicating that nutritional background may impact microbiome-pathogen dynamics. Overall, this study reveals diet-driven enterotype plasticity, illustrates ecological processes that maintain microbiome diversity, and identifies potential associations between diet, enterotype, and disease.

Objectives Methylmercury metabolism was investigated in Pacific walruses (Odobenus rosmarus divergens) from St. Lawrence Island, Alaska, United States. Methods Total mercury and methylmercury concentrations were measured in fecal samples and paired colon samples (n = 16 walruses). Gut microbiota composition and diversity were determined using 16S rRNA gene sequencing. Associations between fecal and colon mercury and the 24 most prevalent gut microbiota taxa were investigated using linear models. Results In fecal samples, the median values for total mercury, methylmercury, and %methylmercury (of total mercury) were 200 ng/g, 4.7 ng/g, and 2.5%, respectively, while in colon samples, the median values for the same parameters were 28 ng/g, 7.8 ng/g, and 26%, respectively. In fecal samples, methylmercury was negatively correlated with one Bacteroides genus, while members of the Oscillospirales order were positively correlated with both methylmercury and %methylmercury (of total mercury). In colon samples, %methylmercury (of total mercury) was negatively correlated with members of two genera, Romboutsia and Paeniclostridium. Conclusions Median %methylmercury (of total mercury) was 10 times higher in the colon compared to the fecal samples, suggesting that methylmercury was able to pass through the colon into systemic circulation. Fecal total mercury and/or methylmercury concentrations in walruses were comparable to some human studies despite differences in seafood consumption rates, suggesting that walruses excreted less mercury. There are no members (at this time) of the Oscillospirales order which are known to contain the genes to methylate mercury, suggesting the source of methylmercury in the gut was from diet and not in vivo methylation.

Supplemental feeding of wildlife is a common practice often undertaken for recreational or management purposes, but it may have unintended consequences for animal health. Understanding cryptic effects of diet supplementation on the gut microbiomes of wild mammals is important to inform conservation and management strategies. Multiple laboratory studies have demonstrated the importance of the gut microbiome for extracting and synthesizing nutrients, modulating host immunity, and many other vital host functions, but these relationships can be disrupted by dietary perturbation. The well-described interplay between diet, the microbiome, and host health in laboratory and human systems highlights the need to understand the consequences of supplemental feeding on the microbiomes of free-ranging animal populations. This study describes changes to the gut microbiomes of wild elk under different supplemental feeding regimes. We demonstrated significant cross-sectional variation between elk at different feeding locations and identified several relatively low-abundance bacterial genera that differed between fed versus unfed groups. In addition, we followed four of these populations through mid-season changes in supplemental feeding regimes and demonstrated a significant shift in microbiome composition in a single population that changed from natural forage to supplementation with alfalfa pellets. Some of the taxonomic shifts in this population mirrored changes associated with ruminal acidosis in domestic livestock. We discerned no significant changes in the population that shifted from natural forage to hay supplementation, or in the populations that changed from one type of hay to another. Our results suggest that supplementation with alfalfa pellets alters the native gut microbiome of elk, with potential implications for population health.