Strongyloides Robustus Research Articles

Testing the parasite-mediated competition hypothesis between sympatric northern and southern flying squirrels.

Competition is a driving factor in shaping ecological communities and may act directly or indirectly through apparent competition. We examined a classic example of parasite-mediated competition between northern (Glaucomys sabrinus) and southern flying squirrels (G. volans) via the intestinal nematode, Strongyloides robustus, and tested whether it could act as a species barrier in a flying squirrel hybrid zone. We live-trapped flying squirrels (G. sabrinus and G. volans), grey squirrels (Sciurus carolinensis), red squirrels (Tamiasciurus hudsonicus), and chipmunks (Tamias striatus) from June–September 2019 at 30 woodlots in Ontario, Canada. Fecal samples from squirrels were collected and analyzed for the presence of endoparasite eggs. For each individual, we calculated Scaled Mass Index (SMI) as a measure of body condition to assess the effect of S. robustus on squirrels. We found eggs of S. robustus in all species except chipmunks. Infection with S. robustus did not appear to affect body condition of southern flying squirrels and grey squirrels, but we did find a weak negative effect on northern flying squirrels and red squirrels. Despite a weak asymmetric effect of S. robustus on flying squirrels, we did not find any evidence that parasite-mediated competition could lead to competitive exclusion from woodlots. Furthermore, S. robustus eggs were common in feces of the red squirrel, a species largely sympatric with northern flying squirrel.

Invading parasites: spillover of an alien nematode reduces survival in a native species

It is widely assumed that spillover of alien parasites to native host species severely impacts naïve populations, ultimately conferring a competitive advantage to invading hosts that introduced them. Despite such host-switching events occurring in biological invasions, studies demonstrating the impact of alien macroparasites on native animal hosts are surprisingly few. In Europe, native red squirrels (Sciurus vulgaris) are replaced by introduced North American grey squirrels (S. carolinensis) mainly through resource competition, and, only in the United Kingdom and Ireland, by competition mediated by a viral disease. In Italy such disease is absent, but spillover of an introduced North American nematode (Strongyloides robustus) from grey to red squirrels is known to occur. Here, we used long-term (9 years) capture-mark-recapture and parasitological data of red squirrels in areas co-inhabited by grey squirrels in Northern Italy to investigate the impact of this alien helminth on naïve native squirrels’ body mass, local survival, and reproduction of females. We found no negative effect of the alien parasite on body mass or reproductive success, but intensity of infection by S. robustus reduced survival of both male and female squirrels. Significantly, survival of squirrels co-infected by their native nematode, Trypanoxyuris sciuri, was less affected by S. robustus, suggesting a protective effect of the native helminth against the new infection. Hence, we demonstrate that alien S. robustus spillover adds to the detrimental effects of resource competition and stress induced by grey squirrels, further reducing the fitness of the native species in the presence of the invasive competitor.

Complex relationships between physiological stress and endoparasite infections in natural populations.

Short-term elevation of glucocorticoids (GCs) is one of the major physiological mechanisms by which vertebrates cope with challenging environmental or social factors (stressors). However, when exposure to stressors occurs repeatedly or over a prolonged period of time, animals may experience chronic elevation of GCs, which reduces the immune response efficiency and can lead to higher intensity of parasitic infection. Here, we used invasive gray squirrels Sciurus carolinensis introduced in Northern Italy and their 2 most prevalent gastrointestinal parasites, the nematode Strongyloides robustus and coccidia of the genus Eimeria, as a model to investigate relationships among macroparasite infection and concentrations of fecal glucocorticoid metabolites (FGMs), an integrated measure of circulating GCs. Our results revealed an association of FGMs with infection by St. robustus, but not with coccidia. Individuals with higher FGMs appear to be responsible for the greatest St. robustus egg shedding within gray squirrel populations, thus possibly acting as superspreaders. However, FGMs were negatively associated with adult St. robustus, suggesting that the abundance of adults of this nematode species does not induce elevation in FGMs, but is only affected by it through immune-mediated effects on its fecundity. Finally, the relationship between St. robustus (both eggs and adult parasites) and FGMs was not linear, suggesting that only high levels of physiological stress influence parasite infection. Our findings highlight that the direction and magnitude of the stress–infection relationship may depend not only on the specific host–parasite system, but also on the different life stages of the same parasite.

Spillover of an alien parasite reduces expression of costly behaviour in native host species.

Understanding the effects of invasive alien species (IAS) on native host-parasite relationships is of importance for enhancing ecological theory and IAS management. When IAS and their parasite(s) invade a guild, the effects of interspecific resource competition and/or parasite-mediated competition can alter existing native host-parasite relationships and the dependent biological traits such as native species' behaviour. We used a natural experiment of populations of native red squirrels Sciurus vulgaris that were colonized by the alien grey squirrel Sciurus carolinensis, comparing repeated measurements of red squirrel parasite infection and personality with those taken in sites where only the native species occurred. We explored two alternative hypotheses: (a) individual differences in personality traits (activity and/or sociability) of native red squirrel positively affect the probability of macroparasite spillover and thus the likelihood to acquire the alien's parasitic helminth Strongyloides robustus; (b) the combined effects of grey squirrel presence and parasite infection result in a reduction of costly personality traits (activity and/or exploration). Using data from 323 arena tests across three experimental (native species and IAS) and three control sites (only native species), we found negative correlations between native species' activity and infection with S. robustus in the sites invaded by the alien species. Activity was also negatively correlated with infection by its native helminth Trypanoxyuris sciuri but only when grey squirrels were present, while in the red-only sites there was no relationship of T. sciuri infection with any of the personality traits. Moreover, individuals that acquired S. robustus during the study reduced their activity after infection, while this was not the case for animals that remained uninfected. Our results show that parasite-mediated competition is costly, reducing activity in individuals of the native species, and altering the native host-native parasite relationships.

The price of being bold? Relationship between personality and endoparasitic infection in a tree squirrel

Individual variation in behaviour can contribute to the heterogeneous distribution of parasites among hosts for example by affecting the probability of encountering infective stages (larvae). Here, we investigated the relationship between host boldness/exploration tendency and gastro-intestinal helminth infection in invasive Eastern grey squirrels (Sciurus carolinensis). We used direct helminth counts, data rarely available in host-parasite studies that often used less reliable indirect estimates of infection. We predicted that bolder animals with a strong exploration tendency will have higher parasite load than shy, less explorative hosts. We examined two parameters of parasite infection: infection status and intensity of infection. Individual personality of 207 grey squirrels was assessed by capture-mark-recapture (CMR), calculating the trappability and trap diversity indices as estimates of boldness and exploration, respectively. Since both indices were strongly correlated, we used PCA to derive a single score (first component) which had a high value for bold, exploring animals. At the end of the study, 77 individuals were euthanized and gastro-intestinal helminths were identified and counted. Overall 73% of grey squirrels were infected by Strongyloides robustus with the intensity of infection varying from 1 to 86 worms (mean ± SE = 10.7 ± 2.1 helminths per host). We found that bolder, more explorative animals were more likely to be infected by S. robustus. However, host personality was not related to parasite intensity, which was instead positively associated with host body mass. Our results confirm that differences in personality-related host behaviour can influence the distribution of infections within host populations and stimulate further questions on whether such personality-parasite relationships may affect the invasion process.

An examination of endoparasites and fecal testosterone levels in flying squirrels (Glaucomys spp.) using high performance liquid chromatography-ultra-violet (HPLC-UV).

The immuno-competence hypothesis proposes that higher levels of testosterone increases the susceptibility to parasitism. Here we examined the testosterone levels in two species of flying squirrels (Glaucomys): one known to regularly host a nematode species (Strongyloides robustus) without ill effects (G. volans) and a closely related species that is considered negatively affected by the parasite. We quantified fecal testosterone levels in northern and southern flying squirrels (G. sabrinus, G. volans) with high-performance liquid chromatography-ultraviolet spectroscopy (HPLC-UV), and compared levels to endoparasites detected in individual squirrels. Qualitatively, we found highest levels of testosterone in male northern flying squirrels infected with Strongyloides robustus. This analytical approach represents an alternative and equally reliable method to using enzyme-linked immunosorbent assay (ELISA), for detecting and quantifying fecal testosterone levels.

Applying genetic methods to identify northern and southern flying squirrels and determine conservation needs

The two species of flying squirrels found in North America are the northern (Glaucomys sabrinus) and the southern (G. volans) flying squirrel. Both species have wide range distributions across North America, yet differences in their preferred habitat result in few areas of sympatry. Climate change however, has shifted the southern flying squirrel’s range limit further north, thus allowing the two species to encounter each other more frequently. Glaucomys volans is the more competitive of the two, and also carries a nematode parasite (Strongyloides robustus) proven to be deleterious towards its sister species. The northern incursion of the southern flying squirrel may therefore dislodge its sister species from its original distribution. With G. sabrinus considered endangered in different parts of North America, and G. volans listed as “concerned” in its northernmost limit, the two species require close monitoring. Morphological identification is often challenging for incomplete and juvenile specimens, therefore we developed a molecular identification protocol using mitochondrial and nuclear markers in tandem to help distinguish each species. These protocols were tested on Glaucomys individuals located elsewhere in Canada and the USA to ensure the validity of the designed primers. Moreover, employing our methods may detect F1 hybrids. To assess the current genetic portrait of G. sabrinus populations in Quebec before the potential invasion of its sister species, a preliminary population genetics study using seven microsatellite loci was also realized. The various conservation and management implications are discussed.

Climate‐driven range shifts are stochastic processes at a local level: two flying squirrel species in Maine

AbstractNumerous directional range shifts have been linked to climate change, but mechanistic explanations are frequently absent. In North America, southern flying squirrel (Glaucomys volans) range has expanded northward as a result of climate change, and northern flying squirrel (G. sabrinus) range has shifted north, likely in response to competition. We tracked both species abundances as well as climate, food availability, and forest structure over 31 yr, and found complete species turnover from G. sabrinus to G. volans in 18 yr. We tested multiple generalized least squares models of each species' abundance over time. The top model for G. sabrinus indicated that abundance was likely shaped by the arrival of G. volans, a competitor and known vector of a pathological nematode, food, and modest anthropogenic changes to forest structure; the top model for G. volans suggested that warm winters coinciding with high mast years likely facilitated the initial establishment of G. volans, whereas positive density dependence likely shaped subsequent population dynamics. The only significant change to local climate was an increase in minimum summer temperature, which would enhance the survival of Strongyloides robustus, the nematode carried by G. volans and detrimental to G. sabrinus. This complex suite of interactions suggests that at a local level, range shifts may be a largely stochastic process permitted by the temporal coincidence of multiple species‐ and site‐specific factors.

Biodiversity threats from outside to inside: effects of alien grey squirrel (Sciurus carolinensis) on helminth community of native red squirrel (Sciurus vulgaris)

Biological invasions are among the major causes of biodiversity loss worldwide, and parasites carried or acquired by invaders may represent an added threat to native species. We compared gastrointestinal helminth communities of native Eurasian red squirrels (Sciurus vulgaris) in the presence and absence of introduced Eastern grey squirrels (Sciurus carolinensis) to detect alterations induced by the alien species. In particular, we investigated whether spillover of a North American nematode Strongyloides robustus occurs and whether prevalence of a local parasite Trypanoxyuris sciuri in red squirrels is affected by grey squirrel presence. The probability of being infected by both parasites was significantly higher in areas co-inhabited by the alien species, where 61% of examined red squirrels (n = 49) were infected by S. robustus and 90% by T. sciuri. Conversely, in red-only areas, the two parasites infected only 5 and 70% of individuals (n = 60). Overall, our findings support the hypothesis that red squirrels acquire S. robustus via spillover from the alien congener and suggest that invaders' presence may also indirectly affect infection by local parasites through mechanisms diverse than spill-back and linked to the increased competitive pressure to which red squirrels are subjected. These results indicate that the impact of grey squirrel on red squirrels may have been underestimated and highlight the importance of investigating variation in macroparasite communities of native species threatened by alien competitors.

Faecal egg counts from field experiment reveal density dependence in helminth fecundity: Strongyloides robustus infecting grey squirrels (Sciurus carolinensis).

Investigation of endo-macroparasite infections in living animals relies mostly on indirect methods aimed to detect parasite eggs in hosts' faeces. However, faecal flotation does not provide quantitative information on parasite loads, whereas faecal egg count (FEC) techniques may not give reliable estimates of parasite intensity, since egg production may be affected by density-dependent effects on helminth fecundity. We addressed this issue using Eastern grey squirrels (Sciurus carolinensis) and their gastrointestinal nematode Strongyloides robustus to assess the performance of coprological techniques and to investigate factors affecting parasite fecundity. We compared results of gut examination, flotation and McMaster FECs in 65 culled grey squirrels. Sensitivity and specificity of flotation were 81.2% (Confidence Interval, CI 54.3-95.9%) and 85.7% (CI 72.7-94.1%), respectively, resulting in low positive predictive values when infection prevalence is low. Individual parasite fecundity (no. of eggs/adult female worm) was negatively affected by S. robustus intensity, leading to a non-linear relationship between parasite load and eggs/gram of faeces (EPG). As a consequence, whereas flotation may be a valid method to perform the first screening of infection status, FECs are not a reliable method to estimate S. robustus intensity, since diverse values of EPG may correspond to the same number of parasites. Neither the amount of analysed faeces nor the season had any effect on EPG, indicating that the observed reduction in helminth fecundity is likely caused exclusively by density-dependent processes such as competition among worms or host immune response.

Macroparasite Fauna of Alien Grey Squirrels (Sciurus carolinensis): Composition, Variability and Implications for Native Species

Introduced hosts populations may benefit of an "enemy release" through impoverishment of parasite communities made of both few imported species and few acquired local ones. Moreover, closely related competing native hosts can be affected by acquiring introduced taxa (spillover) and by increased transmission risk of native parasites (spillback). We determined the macroparasite fauna of invasive grey squirrels (Sciurus carolinensis) in Italy to detect any diversity loss, introduction of novel parasites or acquisition of local ones, and analysed variation in parasite burdens to identify factors that may increase transmission risk for native red squirrels (S. vulgaris). Based on 277 grey squirrels sampled from 7 populations characterised by different time scales in introduction events, we identified 7 gastro-intestinal helminths and 4 parasite arthropods. Parasite richness is lower than in grey squirrel's native range and independent from introduction time lags. The most common parasites are Nearctic nematodes Strongyloides robustus (prevalence: 56.6%) and Trichostrongylus calcaratus (6.5%), red squirrel flea Ceratophyllus sciurorum (26.0%) and Holarctic sucking louse Neohaematopinus sciuri (17.7%). All other parasites are European or cosmopolitan species with prevalence below 5%. S. robustus abundance is positively affected by host density and body mass, C. sciurorum abundance increases with host density and varies with seasons. Overall, we show that grey squirrels in Italy may benefit of an enemy release, and both spillback and spillover processes towards native red squirrels may occur.

Macroparasite community of the Eurasian red squirrel (Sciurus vulgaris): poor species richness and diversity

The Eurasian red squirrel (Sciurus vulgaris) is the only naturally occurring tree squirrel throughout its range. We aim at improving current knowledge on its macroparasite fauna, expecting that it will have a poor parasite diversity because in species that have no sympatric congeners parasite richness should be lower than in hosts sharing their range with several closely related species, where host-switching events and lateral transmission are promoted. We examined gastro-intestinal helminth and ectoparasite communities (excluding mites) of, respectively, 147 and 311 red squirrel roadkills collected in four biogeographic regions in Italy and France. As expected, the macroparasite fauna was poor: we found five species of nematodes and some unidentified cestodes, three fleas, two sucking lice and two hard ticks. The helminth community was dominated by a single species, the oxyurid Trypanoxyuris (Rodentoxyuris) sciuri (prevalence, 87%; mean abundance, 373 ± 65 worms/host). Its abundance varied among seasons and biogeographic regions and increased with body mass in male hosts while decreased in females. The most prevalent ectoparasites were the flea Ceratophyllus (Monopsyllus) sciurorum (28%), whose presence was affected by season, and the generalist tick Ixodes (Ixodes) ricinus that was found only in France (34%). All the other helminths and arthropod species were rare, with prevalence below 10%. However, the first record of Strongyloides robustus, a common nematode of North American Eastern grey squirrels (S. carolinensis), in two red squirrels living in areas where this alien species co-inhabits, deserves further attention, since low parasite richness could result in native red squirrels being particularly vulnerable to parasite spillover.

Prevalence of Strongyloides robustus in tree squirrels (Sciuridae) in South-Central Pennsylvania and potential impacts for the endangered northern flying squirrel, Glaucomys sabrinus.

Strongyloides robustus is a unique parasite that has conservation impacts for sciurid populations in North America. In some squirrel species, like the southern flying squirrel (Glaucomys volans), the eastern gray squirrel (Sciurus carolinensis) and the red squirrel (Tamiasciurus hudsonicus), pathology is relatively benign. However in the northern flying squirrel (Glaucomys sabrinus), S. robustus infestation can result in high mortality. The objective of this project was to survey the prevalence of S. robustus within the squirrel species currently found in south- central Pennsylvania so that the risk to the northern flying squirrel could be evaluated in light of the parasite mediated competition hypothesis. Fecal samples from eastern gray, red, and southern flying squirrels were obtained through nest boxes, road kills and hunting. A modified Sheather’s sugar floatation was prepared with a specific gravity of 1.27 to evaluate parasite prevalence. Ten of the 40 nest boxes examined had flying squirrel evidence in the form of feces deposited within the nest. Strongyloides robustus was present in 30% of the 10 samples. The prevalence of S. robustus was 77.3% in the 22 road-killed and hunter-killed eastern gray squirrels. The single hunter-killed red squirrel examined in this study demonstrated S. robustus infestation. This study evaluated infection in possible reservoirs that are understudied in Pennsylvania and supported the idea of parasite mediated competition.

THE POTENTIAL ROLE OF STRONGYLOIDES ROBUSTUS ON PARASITE-MEDIATED COMPETITION BETWEEN TWO SPECIES OF FLYING SQUIRRELS (GLAUCOMYS)

There is growing evidence that populations of the northern flying squirrel (Glaucomys sabrinus) are declining in the eastern United States, perhaps due to competition with the southern flying squirrel (Glaucomys volans). Potential causes include parasite-mediated or apparent competition from the shared intestinal nematode, Strongyloides robustus, which has been shown to detrimentally affect the northern flying squirrel but not the southern flying squirrel. To investigate this hypothesis, we conducted a preliminary study on the parasite community of both flying squirrel species from sites in Pennsylvania where the two species occur sympatrically and where G. sabrinus is now considered endangered at the state level. We compared these parasite communities with those from northern flying squirrels from northern New York where the southern flying squirrel is absent. We found eight species of gastrointestinal parasites (Pterygodermatites peromysci, Lemuricola sciuri, Syphacia thompsoni, Syphacia spp., Capillaria spp., Citellinema bifurcatum, Strogyloides robustus, and an unidentifiable cestode species) in both species of flying squirrels examined for our study. The parasite-mediated competition hypothesis was partially supported. For example, in Pennsylvania, S. robustus was overdispersed in southern flying squirrels, such that a small proportion of the hosts carried a large proportion of the worm population. In addition, we found S. robustus to be present in northern flying squirrels when the species were sympatric, but not where southern flying squirrels were absent in New York. However, there was no association between S. robustus and the body condition of flying squirrels. We detected a potential parasite community interaction, as S. robustus abundance was positively associated with P. peromysci.

A NEW RHABDITOID NEMATODE SPECIES IN ASIAN SCIURIDS, DISTINCT FROM STRONGYLOIDES ROBUSTUS IN NORTH AMERICAN SCIURIDS

Strongyloides callosciureus n. sp. (Nematoda: Rhabditoidea), from Asian sciurids, is described based on morphology, morphometry, and the small and large subunit (SSU/LSU) ribosomal RNA gene (rDNA) sequences. This new species was collected from Pallas's squirrels (Callosciurus erythraeus) in the central part of mainland Japan (Honshu), which were originally introduced from Taiwan some decades ago, and plantain squirrels (Callosciurus notatus) imported from Malaysia as personal pets. For comparison, Strongyloides robustus Chandler, 1942 was collected from American red squirrels (Tamiasciurus hudsonicus) and southern flying squirrels (Glaucomys volans) imported from the United States as personal pets. The parasitic females found in North American and Asian sciurids shared some key morphological features such as the ovary running spirally around the gut, and the shapes of the stoma in the apical view and the tail. However, morphometric features of parasitic females in North American and Asian sciurids differed significantly from each other; the former was larger than the latter, and the relative position of the vulva to the whole body length from the mouth was different. The SSU/LSU rDNA sequences supported the division of sciurid Strongyloides isolates by geographical distribution of the host and morphological features, leading us to propose the erection of new species.

Strongyloides robustus and the northern sympatric populations of northern (Glaucomys sabrinus) and southern (G. volans) flying squirrels.

Within North America, northern (Glaucomys sabrinus) and southern (Glaucomys volans) flying squirrels occupy distinct ranges with limited overlap. Sympatry in northern latitudes coincides with northern hardwood vegetation from Minnesota to New England. Strongyloides robustus is an intestinal parasite that infects both species but appears to be deleterious only to northern flying squirrels. As a result, S. robustus could be a critical determinant of flying squirrel population characteristics in at least some areas of sympatry. However, cold weather could potentially limit the distribution of S. robustus in northern climates. Therefore, we assessed fecal samples from both flying squirrel species to determine the presence of the nematode in Wisconsin. Strongyloides robustus was found in 12 flying squirrel scat samples and infected 52% of southern flying squirrels and 11% of northern flying squirrels. Prevalence of S. robustus infection for northern flying squirrels was substantially lower than previously reported from more southern regions. This is the northernmost documentation of S. robustus in flying squirrels and the first documentation of S. robustus parasitizing flying squirrels in Wisconsin.

Ecological Implications for Flying Squirrels (Glaucomys spp.) of Effects of Temperature on the In Vitro Development and Behavior of Strongyloides robustus

-The effects of temperature on in vitro egg-hatching success, juvenile survival, development and behavior of Strongyloides robustus were examined. As culture temperatures declined (25-10 C), both egg hatching and rate of larval development decreased. Hatching occurred as low as 10 C. Eggs showed low resistance to freezing temperatures and no juveniles survived freezing. When exposed to a thermal gradient, filariform juveniles moved toward the higher temperature. The mean temperature at which worms were found was 37.0 ? 3.3 C, which approximates host temperature within a nest. These results suggest that dissimilarities in the prevalences and intensities of infection with S. robustus in the northern and southern flying squirrels, Glaucomys sabrinus and G. volans, respectively, are at least partly due to environmental differences between habitats. Overlap in the squirrels' nest use may facilitate passage of this parasite from one host to the other, but the survival of eggs and infective juveniles may be severely limited by low temperatures. While transmission of S. robustus is limited during winter, summer transmission alone could account for the maintenance of the parasitic infections in these hosts.