Zoos as Sentinels? A Meta-Analysis of Seroprevalence of Terrestrial Mammalian Viruses in Zoos.

The One Health framework links animal, human, and environmental health, and focuses on emerging zoonotic pathogens. Understanding the interface between wildlife and human activity is critical due to the unpredictable nature of spillover of zoonotic pathogens from animals to humans. Zoos are important partners in One Health because of their contributions to education, conservation, and animal health monitoring. In addition, the housing of wildlife in captive and semi-natural settings makes zoos, especially relevant for detecting animal-related pathogens. A first step to determine the utility of zoos in contributing to pathogen surveillance is to survey the peer-reviewed literature. We, therefore, retrieved data from the previous 20 years and performed a meta-analysis to determine global patterns of viral seroprevalence in mammals housed in zoo collections from peer-reviewed literature. We analysed 50 articles, representing a total of 11,300 terrestrial mammals. Increased prevalence was found in viruses strictly targeting specific host taxonomy, especially in viruses transmitted through direct contact. Potentially complex patterns with geography were also identified, despite uneven sampling. This research highlights the role zoos could play in public health and encourages future standardized epidemiological surveillance of zoological collections.

Tick infestation effects on haemoglobin levels of deer mice (Peromyscus maniculatus).

Deer mice (Peromyscus maniculatus) are hosts to ixodid ticks as well as the associated tick-borne pathogens they can spread. As the ranges of black-legged ticks (Ixodes scapularis) and American dog ticks (Dermacentor variabilis) expand northwards, naïve host populations of deer mice are likely to become infested by ticks and experience the physiological effects that ticks can have on them via blood-feeding. The prevalence of these haematophagous ticks can affect the haemoglobin levels of the mice they infest. Haemoglobin levels were compared and analysed in deer mice populations at three different sites with varying tick exposure. These results suggested that without confounding effects, the abundance of black-legged and American dog ticks on individual mice had a significant negative effect on the hosts' haemoglobin levels, but only in an area with high tick infestation. This was seen across the average haemoglobin levels between populations, where there was a significant difference between the source population with the longest established tick populations and the source population where neither black-legged nor American dog ticks were prevalent. As the ticks' ranges expand and they become more abundant, it is important to understand how their prevalence and intensity can alter host physiology, potentially affecting their own range expansion and the spread of the diseases they may carry.

Effects of ticks on community assemblages of ectoparasites in deer mice.

Ectoparasites are fundamental to ecosystems, playing a key role in trophic regulation. Fleas, mites, and ticks are common hematophagous ectoparasites that infest shared mammalian hosts. One common host in Ontario, Canada, is the deer mouse (Peromyscus maniculatus). As the climate warms and the geographic ranges of blacklegged ticks (Ixodes scapularis) and American dog ticks (Dermacentor variabilis) expand, their introduction to new ecosystems may alter current ectoparasite communities. At three different sites where exposure to ticks varied (both in terms of tick diversity and abundance), ectoparasite community structures found on deer mouse hosts were examined, focusing on species co-occurrences and habitat partitioning on the host. We predicted that when tick species were prevalent, ticks would dominate the micro-habitat attachment sites often inhabited by other parasites, thereby significantly altering parasite community structure. Our results suggest that blacklegged ticks and American dog ticks could have a positive association with each other, but a negative or random association with other ectoparasite species, even when they do not occupy the same attachment site. Sampling site played a significant role in community assemblages as well, possibly due to the differences in tick exposure. As the ticks' ranges expand and they become more abundant, it is important to understand how their prevalence can potentially alter the dynamics in an ectoparasite community, affecting the transmission of pathogens that may spread within an ecosystem, from one host to another.

Corticosterone response by Peromyscus mice to parasites, reproductive season, and age.

A common response to parasite infestations is increased production of glucocorticoid hormones that regulate immune function. We examined relationships between ectoparasite infestations and fecal corticosterone metabolites (FCM) in deer mice (Peromyscus maniculatus). Furthermore, we experimentally removed fleas to determine if reductions in ectoparasites affected FCM production. Individuals were assigned to control (no flea removal) or treatment (anti-flea application, physical combing) groups and individuals were recaptured to assess changes in FCM concentrations. There was a significant and negative effect of number of anti-flea treatment applications on FCM concentrations of deer mice. However, models including host biology traits and environmental predictors had a better model fit compared to models containing ectoparasite predictors. In particular, there was a significant relationship of deer mouse FCM with date and host age, where glucocorticoid production decreased towards the end of the breeding season and increased with age. Overall, adverse events associated with reproduction and age class, rather than ectoparasites, may be more important to variation in glucocorticoids of deer mice.

Functional genetic diversity of domestic and wild American mink (Neovison vison).

The release of domestic organisms to the wild threatens biodiversity because the introduction of domestic genes through interbreeding can negatively impact wild conspecifics via outbreeding depression. In North America, farmed American mink (Neovison vison) frequently escape captivity, yet the impact of these events on functional genetic diversity of wild mink populations is unclear. We characterized domestic and wild mink in Ontario at 17 trinucleotide microsatellites located in functional genes thought to be associated with traits affected by domestication. We found low functional genetic diversity in both mink types, as only four of 17 genes were variable, yet allele frequencies varied widely between captive and wild populations. To determine whether allele frequencies of wild populations were affected by geographic location, we performed redundancy analysis and spatial analysis of principal components on three polymorphic loci (AR, ATN1 and IGF-1). We found evidence to suggest domestic release events are affecting the functional genetic diversity of wild mink, as sPCA showed clear distinctions between wild individuals near mink farms and those located in areas without mink farms. This is further substantiated through RDA, where spatial location was associated with genetic variation of AR, ATN1 and IGF1.

Corrigendum to "Parasite species co-occurrence patterns on Peromyscus: Joint species distribution modelling" [IJP: Parasites Wildlife 12 (2020) 199-206].

[This corrects the article DOI: 10.1016/j.ijppaw.2020.04.011.].

Population genomic evidence that human and animal infections in Africa come from the same populations of Dracunculus medinensis.

BACKGROUND: Guinea worm-Dracunculus medinensis-was historically one of the major parasites of humans and has been known since antiquity. Now, Guinea worm is on the brink of eradication, as efforts to interrupt transmission have reduced the annual burden of disease from millions of infections per year in the 1980s to only 54 human cases reported globally in 2019. Despite the enormous success of eradication efforts to date, one complication has arisen. Over the last few years, hundreds of dogs have been found infected with this previously apparently anthroponotic parasite, almost all in Chad. Moreover, the relative numbers of infections in humans and dogs suggests that dogs are currently the principal reservoir on infection and key to maintaining transmission in that country. PRINCIPAL FINDINGS: In an effort to shed light on this peculiar epidemiology of Guinea worm in Chad, we have sequenced and compared the genomes of worms from dog, human and other animal infections. Confirming previous work with other molecular markers, we show that all of these worms are D. medinensis, and that the same population of worms are causing both infections, can confirm the suspected transmission between host species and detect signs of a population bottleneck due to the eradication efforts. The diversity of worms in Chad appears to exclude the possibility that there were no, or very few, worms present in the country during a 10-year absence of reported cases. CONCLUSIONS: This work reinforces the importance of adequate surveillance of both human and dog populations in the Guinea worm eradication campaign and suggests that control programs aiming to interrupt disease transmission should stay aware of the possible emergence of unusual epidemiology as pathogens approach elimination.

Parasite species co-occurrence patterns on Peromyscus: Joint species distribution modelling.

Hosts are often infested by multiple parasite species, but it is often unclear whether patterns of parasite co-occurrence are driven by parasite habitat requirements or parasite species interactions. Using data on infestation patterns of ectoparasitic arthropods (fleas, trombiculid mites, cuterebrid botflies) from deer mice (Peromyscus maniculatus), we analyzed species associations using joint species distribution modelling. We also experimentally removed a flea (Orchopeas leucopus) from a subset of deer mice to examine the effect on other common ectoparasite species. We found that the mite (Neotrombicula microti) and botfly (Cuterebra sp.) had a negative relationship that is likely a true biotic species interaction. The flea had a negative association with the mite and a positive association with the botfly species, both of which appeared to be influenced by host traits or parasite life-history traits. Furthermore, experimental removal of the flea did not have a significant effect on ectoparasite prevalence of another species. Overall, these findings suggest that complex parasite species associations can be present among multiple parasite taxa, and that aggregation is not always the rule for ectoparasite communities of small mammals.

Effects of captivity, diet, and relocation on the gut bacterial communities of white-footed mice.

Microbes can have important impacts on their host's survival. Captive breeding programs for endangered species include periods of captivity that can ultimately have an impact on reintroduction success. No study to date has investigated the impacts of captive diet on the gut microbiota during the relocation process of generalist species. This study simulated a captive breeding program with white-footed mice (Peromyscus leucopus) to describe the variability in gut microbial community structure and composition during captivity and relocation in their natural habitat, and compared it to wild individuals. Mice born in captivity were fed two different diets, a control with dry standardized pellets and a treatment with nonprocessed components that reflect a version of their wild diet that could be provided in captivity. The mice from the two groups were then relocated to their natural habitat. Relocated mice that had the treatment diet had more phylotypes in common with the wild-host microbiota than mice under the control diet or mice kept in captivity. These results have broad implications for our understanding of microbial community dynamics and the effects of captivity on reintroduced animals, including the potential impact on the survival of endangered species. This study demonstrates that ex situ conservation actions should consider a more holistic perspective of an animal's biology including its microbes.

Selective whole genome amplification and sequencing of Coxiella burnetii directly from environmental samples.

Whole genome sequencing (WGS) is a widely available, inexpensive means of providing a wealth of information about an organism's diversity and evolution. However, WGS for many pathogenic bacteria remain limited because they are difficult, slow and/or dangerous to culture. To avoid culturing, metagenomic sequencing can be performed directly on samples, but the sequencing effort required to characterize low frequency organisms can be expensive. Recently developed methods for selective whole genome amplification (SWGA) can enrich target DNA to provide efficient sequencing. We amplified Coxiella burnetii (a bacterial select agent and human/livestock pathogen) from 3 three environmental samples that were overwhelmed with host DNA. The 68- to 147-fold enrichment of the bacterial sequences provided enough genome coverage for SNP analyses and phylogenetic placement. SWGA is a valuable tool for the study of difficult-to-culture organisms and has the potential to facilitate high-throughput population characterizations as well as targeted epidemiological or forensic investigations.

Effects of the captive and wild environment on diversity of the gut microbiome of deer mice (Peromyscus maniculatus).

Vertebrate gastrointestinal tracts have co-existed with microbes over millennia. These microbial communities provide their host with numerous benefits. However, the extent to which different environmental factors contribute to the assemblage of gut microbial communities is not fully understood. The purpose of this study was to determine how the external environment influences the development of gut microbiome communities (GMCs). Faecal samples were collected from deer mice (Peromyscus maniculatus) born and raised in captivity and the wild at approximately 3-5 weeks of age. Additional samples were collected 2 weeks later, with a subset of individuals being translocated between captive and wild environments. Microbial data were analysed using 16S rRNA next-generation Illumina HiSeq sequencing methods. GMCs of deer mice were more similar between neighbours who shared the same environment, regardless of where an individual was born, demonstrating that GMCs are significantly influenced by the surrounding environment and can rapidly change over time. Mice in natural environments contained more diverse GMCs with higher relative abundances of Ruminoccocaceae, Helicobacteraceae and Lachnospiraceae spp. Future studies should examine the fitness consequences associated with the presence/absence of microbes that are characteristic of GMCs of wild populations to gain a better understanding of environment-microbe-host evolutionary and ecological relationships.

The effects of body region, season and external arsenic application on hair cortisol concentration.

Hair cortisol analysis has been used to quantify hormone levels in circulation in several mammal species. Hair remains stable for decades or centuries, allowing researchers to use archived hair samples to investigate hormone levels that span long time periods. However, several studies have found that intra-individual variability, driven by the body region from which a sample is derived, confounds measurements of systemic glucocorticoid hormone concentrations. In addition, the external application of chemical agents to hair can remove or concentrate molecules of interest. These may preclude the use of samples that have been collected opportunistically and/or those that have been housed in museum collections. Using a captive population of Vancouver Island marmots (Marmota vancouverensis), we found a strong effect of body region on the concentration of cortisol within hair, as well as an effect of season. Using a collection of American mink (Neovison vison) pelts, we found that application of the preservative arsenic in the form of a soap does not cause a significant decrease in cortisol. The marmot results suggest that intra-individual variability is not stable through time. The reason for these seasonal effects is not clear and further study is necessary. Researchers using samples from an unknown body region should exercise caution in interpreting their results. The mink results suggest that samples held in museum collections can be used to quantify cortisol, even when arsenic preservation is suspected.

Enhanced access to anthropogenic food waste is related to hyperglycemia in raccoons (Procyon lotor).

Urban landscapes have well-known effects on wildlife populations. Many species of urban wildlife feed on anthropogenic food wastes, and little is known regarding the sub-lethal physiological consequences of this novel diet. We use samples from three populations of raccoons to test the hypothesis that access to anthropogenic food waste will lead to elevated body mass, blood glucose and serum leptin. Each population varied in their presumed access to food waste. We found that raccoons from the site with the highest presumed access to food waste were significantly heavier and had significantly higher levels of glycated serum protein (GSP, a marker of elevated blood glucose). In addition, GSP concentration was positively related to body mass. No significant differences in serum leptin were detected, nor was serum leptin related to body mass. Urban diets may have significant physiological consequences for urban wildlife related to glucose metabolism. Further research will be needed to determine the evolutionary consequences of the novel urban diet, and whether adaptation is occurring.

Temporal variation of the microbiome is dependent on body region in a wild mammal (Tamiasciurus hudsonicus).

Microbial communities are increasingly being recognized as important to host health in wild mammals, but how these communities are characterized can have important consequences on the results of these studies. Previous research has explored temporal variation in microbial communities in humans and lab mammals, but few have investigated how microbiomes fluctuate in wild populations and none have examined the temporal dynamics of these fluctuations in different body regions on a wild mammal. Using Illumina MiSeq sequencing of the V3-V4 16S rRNA gene regions, we characterized the buccal and gut microbiomes of wild North American red squirrels, Tamiasciurus hudsonicus, to measure changes in these two microbiomes over short (<2 weeks), medium (2-4 weeks) and long (>1 month) term sampling periods. While we observed short and medium temporal stability in the buccal microbiome, the gut microbiome varied between medium and long-term sampling periods. There was no evidence of intra-individual correlations between buccal and gut microbiome change, suggesting that temporal stability is dependent on the body region and factors affecting microbial stability may be specific to body sites. From these findings, we urge researchers to be cautious in interpreting results from single temporal sampling periods when quantifying characteristic microbiomes in wild mammals.

Stress and the microbiome: linking glucocorticoids to bacterial community dynamics in wild red squirrels.

Bacterial diversity within animals is emerging as an essential component of health, but it is unknown how stress may influence the microbiome. We quantify a proximate link between the oral microbiome and hypothalamic-pituitary-adrenal (HPA) axis activity using faecal glucocorticoid metabolites (FGM) in wild red squirrels (Tamiasciurus hudsonicus). Not only was bacterial diversity lower at higher levels of FGM, but also between capture periods a change in bacterial relative abundance was related to an increase in FGM. These linkages between the HPA axis and microbiome communities represent a powerful capacity for stress to have multi-dimensional effects on health.

Integrating evolution in the management of captive zoo populations.

Both natural animal populations and those in captivity are subject to evolutionary forces. Evolutionary changes to captive populations may be an important, but poorly understood, factor that can affect the sustainability of these populations. The importance of maintaining the evolutionary integrity of zoo populations, especially those that are used for conservation efforts including reintroductions, is critical for the conservation of biodiversity. Here, we propose that a greater appreciation for an evolutionary perspective may offer important insights that can enhance the reproductive success and health for the sustainability of captive populations. We provide four examples and associated strategies that highlight this approach, including minimizing domestication (i.e., genetic adaptation to captivity), integrating natural mating systems into captive breeding protocols, minimizing the effects of translocation on variation in photoperiodism, and understanding the interplay of parasites/pathogens and inflammation. There are a myriad of other issues that may be important for captive populations, and we conclude that these may often be species specific. Nonetheless, an evolutionary perspective may mitigate some of the challenges currently facing captive populations that are important from a conservation perspective, including their sustainability.

Aleutian mink disease virus in striped skunks (Mephitis mephitis): evidence for cross-species spillover.

Aleutian mink disease virus (AMDV) causes a parvovirus infection, initially characterized in American mink (Neovison vison), that may have harmful effects on wild populations of susceptible animals. In North America, where American mink are native, the origin, host range, and prevalence of AMDV in wild species is not clear. We studied striped skunks (Mephitis mephitis) and raccoons (Procyon lotor) to determine whether species sympatric with mink are potential reservoirs in the transmission of AMDV to wild mink and mink farms. Antibodies to AMDV were detected in 41% of skunk serum samples (143/347) and AMDV nucleic acids were detected in 32% (14/40) of skunk spleen samples by PCR, indicating that AMDV exposure and infection were frequent in skunks. We detected no AMDV antibodies in 144 raccoon blood samples. Phylogenetic analysis revealed a newly identified AMDV haplogroup consisting of isolates from Ontario skunks and a free-ranging domestic mink from Ontario. Our findings of frequent AMDV infection in skunks, close genetic similarity between skunk and mink AMDV isolates, and evidence of AMDV transmission from skunks to mink support the hypothesis that skunks may be acting as alternative hosts and reservoirs of AMDV to wild mink through cross-species virus spillover.

Towards improving the ethics of ecological research.

We argue that the ecological research community should develop a plan for improving the ethical consistency and moral robustness of the field. We propose a particular ethics strategy--specifically, an ongoing process of collective ethical reflection that the community of ecological researchers, with the cooperation of applied ethicists and philosophers of biology, can use to address the needs we identify. We suggest a particular set of conceptual (in the form of six core values--freedom, fairness, well being, replacement, reduction, and refinement) and analytic (in the forms of decision theoretic software, 1000Minds) tools that, we argue, collectively have the resources to provide an empirically grounded and conceptually complete foundation for an ethics strategy for ecological research. We illustrate our argument with information gathered from a survey of ecologists conducted at the 2013 meeting of the Canadian Society of Ecology and Evolution.

The ethical dimensions of wildlife disease management in an evolutionary context.

Best practices in wildlife disease management require robust evolutionary ecological research (EER). This means not only basing management decisions on evolutionarily sound reasoning, but also conducting management in a way that actively contributes to the on-going development of that research. Because good management requires good science, and good science is 'good' science (i.e., effective science is often science conducted ethically), good management therefore also requires practices that accord with sound ethical reasoning. To that end, we propose a two-part framework to assist decision makers to identify ethical pitfalls of wildlife disease management. The first part consists of six values - freedom, fairness, well-being, replacement, reduction, and refinement; these values, developed for the ethical evaluation of EER practices, are also well suited for evaluating the ethics of wildlife disease management. The second part consists of a decision tree to help identify the ethically salient dimensions of wildlife disease management and to guide managers toward ethically responsible practices in complex situations. While ethical reasoning cannot be used to deduce from first principles what practices should be undertaken in every given set of circumstances, it can establish parameters that bound what sorts of practices will be acceptable or unacceptable in certain types of scenarios.