Description and epidemiology of Theileria youngi n. sp. from a northern Californian dusky-footed woodrat (Neotoma Fuscipes) population

An epidemiologic study designed to identify the small mammal reservoir for the zoonotic WA1-type babesial parasite resulted in the discovery of a small, intraerythrocytic piroplasm in smeared blood from dusky-footed woodrats (Neotoma fuscipes) in northern California. The woodrat parasites were isolated and compared to other piroplasm parasites based on their morphology, antigenicity, and genetic characteristics. These studies indicated that the woodrat parasites were not the WA1-type babesial agent but were of the genus Theileria. We accordingly named it Theileria youngi. The prevalence in the woodrat population was high (61%). Infection was unrelated to gender or age of the woodrats. Potential vectors for this tick-transmitted parasite included 3 species of ticks recovered from the woodrats. Dermacentor occidentalis, Ixodes woodi, and Ixodes pacificus. Mostly larval or nymphal stages were recovered, suggesting transstadial transmission is possible. This is the first piroplasm fully characterized from a dusky-footed woodrat.     

Techniques for immobilizing and bleeding marmots and woodrats

Blood samples were obtained in the field by femoral vein puncture in bushy-tailed woodrats (Neotoma cinerea) and yellow-bellied marmots (Marmota flaviventris) that had been injected intramuscularly with ketamine hydrochloride. Dosages ranged from 50 mg/kg for marmots to 30 to 110 mg/kg for woodrats. Sedated animals were handled easily, and a volume of blood sufficient for hematological assays or electrophoresis could be collected.     

Epizootiology of arenaviral infections in the white-throated woodrat (Muridae: Sigmodontinae) and other woodrats in Arizona.

The purpose of this study was to extend and refine our knowledge of the geographical distribution and natural host relationships of the arenaviruses associated with woodrats indigenous to Arizona. Antibody to a Tacaribe serocomplex virus was found in 112 of 1,250 white-throated woodrats, five of 208 Mexican woodrats, one of 114 Stephen's woodrats, and none of 862 other rodents captured at 51 sites in 10 counties in Arizona. Of the 112 antibody-positive white-throated woodrats, 109 (97.3%) were captured within extensive, dense patches of prickly pear cactus and cane cholla in three counties in mid-central Arizona. Analysis of the serological and zoographical data suggested that white-throated woodrats usually become infected early in life and that the distribution of antibody-positive white-throated woodrats in Arizona is not linked to a specific biome.     

Plant secondary metabolites alter the feeding patterns of a mammalian herbivore (<Emphasis Type="Italic">Neotoma lepida</Emphasis>)

Mammalian herbivores are predicted to regulate concentrations of ingested plant secondary metabolites (PSMs) in the blood by modifying the size and frequency of feeding bouts. It is theorized that meal size is limited by a maximum tolerable concentration of PSMs in the blood, such that meal size is predicted to decrease as PSM concentration increases. We investigated the relationship between PSM concentration in the diet and feeding patterns in the herbivorous desert woodrat (Neotoma lepida) fed diets containing phenolic resin extracted from creosote bush (Larrea tridentata). Total daily intake, meal size and feeding frequency were quantified by observing the foraging behavior of woodrats on diets containing increasing concentrations of creosote resin. Desert woodrats reduced meal size as resin concentration in the diet increased, resulting in an overall reduction in daily intake and regulation of resin intake. Moreover, desert woodrats were able to detect resin concentrations in the diet and regulate the intake of resin very rapidly. We suggest that the immediate and sustained ability to detect and regulate the intake of resin concentrations during each foraging bout provides a behavioral mechanism to regulate blood concentrations of resin and allows desert woodrats to make “wise” foraging decisions.     

Expression of biotransformation genes in woodrat (Neotoma) herbivores on novel and ancestral diets: identification of candidate genes responsible for dietary shifts

The ability of herbivores to switch diets is thought to be governed by biotransformation enzymes. To identify potential biotransformation enzymes, we conducted a large-scale study on the expression of biotransformation enzymes in herbivorous woodrats ( Neotoma lepida ). We compared gene expression in a woodrat population from the Great Basin that feeds on the ancestral diet of juniper to one from the Mojave Desert that putatively switched from feeding on juniper to feeding on creosote. Juniper and creosote have notable differences in secondary chemistry, and thus, should require different biotransformation enzymes for detoxification. Individuals from each population were fed juniper and creosote diets separately. After the feeding trials, hepatic mRNA was extracted and hybridized to laboratory rat microarrays. Hybridization of woodrat samples to biotransformation probes on the array was 87%, resulting in a total of 224 biotransformation genes that met quality control standards. Overall, we found large differences in expression of biotransformation genes when woodrats were fed juniper vs. creosote. Mojave woodrats had greater expression of 10× as many biotransformation genes as did Great Basin woodrats on a creosote diet. We identified 24 candidate genes that may be critical in the biotransformation of creosote toxins. Superoxide dismutase, a free radical scavenger, was also expressed to a greater extent by the Mojave woodrats and may be important in controlling oxidative damage during biotransformation. The results are consistent with the hypothesis that biotransformation enzymes limit diet switching and that woodrats in the Mojave have evolved a unique strategy for the biotransformation of creosote toxins.     

Caching behaviour by eastern woodrats,Neotoma floridana,in relation to food perishability

Two experiments were conducted on eastern woodrats to determine whether the preference for food items gathered for their caches and the sequence of use of items in the cache varied with perishability. In the first experiment, woodrats were given either 2 kg each of two non-perishable items, laboratory chow and bur oak acorns (which they used more than chow), or 2 kg each of preferred, but perishable, grapes and the non-perishable chow. As hypothesized, the woodrats both ate and stored acorns when they were paired with chow (there would be no advantage to either eating or storing the less-used chow). In contrast, when given grapes and chow, the woodrats continued to eat grapes but stored primarily chow. That is, the utility of the items appeared to change depending on whether they were to be eaten or stored. In the second experiment, woodrats were given excess amounts of chow or grapes that were 2, 4, 6 and 8 days old. They ingested chow in the proportions available, as the chow did not spoil with age. However, the consumed grapes in proportions significantly different from those available. It was anticipated that they would eat the oldes grapes first and store the freshest grapes, but neither this pattern nor any other was obvious in ralation to the age of the grapes. The results suggest that woodrats discriminate between food items based on their perishability, and vary their decisions about what to ingest and store accordingly.     

Eimeria ladronensis n. sp. and E. albigulae (Apicomplexa: Eimeriidae) from the woodrat, Neotoma albigula (Rodentia: Cricetidae)

Of 50 white-throated woodrats (Neotoma albigula) collected from Socorro Co., New Mexico, 21 (42%) had eimerian oocysts in their feces when examined. Of the 21 Neotoma found positive for Eimeria, 19 (90%) harbored a single eimerian species at time of examination. Eimeria albigulae Levine, Ivens & Kruidenier, 1957, was found in 18 (86%), and E. ladronensis n. sp. was found in five (24%) infected woodrats. Sporulated oocysts of E. ladronensis are ellipsoidal, 19-25 X 13-15 (21.4 +/- 1.3 X 14.1 +/- 1.1) micron, have a smooth wall and one or two polar granules, but lack a micropyle and an oocyst residuum. Sporocysts are tapered at one end, 7-10 X 6-7 (8.5 +/- 0.7 X 6.5 +/- 0.3) micron, and have a Stieda body and sporocyst residuum, but no substieda body. Prepatent periods for E. albigulae and E. ladronensis n. sp. are 5-6 and 8-9 days, respectively; patent periods are 7-18 and approximately 11 days, respectively.     

Ambient temperature influences diet selection and physiology of an herbivorous mammal, Neotoma albigula

The whitethroat woodrat (Neotoma albigula) eats juniper (Juniperus monosperma), but the amount of juniper in its diet varies seasonally. We tested whether changes in juniper consumption are due to changes in ambient temperature and what the physiological consequences of consuming plant secondary compounds (PSCs) at different ambient temperatures might be. Woodrats were acclimated to either 20 degrees C or 28 degrees C. Later, they were given two diets to choose from (50% juniper and a nontoxic control) for 7 d. Food intake, resting metabolic rate (RMR), and body temperature (T(b)) were measured over the last 2 d. Woodrats at 28 degrees C ate significantly less juniper, both proportionally and absolutely, than woodrats at 20 degrees C. RMRs were higher for woodrats consuming juniper regardless of ambient temperature, and T(b) was higher for woodrats consuming juniper at 28 degrees C than for woodrats eating control diet at 28 degrees C. Thus, juniper consumption by N. albigula is influenced by ambient temperature. We conclude that juniper may influence thermoregulation in N. albigula in ways that are helpful at low temperatures but harmful at warmer temperatures in that juniper PSCs may be more toxic at warmer temperatures. The results suggest that increases in ambient temperature associated with climate change could significantly influence foraging behavior of mammalian herbivores.     

Influence of landscape heterogeneity on the functional connectivity of Allegheny woodrats (<Emphasis Type="Italic">Neotoma magister</Emphasis>) in Virginia

Allegheny woodrats (Neotoma magister) exist as groups of metapopulations due to their dependence on naturally disjunct rocky outcrops in the eastern United States. Severe demographic declines of Allegheny woodrats have occurred in many parts of the range due to a myriad of interacting processes, therefore identifying factors that help maintain the integrity of metapopulations is needed to guide conservation efforts. One factor considered critical to metapopulation persistence is maintaining functional connectivity. Therefore, our objective was to identify landscape factors that influence gene flow in Allegheny woodrats. We sampled 159 individuals from throughout the distribution in Virginia, genotyped them at 22 microsatellite loci, and modeled effects of land cover, roads, large rivers, and elevation on gene flow. The model that parameterized areas?≤?750 m in elevation as a barrier to gene flow performed best among the tested models, which indicates that functional connectivity is highest in high elevation habitats. We found little evidence for anthropogenic factors influencing gene flow, but continued study across extant metapopulations is needed in more peripheral areas to evaluate if anthropogenic barriers impact functional connectivity. Overall, our study emphasizes the importance of considering elevation for maintaining functional connectivity of Allegheny woodrats in the face of ongoing demographic declines in the eastern United States.     

Transmission cycles of Borrelia burgdorferi and B. bissettii in relation to habitat type in northwestern California

This study was undertaken to determine which rodent species serve as primary reservoirs for the Lyme disease spirochete Borrelia burgdorferi in commonly occurring woodland types in inland areas of northwestern California, and to examine whether chaparral or grassland serve as source habitats for dispersal of B. burgdorferi‐ or B. bissettii‐infected rodents into adjacent woodlands. The western gray squirrel (Sciurus griseus) was commonly infected with B. burgdorferi in oak woodlands, whereas examination of 30 dusky‐footed woodrats (Neotoma fuscipes) and 280 Peromyscus spp. mice from 13 widely‐spaced Mendocino County woodlands during 2002 and 2003 yielded only one infected woodrat and one infected deer mouse (P. maniculatus). These data suggest that western gray squirrels account for the majority of production by rodents of fed Ixodes pacificus larvae infected with B. burgdorferi in the woodlands sampled. Infections with B. burgdorferi also were rare in woodrats (0/47, 0/3) and mice (3/66, 1/6) captured in chaparral and grassland, respectively, and therefore these habitats are unlikely sources for dispersal of this spirochete into adjacent woodlands. On the other hand, B. bissettii was commonly detected in both woodrats (22/47) and mice (15/66) in chaparral. We conclude that the data from this and previous studies in northwestern California are suggestive of a pattern where inland oak‐woodland habitats harbor a B. burgdorferi transmission cycle driven primarily by I. pacificus and western gray squirrels, whereas chaparral habitats contain a B. bissettii transmission cycle perpetuated largely by I. spinipalpis, woodrats, and Peromyscus mice. The dominant role of western gray squirrels as reservoirs of B. burgdorferi in certain woodlands offers intriguing opportunities for preventing Lyme disease by targeting these animals by means of either host‐targeted acaricides or oral vaccination against B. burgdorferi.     

Evidence for local specialization in a generalist mammalian herbivore, Neotoma fuscipes

Herbivores with very plastic dietary requirements, or so-called generalist species, can include individuals that develop specialized feeding habits through their experience with local chemically-defended plants. Local specialization has important implications for understanding a variety of ecological and evolutionary dynamics. However, the extent to which individuals within a generalist species specialize on local plants and the consequences of such specialization remain poorly understood, especially in non-insect herbivores. To better understand this phenomenon, we determined the diet and food preferences of a generalist mammalian herbivore, the dusky-footed woodrat ( Neotoma fuscipes ), in two adjacent but distinct plant communities. Based on a combination of cafeteria trials and stable isotope analyses (δ¹³C and δ¹⁵N), our results indicate that woodrats display preferences for local plants and tend to avoid novel chemically-defended plants. Moreover, both methodologies support the conclusion that individual woodrats are dietary specialists restricting their diets to only a few (2-3) of the available plant species. In juniper woodland, woodrats prefer western juniper ( Juniperus occidentalis ), while less than one km away in mixed-coniferous forest, woodrats prefer incense cedar ( Calocedrus decurrens ). Both plants contain high levels of plant secondary compounds that require detoxification mechanisms within consumers. Therefore, preferences are likely indicative of underlying physiological adaptations that could promote further behavioral, physiological and ultimately genetic differences between woodrats in different habitats. This study provides additional evidence for local specialization and urges caution when using the term generalist to characterize feeding behaviors at the individual level.     

Ambient temperature influences tolerance to plant secondary compounds in a mammalian herbivore

Growing evidence suggests that plant secondary compounds (PSCs) ingested by mammals become more toxic at elevated ambient temperatures, a phenomenon known as temperature-dependent toxicity. We investigated temperature-dependent toxicity in the desert woodrat (Neotoma lepida), a herbivorous rodent that naturally encounters PSCs in creosote bush (Larrea tridentata), which is a major component of its diet. First, we determined the maximum dose of creosote resin ingested by woodrats at warm (28–29°C) or cool (21–22°C) temperatures. Second, we controlled the daily dose of creosote resin ingested at warm, cool and room (25°C) temperatures, and measured persistence in feeding trials. At the warm temperature, woodrats ingested significantly less creosote resin; their maximum dose was two-thirds that of animals at the cool temperature. Moreover, woodrats at warm and room temperatures could not persist on the same dose of creosote resin as woodrats at the cool temperature. Our findings demonstrate that warmer temperatures reduce PSC intake and tolerance in herbivorous rodents, highlighting the potentially adverse consequences of temperature-dependent toxicity. These results will advance the field of herbivore ecology and may hone predictions of mammalian responses to climate change.     

Elimination of plant toxins by herbivorous woodrats: revisiting an explanation for dietary specialization in mammalian herbivores

Constraints on rates of detoxification and elimination of plant toxins are thought to be responsible for limiting dietary specialization in mammalian herbivores. This hypothesis, known as the detoxification limitations hypothesis, suggests that most mammalian herbivores are generalists to avoid overdosing on toxins from a single plant species. The hypothesis also predicts that the few mammalian specialists that exist should have adaptations for rapid detoxification and elimination of plant secondary compounds. We took a pharmacological approach to test whether specialists eliminate toxins from the bloodstream faster than generalists. We compared elimination rate and total exposure of alpha-pinene in closely related dietary specialist and generalist woodrats, Neotoma stephensi and N. albigula, respectively. Animals were orally gavaged with alpha-pinene, a plant secondary compound present in the natural diets of both woodrat species. We collected venous blood at 3, 6, 10, 15, and 20 min post-ingestion of alpha-pinene. Blood was analyzed for alpha-pinene concentration using gas chromatography. We found that specialist and generalist woodrats did not differ in elimination rates of alpha-pinene. However, specialists had lower exposure levels of alpha-pinene than generalists due to lower initial delivery of alpha-pinene to the general circulation. The levels of alpha-pinene detected in the bloodstream of specialists were 4.7-5.3x lower over all time intervals than generalists. Thus, specialists encounter a functionally lower dose of toxin than generalists. We suggest that the lower exposure level of specialist woodrats may be due to mechanisms in the gut that decrease toxin absorption. Regardless of mechanism, lower exposure to plant toxins may allow specialists to forage on diets with high toxin concentrations thereby facilitating dietary specialization.     

Implications of Woodrats and Other Scavengers for Avian Telemetry Studies

Abstract: During 2 years of radiotelemetry research on chukars (Alectoris chukar) in western Utah, USA, we found 28% of retrieved radios (n = 78) in rubbish nests of woodrats (Neotoma spp.). Such movement and disturbance of carcasses and radios by woodrats and other species has implications for radiotelemetry studies. We evaluated spatio-temporal movement of 51 radiocollars attached to chukar carcasses in western Utah. Most (80%) carcasses were scavenged within one week and by the end of 3 weeks 25 (50%) had been retrieved from woodrat middens. Scavenging activity can both obscure important clues needed to identify causes of mortality and bias telemetry studies by delaying onset of mortality signals.     

Eimeria ladronensis n. sp. and E. albigulae (Apicomplexa: Eimeriidae) from the Woodrat,Neotoma albigula (Rodentia: Cricetidae)1

Of 50 white-throated woodrats (Neotoma albigula) collected from Socorro Co., New Mexico, 21 (42%) had eimerian oocysts in their feces when examined. Of the 21 Neotoma found positive for Eimeria, 19 (90%) harbored a single eimerian species at time of examination. Eimeria albigulae Levine, Ivens & Kruidenier, 1957, was found in 18 (86%), and E. ladronensis n. sp. was found in five (24%) infected woodrats. Sporulated oocysts of E. ladronensis are ellipsoidal, 19–25 × 13–15 (21.4 ± 1.3 × 14.1 ± 1.1) μm, have a smooth wall and one or two polar granules, but lack a micropyle and an oocyst residuum. Sporocysts are tapered at one end, 7–10 × 6–7 (8.5 ± 0.7 × 6.5 ± 0.3) μm, and have a Stieda body and sporocyst residuum, but no substieda body. Prepatent periods for E. albigulae and E. ladronensis n. sp. are 5–6 and 8–9 days, respectively; patent periods are 7–18 and approximately 11 days, respectively.     

Experience matters: prior exposure to plant toxins enhances diversity of gut microbes in herbivores

For decades, ecologists have hypothesised that exposure to plant secondary compounds (PSCs) modifies herbivore‐associated microbial community composition. This notion has not been critically evaluated in wild mammalian herbivores on evolutionary timescales. We investigated responses of the microbial communities of two woodrat species (Neotoma bryanti and N. lepida). For each species, we compared experienced populations that independently converged to feed on the same toxic plant (creosote bush, Larrea tridentata) to naïve populations with no exposure to creosote toxins. The addition of dietary PSCs significantly altered gut microbial community structure, and the response was dependent on previous experience. Microbial diversity and relative abundances of several dominant phyla increased in experienced woodrats in response to PSCs; however, opposite effects were observed in naïve woodrats. These differential responses were convergent in experienced populations of both species. We hypothesise that adaptation of the foregut microbiota to creosote PSCs in experienced woodrats drives this differential response.     

The roles of birds, lizards, and rodents as hosts for the western black-legged tick Ixodes pacificus.

We compared the infestation by ixodid ticks of lizards, rodents, and birds collected simultaneously within areas representing common habitat types in Mendocino County, CA. Lizards were infested only by Ixodes pacificus Cooley and Kohls, birds by I. pacificus and Haemaphysalis leporispalustris (Packard), and rodents by I. pacificus, I. spinipalpis Hadwen and Nuttall, I. woodi Bishopp, Dermacentor occidentalis Marx, and D. variabilis (Say). Infestation by I. pacificus larvae and nymphs of lizards (Sceloporus occidentalis Baird and Girard; Elgaria spp.) and western gray squirrels (Sciurus griseus Ord) (means of 9-35 larvae and 5-6 nymphs per animal) was several times greater than for Neotoma fuscipes Baird woodrats, Peromyscus spp. mice, and birds (means of 0.9-3.5 larvae and 0-0.3 nymphs). Overall, Borrelia-refractory lizards accounted for 84% of I. pacificus larvae and 91% of nymphs collected from animals in dense woodlands. Bird species frequently utilizing tick-questing substrates such as leaf litter (guild I birds) were more heavily infested by I. pacificus subadults (5.2 larvae and 1.0 nymphs per bird) than guild IV birds with minimal perceived contact with tick-questing substrates (0.08 larvae and 0.06 nymphs per bird). Notably, guild I birds carried similar larval loads and at least 20-fold higher nymphal loads relative to woodrats and mice. Only guild IV birds carried as few I. pacificus nymphs as did these rodents. The ratios of larvae to nymphs suggest that, relative to birds, lizards, and squirrels (infested by 1.3-6.0 larvae per nymph), nocturnally active ground-dwelling rodents such as woodrats and mice are underutilized by the nymphal stage (69 to >100 larvae per nymph). The western gray squirrel and guild I-II birds (e.g., the dark-eyed junco, Junco hyemalis [L.]) were the only potential reservoirs of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt, and Brenner (the causative agent of Lyme disease in North America) that were frequently infested with both I. pacificus larvae and nymphs and commonly utilized dense woodland habitats.     

Survey of raccoons on Key Largo, Florida, USA, for Baylisascaris procyonis

Numbers of the endangered Key Largo woodrat (KLWR; Neotoma floridana smalli) have been declining for at least 25 yr. The raccoon (Procyon lotor) roundworm, Baylisascaris procyonis, has been found to have an adverse effect on the survival of Alleghany woodrats (N. magister). High densities of raccoons can exacerbate this problem by increasing the amount of feces containing viable eggs of B. procyonis available to woodrats. In 2002, 64 fecal samples were collected and examined for eggs of B. procyonis from >32 raccoons within the KLWR's known range on Key Largo, Florida, USA. All samples were negative for eggs of B. procyonis. Raccoon density in this area was approximately 0.62 raccoons/ha. Despite this high density of raccoons, B. procyonis does not appear to be a threat to the KLWR population.     

Field Use of Isoflurane for Safe Squirrel and Woodrat Anesthesia

Abstract: We evaluated a chamber and nose cone method of isoflurane delivery for anesthetizing eastern gray squirrels (Sciurus carolinensis; summer n = 43, winter n = 48) and Allegheny woodrats (Neotoma magister; summer n = 24, winter n = 13) for use when pain or stress was possible from sampling procedures. Mean induction time for squirrels (from beginning of isoflurane administration to safe removal from trap), was 4.63 ± 0.58 minutes. Squirrels awoke more quickly in summer (1.40 ± 0.15 min) than in winter (3.62 ± 0.24 min) after removal of the nose cone. We manually restrained woodrats and administered the nose cone for 0.5 minutes to each animal. Woodrats awoke after 4.76 ± 0.58 minutes following the final dose of isoflurane for both seasons. These methods are useful for working with small mammals in the field and provide an appropriate anesthetic when there may be more than slight pain or distress.     

Ingestion of plant secondary compounds causes diuresis in desert herbivores

Plant secondary compounds are recognized deterrents and toxins to a variety of herbivores. The effect of secondary compounds on water balance of herbivores is virtually unexplored, yet secondary compounds could potentially cause a decrease in an animal's ability to maintain water balance. We investigated the effects of secondary compounds, alpha-pinene and creosote resin, on water balance in three species of herbivorous woodrats (Neotoma stephensi, N. albigula, N. lepida). In separate experiments, we measured the effect of these secondary compounds on voluntary water consumption, urine volume and urine osmolarity. In both experiments, water intake and urine volume increased and urine osmolarity decreased compared to controls. Water balance of specialist or experienced woodrats was less affected than generalists and woodrats with less prior experience with particular secondary compounds. Our results suggest that secondary compounds have diuretic-like effects on herbivores. Woodrats live in arid habitats with limited access to freestanding water; thus an increase in water requirements may have profound consequences on foraging behavior and fitness.