Qiviut cortisol reflects hypothalamic-pituitary-adrenal axis activity in muskoxen (Ovibos moschatus).

Muskoxen (Ovibos moschatus) are increasingly exposed to a broad diversity of stressors in their rapidly changing Arctic environment. There is an urgent need to develop validated tools to monitor the impact of these stressors on the hypothalamic-pituitary-adrenal (HPA) axis activity of muskoxen to help inform conservation actions. Here, we evaluated whether muskox qiviut (dense wooly undercoat) cortisol accurately reflects changes in HPA axis activity. Two repeated pharmacological challenges, involving weekly administrations of saline (control group) or adrenocorticotropic hormone (ACTH) during five consecutive weeks, were done on captive muskoxen, in winter (no hair growth) and summer (maximum hair growth). Pre-challenge qiviut cortisol levels were significantly higher in the shoulder than in the neck, but neither differed from rump concentrations. Qiviut cortisol levels significantly increased (p < 0.001) in response to the administration of ACTH during the hair growth phase, but not in the absence of growth (p = 0.84). Cortisol levels in the qiviut segment grown during the summer challenge increased significantly over a six-month period in the ACTH-injected muskoxen with a similar trend occurring in the control animals. Finally, cortisol levels in shed qiviut were significantly higher and not correlated to those of fully grown qiviut shaved three months earlier. Our results show that cortisol is deposited in qiviut during its growth and that qiviut cortisol can thus be used as an integrated measure of HPA axis activity over the period of the hair's growth. Differences in qiviut cortisol across body regions, significant differences in qiviut segments over time, and differences between shed qiviut versus unshed qiviut, highlight the importance of consistent design and methodology for sample collection and analyses in order to account for sources of variation when using qiviut cortisol as a biomarker of HPA axis activity in muskoxen.

Arctic systems in the Quaternary: ecological collision, faunal mosaics and the consequences of a wobbling climate.

Climate oscillations and episodic processes interact with evolution, ecology and biogeography to determine the structure and complex mosaic that is the biosphere. Parasites and parasite-host assemblages are key components in a general explanatory paradigm for global biodiversity. We explore faunal assembly in the context of Quaternary time frames of the past 2.6 million years, a period dominated by episodic shifts in climate. Climate drivers cross a continuum from geological to contemporary timescales and serve to determine the structure and distribution of complex biotas. Cycles within cycles are apparent, with drivers that are layered, multifactorial and complex. These cycles influence the dynamics and duration of shifts in environmental structure on varying temporal and spatial scales. An understanding of the dynamics of high-latitude systems, the history of the Beringian nexus (the intermittent land connection linking Eurasia and North America) and downstream patterns of diversity depend on teasing apart the complexity of biotic assembly and persistence. Although climate oscillations have dominated the Quaternary, contemporary dynamics are driven by tipping points and shifting balances emerging from anthropogenic forces that are disrupting ecological structure. Climate change driven by anthropogenic forcing has supplanted a history of episodic variation and is eliminating ecological barriers and constraints on development and distribution for pathogen transmission. A framework to explore interactions of episodic processes on faunal structure and assembly is the Stockholm Paradigm, which appropriately shifts the focus from cospeciation to complexity and contingency in explanations of diversity.

Variation of hair cortisol in two herds of migratory caribou (Rangifer tarandus): implications for health monitoring.

Migratory caribou (Rangifer tarandus sspp.) is an ecotype of conservation concern that is experiencing increased cumulative stressors associated with rapid climate change and development in Arctic Canada. Increasingly, hair cortisol concentrations (HCCs) are being used to monitor seasonal hypothalamic-pituitary-adrenal axis activity of ungulate populations; yet, the effect of key covariates for caribou (sex, season, sampling source, body location) are largely unknown. The objectives of this research were 4-fold: first, we assessed the impact of body location (neck, rump) sampling sites on HCC; second, we assessed key covariates (sex, sampling method, season) impacting HCCs of caribou; third, we investigated inter-population (Dolphin and Union (DU), Bluenose-East (BNE)) and inter-annual differences in HCC and fourth, we examined the association between HCCs and indices of biting insect activity on the summer range (oestrid index, mosquito index). We examined hair from 407 DU and BNE caribou sampled by harvesters or during capture-collaring operations from 2012 to 2020. Linear mixed-effect models were used to assess the effect of body location on HCC and generalized least squares regression (GLS) models were used to examine the impacts of key covariates, year and herd and indices of biting insect harassment. HCC varied significantly by body location, year, herd and source of samples (harvester vs capture). HCC was higher in samples taken from the neck and in the DU herd compared with the BNE, decreased linearly over time and was higher in captured versus hunted animals (P < 0.05). There was no difference in HCC between sexes, and indices of biting insect harassment in the previous year were not significantly associated with HCC. This study identifies essential covariates impacting the HCC of caribou that must be accounted for in sampling, monitoring and data interpretation.

Adaptations, life-history traits and ecological mechanisms of parasites to survive extremes and environmental unpredictability in the face of climate change.

Climate change is increasing weather unpredictability, causing more intense, frequent and longer extreme events including droughts, precipitation, and both heat and cold waves. The performance of parasites, and host-parasite interactions, under these unpredictable conditions, are directly influenced by the ability of parasites to cope with extremes and their capacity to adapt to the new conditions. Here, we review some of the structural, behavioural, life history and ecological characteristics of parasitic nematodes that allow them to persist and adapt to extreme and changing environmental conditions. We focus primarily, but not exclusively, on parasitic nematodes in the Arctic, where temperature extremes are pronounced, climate change is happening most rapidly, and changes in host-parasite interactions are already documented. We discuss how life-history traits, phenotypic plasticity, local adaptation and evolutionary history can influence the short and long term response of parasites to new conditions. A detailed understanding of the complex ecological processes involved in the survival of parasites in extreme and changing conditions is a fundamental step to anticipate the impact of climate change in parasite dynamics.

Renal trace elements in barren-ground caribou subpopulations: Temporal trends and differing effects of sex, age and season.

Caribou (Rangifer tarandus) are a culturally significant food resource for communities in northern Canada and Greenland. Many barren-ground caribou subpopulations are currently in decline, some dramatically; understanding the influence of stressors, such as toxic trace metals, is important. These contaminants enter Arctic terrestrial environments via atmospheric transport from industrialized areas and from local sources, accumulating there in the environment. Understanding how trace element concentrations interact and are influenced by caribou sex, age and season of collection is essential to evaluating trends in these elements over time and differences among subpopulations. We used path analysis to model the direct and indirect relationships between these variables in the Porcupine subpopulation and in barren-ground caribou from the Canadian Arctic and Greenland. Renal cadmium (Cd), copper (Cu) and mercury (Hg) varied significantly among subpopulations. Hg was positively correlated with Cd, Cu and selenium (Se) in female Porcupine caribou whereas Cd and Cu were negatively correlated in male Porcupine caribou. Age, season and sex influenced all three element concentrations and should be considered when comparing elements among caribou subpopulations or years. Renal Cd decreased slightly from the Canadian Western Arctic to Greenland and increased slightly over time, possibly reflecting patterns of atmospheric deposition. Renal Hg did not change significantly over time, and differences among subpopulations did not follow specific geographical patterns. Renal Cu declined over time, the changes being markedly different among subpopulations, sexes and seasons. This temporal decline is likely due to changes in diet, which could be driven by various environmental factors. Declining Cu concentrations in caribou is of concern as low levels could negatively affect reproductive success and therefore caribou at a population level. Continuing to monitor element concentrations in caribou is essential to better comprehend potential threats facing the species, and to promote food security in communities harvesting this important resource.

Multi-pathogen serological survey of migratory caribou herds: A snapshot in time.

Pathogens can impact host survival, fecundity, and population dynamics even when no obvious disease is observed. Few baseline data on pathogen prevalence and diversity of caribou are available, which hampers our ability to track changes over time and evaluate impacts on caribou health. Archived blood samples collected from ten migratory caribou herds in Canada and two in Greenland were used to test for exposure to pathogens that have the potential to effect population productivity, are zoonotic or are emerging. Relationships between seroprevalence and individual, population, and other health parameters were also examined. For adult caribou, the highest overall seroprevalence was for alphaherpesvirus (49%, n = 722), pestivirus (49%, n = 572) and Neospora caninum (27%, n = 452). Lower seroprevalence was found for parainfluenza virus type 3 (9%, n = 708), Brucella suis (2%, n = 758), and Toxoplasma gondii (2%, n = 706). No animal tested positive for antibodies against West Nile virus (n = 418) or bovine respiratory syncytial virus (n = 417). This extensive multi-pathogen survey of migratory caribou herds provides evidence that caribou are exposed to pathogens that may have impacts on herd health and revealed potential interactions between pathogens as well as geographical differences in pathogen exposure that could be linked to the bio-geographical history of caribou. Caribou are a keystone species and the socio-economic cornerstone of many indigenous cultures across the North. The results from this study highlight the urgent need for a better understanding of pathogen diversity and the impact of pathogens on caribou health.

Pathogens of domestic and free-ranging ungulates: global climate change in temperate to boreal latitudes across North America.

In North America broad-based research networks explore the interaction of vertebrates, their characteristic arrays of pathogens and emergent disease. A diversity of programmes address the impact of environmental change on animal health, zoonoses, and human health, but as yet no comprehensive framework or strategy has emerged to develop and implement policy and planning. In a regime of climate change and ecological perturbation, the need to document and understand the health, agricultural, societal and economic impact of pathogens and emerging infectious disease is urgent. An integrated and proactive planning process linking national and international resources can lead to informed predictions aboutthe impact of environmental change and can identify pathways for potential management and mitigation. An effective and comprehensive programme will have components for establishing priorities, developing primary data for faunal structure and biodiversity, a capacity for monitoring and surveillance (including scanning and targeted activities), and linkage to historical and contemporary baselines (against which to assess change) established through archival biological collections. Field and laboratory studies are also necessary to determine developmental thresholds, tolerances and tipping points for many pathogens to establish a context for recognising current constraints and future perturbation, and to explore factors that promote emergence for a variety of pathogens, vectors and pest species. Predictive modelling and risk assessment utilising a range of scenarios for climate change is a final step in this multidisciplinary process.

Protostrongylid parasites and pneumonia in captive and wild thinhorn sheep (Ovis dalli).

We describe health significance of protostrongylid parasites (Parelaphostrongylus odocoilei and Protostrongylus stilesi) and other respiratory pathogens in more than 50 naturally infected Dall's sheep (Ovis dalli dalli) from the Mackenzie Mountains, Northwest Territories (1998-2002) as well as in three Stone's sheep (O. d. stonei) experimentally infected with P. odocoilei (2000-2002). Histological lesions in the brain and distribution of P. odocoilei in the muscles of experimentally and naturally infected sheep were consistent with a previously hypothesized "central nervous system to muscle" pattern of migration for P. odocoilei. Dimensions of granulomas associated with eggs of P. odocoilei and density of protostrongylid eggs and larvae in the cranial lung correlated with intensity of larvae in feces, and all varied with season of collection. Prevalence of P. stilesi based on the presence of larvae in feces underestimated true prevalence (based on examination of lungs) in wild Dall's sheep collected in summer and fall. Similarly, counts of both types of protostrongylid larvae in feces were unreliable indicators of parasitic infection in wild Dall's sheep with concomitant bacterial pneumonia associated with Arcanobacterium pyogenes, Pasteurella sp., and Mannheimia sp. Diffuse, interstitial pneumonia due to P. odocoilei led to fatal pulmonary hemorrhage and edema after exertion in one experimentally infected Stone's sheep and one naturally infected Dall's sheep. Bacterial and verminous pneumonia associated with pathogens endemic in wild Dall's sheep in the Mackenzie Mountains caused sporadic mortalities. There was no evidence of respiratory viruses or bacterial strains associated with domestic ruminants, from which this population of wild sheep has been historically isolated.

Experimental Life-Cycle of Varestrongylus eleguneniensis (Nematoda: Protostrongylidae) in a Captive Reindeer (Rangifer tarandus tarandus) and a Muskox (Ovibos moschatus moschatus).

The life-cycle of a recently described protostrongylid lungworm, Varestrongylus eleguneniensis, which infects caribou, muskoxen, and moose from Arctic and boreal regions of North America, was completed experimentally for the first time. A native North American slug species, Deroceras laeve, was infected with the first-stage larvae (L1) isolated from the feces of wild muskoxen to generate third-stage larvae (L3). These were administered to a captive reindeer calf (250 L3) and an adult captive muskox (380 L3). The prepatent periods for the reindeer and muskox were 56 and 72 days, respectively. Patency lasted for only 19 days in the reindeer, and fecal larval counts were very low (0.09-1.53 larvae per gram of feces). Patency in the muskox was at least 210 days, and likely over 653 days, and the fecal larval counts were higher (0.06-17.8 larvae per gram of feces). This work provides the first experimental completion of the life-cycle of V. eleguneniensis.

Bionomics of larvae of Parelaphostrongylus odocoilei (Nematoda: Protostrongylidae) in experimentally infected gastropod intermediate hosts.

Parelaphostrongylus odocoilei is a protostrongylid parasite that has recently been recognized at several locations in sub-Arctic, but not Arctic, North America. We investigated factors that may determine the distribution of P. odocoilei, including suitable gastropod intermediate hosts, temperature requirements for larval development in gastropods, and larval emergence facilitating overwinter transmission. We collected and experimentally infected gastropods from a site in the sub-Arctic where P. odocoilei is at the northern limit of its distribution. Deroceras laeve, Catinella sp., and Euconulus cf fulvus, but not members of the Pupillidae, were suitable intermediate hosts. We describe bionomics of larvae of P. odocoilei in D. laeve and Catinella sp. Infective larvae emerged from all slugs (D. laeve) and 60% of Catinella sp. snails, and emergence from D. laeve was intensity dependent. Emerged infective larvae survived up to 6 mo under conditions approximating that of the subnivean environment. In D. laeve, there was a direct relationship between temperature and development rate of larvae of P. odocoilei. Larvae of P. odocoilei did not develop to infective stage below the theoretical threshold (8.5 C), and required a minimum of 163 degree days to complete development. These developmental parameters can be incorporated into a model to predict larval development in the field. Knowledge of the factors influencing larval bionomics provides the foundation for predicting temporal and spatial patterns of parasite distribution, abundance, and transmission.

Parasites, stress and reindeer: infection with abomasal nematodes is not associated with elevated glucocorticoid levels in hair or faeces.

Stress hormones (glucocorticoids), incorporated into hair/fur and faeces, have been proposed as biomarkers of overall health in wildlife. Although such biomarkers may be helpful for wildlife conservation and management, their use has rarely been validated. There is a paucity of studies examining the variation of stress hormones in mammals and how they relate to other health measures, such as parasitism. Parasites are ubiquitous in wildlife and can influence the fitness of individual animals and populations. Through a longitudinal experiment using captive reindeer (Rangifer tarandus tarandus), we tested whether animals infected with Ostertagia gruehneri, a gastrointestinal nematode with negative impacts on fitness of the host, had higher stress levels compared with those that had been treated to remove infection. Faecal samples were collected weekly for 12 weeks (June-September) and hair was collected at the start and end of the study; glucocorticoids were quantified using enzyme immunoassays. Contrary to what was expected, infected reindeer had similar levels of cortisol in hair and slightly lower glucocorticoid metabolites in faeces compared with uninfected reindeer. Faecal corticosterone levels were higher than faecal cortisol levels, and only corticosterone increased significantly after a handling event. These results suggest that reindeer may use a tolerance strategy to cope with gastrointestinal nematodes and raise the question as to whether moderate infection intensities with nematodes are beneficial to the host. By removing nematodes we may have altered the gut microbiota, leading to the observed elevated faecal glucocorticoid metabolite levels in the treated reindeer. These findings demonstrate the importance of considering both cortisol and corticosterone in physiological studies, as there is mounting evidence that they may have different functionalities.

Global warming is changing the dynamics of Arctic host-parasite systems.

Global climate change is altering the ecology of infectious agents and driving the emergence of disease in people, domestic animals, and wildlife. We present a novel, empirically based, predictive model for the impact of climate warming on development rates and availability of an important parasitic nematode of muskoxen in the Canadian Arctic, a region that is particularly vulnerable to climate change. Using this model, we show that warming in the Arctic may have already radically altered the transmission dynamics of this parasite, escalating infection pressure for muskoxen, and that this trend is expected to continue. This work establishes a foundation for understanding responses to climate change of other host-parasite systems, in the Arctic and globally.

A new lungworm in muskoxen: an exploration in Arctic parasitology.

Ruminants are vital elements of the Holarctic ecosystem. Little is known, however, of the structure or biology of their parasite fauna, particularly in North America. Global warming, coupled with increasing human activity in the Arctic, requires enhanced international interdisciplinary efforts to better understand the many factors, including parasites, that influence the population health of caribou, reindeer, muskoxen and wild sheep. The discovery of an unusual new genus of protostrongylid lung nematode in muskoxen from the central Canadian Arctic is described, and the intricacies of the parasite's relationship with its muskoxen definitive hosts, its gastropod intermediate hosts and the arctic environment are discussed.

Hepatic gene expression patterns in thyroid hormone-treated hypothyroid rats.

Thyroid hormone (T3) is essential for normal development, differentiation and metabolic balance. We have performed DNA microarray experiments using hepatic RNA from hypothyroid and T3-treated hypothyroid rats in order to characterize T3-induced gene expression patterns after various time points (6, 24 and 48 h after the administration of the hormone). Sixty-two of 4608 different genes displayed a reproducible T3-response, and cluster analysis divided these differentially regulated genes into six expression patterns. Thirty-six genes were not significantly regulated within the first 24 h. Transient transfection experiments of eight late-induced gene promoters failed to detect a thyroid hormone response element within their regulatory elements, suggesting an indirect activation mechanism(s). In search for an intermediate factor of T3 action, we examined whether various rather ubiquitous transcription factors, peroxisome proliferator-activated receptors (PPARs) and coactivators of the PPARgamma coactivator 1 family (PGC-1) are regulated by T3. Only PPARgamma and PERC/PGC-1beta exhibit a significant T3-response within the first 6 h after treatment, identifying these factors as candidate components for mediating the late-induced expression pattern. Regulation of early-induced genes within the first 6 h after administration of T3 on transcript levels correlates with altered protein levels after 24 and 48 h in vivo.

Immune mechanisms in byssinosis.

The pathogenesis of byssinosis has been attributed to several different immunopathologic mechanisms, including a type III (immune complex) pulmonary injury. To further examine this type III theory, sera (Monday preshift) from 59 cotton textile workers were examined by gel diffusion and counterimmunoelectrophoresis for precipitating antibodies to aqueous extracts of cotton bract, carpels, stems, leaves, immature cotton bract, carpels, stems, leaves, immature cotton lint, and cardroom cotton dust. Sera were also collected from 35 nonexposed normal volunteers and examined similarly. No true precipitating antibodies to these extracts could be detected in any of the control or worker serum samples. The aqueous extracts of cardroom cotton dust and cotton stems were found to contain naturally occurring components that precipitated (in agarose gel) beta-lipoprotein and gamma-globulins (mostly IgG) in a nonimmunologic manner. Sera from normal human controls and cotton textile workers all produced identical patterns of reaction with these two extracts. Treatment of these extracts with polyvinylpolypyrrolidone, a specific insoluble adsorbent for polyphenolic tannins, eliminated this pseudoimmune reaction. Although the role this pseudoimmune reaction may play in the pathogenesis of byssinosis is still unknown, it demonstrates the problems associated with laboratory-based investigations of the immunologic aspects of byssinosis.

Temporal profile of release of neurobiochemical markers of brain damage after traumatic brain injury is associated with intracranial pathology as demonstrated in cranial computerized tomography.

This study aimed at the investigation of release patterns of neuron specific enolase (NSE) and protein S-100B after traumatic brain injury (TBI) and their association with intracranial pathologic changes as demonstrated in computerized tomography (CT). We analyzed NSE and S-100B concentrations in serial venous blood samples taken one to three days after TBI in 66 patients by the use of immunoluminometric assays. These markers are considered to be specific neurobiochemical indicators of damage to glial (S-100B) or neuronal (NSE) brain tissue. Standardized neurological examination and plani- and volumetric evaluation of computerized tomography scans were performed in all patients. Patients with medium severe to severe TBI [Glasgow Coma Scale (GCS) score at the site of accident < or =12] exhibited significantly higher NSE and S-100B concentrations and a significantly longer release compared to patients with minor head injury (GCS: 13-15). Both, patients with and without visible intracerebral pathology in CT scans exhibited elevated concentrations of NSE and S-100B after TBI and a significant decrease in the follow-up blood samples. Release patterns of S-100B and NSE differed in patients with primary cortical contusions, diffuse axonal injury (DAI), and signs of cerebral edema (ICP) without focal mass lesions. All serum concentrations of NSE and S-100B were significantly correlated with the volume of contusions. The data of the present study indicate that the early release patterns of NSE and S-100 may mirror different pathophysiological consequences of traumatic brain injury.

Atheromas of the thoracic aorta: A comparison of transesophageal echocardiography and breath-hold gadolinium-enhanced 3-dimensional magnetic resonance angiography.

Transesophageal echocardiography (TEE) has been the procedure of choice for identifying thoracic aortic atheromas. All patients over a 2-year period who underwent both TEE and magnetic resonance angiography of the thoracic aorta within 1 month were identified. The largest plaque in 3 aortic segments (ascending, arch, and descending) was measured. Thirty patients (16 men; 66.8 +/- 12.9 years) were studied. The mean size of the atheromas in the arch was larger as measured by TEE compared with magnetic resonance angiography (3.4 vs 1.4 mm, P =.01). However, the mean atheroma size was similar in the ascending aorta (1.9 vs 1.3 mm, P =.5) and descending aorta (3.9 vs 3.5 mm, P =.66). Of 24 aortic segments with plaques measuring >/=5 mm (with high embolic risk), 22 (92%) were seen on TEE and only 13 (54%) on magnetic resonance angiography (P =. 003). In conclusion, although both techniques are complementary, TEE does identify more high-risk plaques.

Modified malis bayonet forceps aids application of the cyberonics vagus nerve stimulator electrode: technical note.

OBJECTIVE: To notify neurosurgeons about a modified bayonet forceps that aids application of the vagus nerve stimulating electrode. METHODS: The manufacturer (Codman & Shurtleff, Inc., Raynham, MA) extended the tips of an upward-angled Malis bayonet forceps from 2 mm to 6 mm. RESULTS: The modified bayonet tips, when placed under the vagus nerve, extend well beyond the edge of the usual vagus nerve to easily accept the electrode lead. CONCLUSION: The modified bayonet forceps and depicted wrapping sequence shorten electrode wrapping time.

Emergence of third-stage larvae of Umingmakstrongylus pallikuukensis from three gastropod intermediate host species.

We investigated the emergence of third-stage larvae (L3) of Umingmakstrongylus pallikuukensis from the slugs Deroceras laeve, Deroceras reticulatum, and the snail Catinella sp. in the laboratory and from D. laeve on the tundra. Third-stage larvae emerged from 8 of 8 D. laeve and 8 of 8 D. reticulatum housed at 20 C in darkness and from 9 of 10 D. laeve and 5 of 5 Catinella sp. housed at 21 C with 10-12 hr of light/day. Larvae emerged from D. laeve and D. reticulatum over a wide range of infection intensities (2-179 and 20-65, respectively), and the patterns of emergence were independent of intensity. The majority of the L3 emerged from most of the Deroceras spp. by 58 or 60 days postinfection (PI). Lower rates of emergence were observed from Catinella sp. Larvae emerged from D. laeve on the tundra by 10 wk PI and were recovered from the vegetation in some experimental enclosures the following year. Third-stage larvae survived in tap and distilled water at 0-4 C for 13 mo. Emergence of L3 of U. pallikuukensis from the intermediate host may increase the temporal and spatial availability of L3 and enhance their survival and transmission.

Umingmakstrongylus pallikuukensis (Nematoda: Protostrongylidae) in gastropods: larval morphology, morphometrics, and development rates.

Morphological and morphometric aspects of larval development of Umingmakstrongylus pallikuukensis in Deroceras laeve and the effects of temperature on development rates in D. laeve and Deroceras reticulatum were investigated in the laboratory. Larval stages were best differentiated by separation of cuticular sheaths, tail structure, and viability following digestion. Growth in body and esophagus width was observed during the first-stage within the intermediate host, but the major increases in body length and width occurred immediately following the second molt. Larval development in D. laeve and D. reticulatum occurred more rapidly at warmer temperatures. The calculated threshold temperatures were 8.5 and 9.5 C in D. laeve and D. reticulatum, respectively, and 167 degree-days were required for development to third-stage larvae (L3) in both hosts. These thresholds are higher than those calculated from published data for the closely related Muellerius capillaris (4.2 C) but are similar to those for the more distantly related northern protostrongylid, Elaphostrongylus rangiferi (8.3-10.3 C). Conversely, degree-days required for development to infective L3 were more similar among the Muelleriinae than between this group and the Elaphostrongylinae. Developmental parameters for protostrongylid larvae may be influenced both by the environment and by features of the parasites and the intermediate hosts, including phylogeny.