Douglas P. Whiteside

Dogs as Sources and Sentinels of Parasites in Humans and Wildlife, Northern Canada

A minimum of 11 genera of parasites, including 7 known or suspected to cause zoonoses, were detected in dogs in 2 northern Canadian communities. Dogs in remote settlements receive minimal veterinary care and may serve as sources and sentinels for parasites in persons and wildlife, and as parasite bridges between wildlife and humans.

Concentrations of 17 elements, including mercury, and their relationship to fitness measures in arctic shorebirds and their eggs.

Exposure to contaminants is one hypothesis proposed to explain the global decline in shorebirds, and this is of particular concern in the arctic. However, little information exists on contaminant levels in arctic-breeding shorebirds, especially in Canada. We studied potential contaminants in three biparental shorebird species nesting in Nunavut, Canada: ruddy turnstones (Arenaria interpres), black-bellied plovers (Pluvialis squatarola) and semipalmated plovers (Charadrius semipalmatus). Blood, feathers and eggs were analyzed for As, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Tl, V, and Zn. We assessed whether element concentrations a) differed among species and sexes, b) were correlated among pairs and their eggs, and c) were related to fitness endpoints, namely body condition, blood-parasite load, nest survival days, and hatching success. Non-essential elements were found at lower concentrations than essential elements, with the exception of Hg. Maximum Hg levels in blood approached those associated with toxicological effects in other bird species, but other elements were well below known toxicological thresholds. Reproductive success was negatively correlated with paternal Hg and maternal Pb, although these effects were generally weak and varied among tissues. Element levels were positively correlated within pairs for blood-Hg (turnstones) and feather-Ni and Cr (semipalmated plovers); concentrations in eggs and maternal blood were never correlated. Concentrations of many elements differed among species, but there was no evidence that any species had higher overall exposure to non-essential metals. In conclusion, whereas we found little evidence that exposure to the majority of these elements is leading to declines of the species studied here, Hg levels were of potential concern and both Hg and Pb warrant further monitoring.

Cortisol and corticosterone independence in cortisol-dominant wildlife

Species have traditionally been defined as cortisol-dominant or corticosterone-dominant, depending on the glucocorticoid that is reported. To assess the degree of covariance versus independence between cortisol and corticosterone, 245 serum samples belonging to 219 individuals from 18 cortisol-dominant, non-domesticated species (6 mammalian orders) were compared by mass spectrometry. In these samples, which were elevated above baseline, concentration ranges were overlapping for cortisol and corticosterone although cortisol was dominant in every sample except one of 17 bighorn sheep with a corticosterone-biased cortisol-to-corticosterone ratio of 0.17. As expected, cortisol and corticosterone were strongly associated among species (r(2)=0.8; species with high absolute cortisol tend to have high absolute corticosterone concentrations), with wide variation in the species-average cortisol-to-corticosterone ratio (range 7.5-49) and an even wider ratio range across individuals (0.2-341). However, only 9 out of 13 species with >7 individuals showed a positive association between cortisol and corticosterone among individuals, and repeated measures of the cortisol-to-corticosterone ratio within individuals were weakly associated (CV range 3-136%). We conclude that corticosterone, although at lower concentrations, has the potential to signal independently of cortisol, and should be included in integrated endocrine models of stress responses.

Concentrations of 17 elements, including mercury, in the tissues, food and abiotic environment of Arctic shorebirds

Exposure to contaminants is one hypothesis proposed to explain the global decline in shorebirds, and is also an increasing concern in the Arctic. We assessed potential contaminants (As, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Tl, V, and Zn) at a shorebird breeding site in Nunavut, Canada. We compared element levels in soil, invertebrates and shorebird blood to assess evidence for bioconcentration and biomagnification within the Arctic-based food chain. We tested whether elements in blood, feathers and eggs of six shorebird species (Pluvialis squatarola, Calidris alpina, C. fuscicollis, Phalaropus fulicarius, Charadrius semipalmatus, and Arenaria interpres) were related to fitness endpoints: adult body condition, blood-parasite load, egg size, eggshell thickness, nest duration, and hatching success. To facilitate comparison to other sites, we summarise the published data on toxic metals in shorebird blood and egg contents. Element concentrations and invertebrate composition differed strongly among habitats, and habitat use and element concentrations differed among shorebird species. Hg, Se, Cd, Cu, and Zn bioconcentrated from soil to invertebrates, and Hg, Se and Fe biomagnified from invertebrates to shorebird blood. As, Ni, Pb, Co and Mn showed significant biodilution from soil to invertebrates to shorebirds. Soil element levels were within Canadian guidelines, and invertebrate Hg levels were below dietary levels suggested for the protection of wildlife. However, maximum Hg in blood and eggs approached levels associated with toxicological effects and Hg-pollution in other bird species. Parental blood-Hg was negatively related to egg volume, although the relationship varied among species. No other elements approached established toxicological thresholds. In conclusion, whereas we found little evidence that exposure to elements at this site is leading to the declines of the species studied, Hg, as found elsewhere in the Canadian Arctic, is of potential concern for breeding bird populations.

Evaluation of an animal protein-free semen extender for cryopreservation of epididymal sperm from North American bison (Bison bison).

The objective was to evaluate the suitability of an animal protein-free semen extender for cryopreservation of epididymal sperm from the two subspecies of North American bison: plains (Bison bison bison) and wood (Bison bison athabascae) bison. Both cauda epididymides (from six plains and five wood bison) were minced and incubated in Sp-TALPH buffer for approximately 2 h at 37 °C to release actively motile sperm. Sperm suspensions were filtered, centrifuged and the sperm pellet from each bull was divided into two fractions and diluted either in egg yolk containing extender, Triladyl, or in an animal protein-free extender, Andromed, and equilibrated for 20 min at 37 °C. Thereafter, samples were chilled and cryopreserved. Frozen-thawed sperm were evaluated for motility (computer assisted sperm analysis), viability (SYBR 14 and propidium iodide), acrosome integrity (FITC conjugated PSA), cryocapacitation (tyrosine phosphorylation of sperm proteins as a biomarker), and fertilizing ability (in a heterologous IVF system). There was no significant difference for progressive motility, viability, and acrosome integrity between the two extenders for plains bison (36.8 ± 9.0, 60.5 ± 17.4, and 77.3 ± 4.6%; overall mean ± SD) as well as for wood bison (11.7 ± 8.1, 13.7 ± 5.6, and 73.4 ± 4.2%). Levels of tyrosine phosphorylation did not differ for sperm preserved in the two extenders for both subspecies, although an inter-bull variability in the response to tyrosine phosphorylation between extenders was suggested for plains bison. Fertilization percent did not differ significantly between extenders for plains bison (84.16 ± 9.92%, overall mean ± SD) and for wood bison (59.53 ± 19.99%). In conclusion, in the absence of significant difference between extenders in post-thaw sperm characteristics, we inferred that Andromed (animal protein-free) was suitable for cryopreservation of epididymal sperm from North American bison.

Evaluation of the enteric microflora of captive whooping cranes (Grus americana) and sandhill cranes (Grus canadensis).

The enteric flora of captive whooping cranes (Grus americana) and sandhill cranes (Grus canadensis) has not been well described, despite its potential importance in the understanding of both the normal condition of the intestinal physiology of these animals and the altered colonization within disease states in these birds. Nineteen whooping cranes and 23 sandhill cranes housed currently at the Calgary Zoo or its affiliated Devonian Wildlife Conservation Centre (DWCC) in Calgary, Alberta were sampled from October 2004-February 2005 by collecting aerobic and anaerobic cloacal swabs from each bird. There were seven major groupings of bacteria isolated from both species of crane. Gram-positive cocci, coliforms, and gram-negative bacilli were the most prevalent types of bacteria isolated for both crane species, with Escherichia coli, Enterococcus faecalis, and Streptococcus Group D, not Enterococcus the bacterial species isolated most commonly. There was a significant difference in the average number of isolates per individual between the two crane species but no differences between age or gender categories within crane species. Campylobacter sp. were isolated from five whooping cranes. The potential zoonotic pathogen Campylobacter jejuni was isolated from one whooping crane and C. upsaliensis was isolated from a second. Three other isolates were unspeciated members of the Campylobacter genus and likely belong to a species undescribed previously. The evaluation of the enteric cloacal flora of whooping cranes and sandhill cranes illustrates that differences exist between these two closely related crane species, and highlights the potential implications these differences may have for current practices involving captive wildlife. Zoo Biol 0:1-13, 2007. (c) 2007 Wiley-Liss, Inc.

Recovery and cryopreservation of epididymal sperm of plains bison (Bison bison bison) as a model for salvaging the genetics of wood bison (Bison bison athabascae).

The objective of this study was to optimize recovery and cryopreservation of epididymal sperm from plains bison, as a model for wood bison. In Phase 1, cauda epididymides were recovered from bison (n = 14) immediately after slaughter, minced and incubated in Sp-TALPH buffer for 3 h at 36 degrees C. The resulting sperm suspensions were cryopreserved in Triladyl, using a protocol for bovine semen. In Phase 2, epididymal sperm were cryopreserved in either Triladyl or Andromed. The mean (+/-SD) estimated number of sperm recovered was 468 +/- 207 x 10(6). There was an increase (p < 0.05) in the proportion of sperm with normal morphology between initial recovery and after extension (52.4 +/- 4.6 vs 69.7 +/- 2.4%), with a concurrent decrease (p < 0.05) in the proportion of sperm with distal droplets. Median values for progressively motile sperm in post-thaw samples (60%) were lower (p < 0.05) than that after extension or after chilling (70% for both). The mean percentages of viable sperm and of sperm with an intact acrosome were lower (p < 0.05) for frozen-thawed samples (38.7 +/- 2.8 and 85.2 +/- 1.1) compared with extended (66.2 +/- 2.2 and 92.4 +/- 0.9) or chilled (63.7 +/- 2.5 and 90.0 +/- 1.0) samples. Rates of cleavage, morulae and blastocyst production were not significantly different for chilled (70.9, 38.7 and 8.0%) vs post-thaw sperm (73.0, 46.0 and 6.3%). There was no significant difference between extenders for most sperm characteristics. In conclusion, we developed a functional protocol for the recovery and cryopreservation of epididymal sperm from plains bison, which may have implications for the genetic preservation of wood bison.

Sperm characteristics in plains (Bison bison bison) versus wood (Bison bison athabascae) bison.

The objective was to compare sperm characteristics between the two subspecies of North American bison, plains bison (Bison bison bison) and wood bison (Bison bison athabascae). Frozen-thawed ejaculated sperm from age-matched plains (n = 3) and wood (n = 2) bison were evaluated for morphometry, motility, viability, protein profile, and in vitro fertilization characteristics. Sperm morphometry and motility were assessed with computer-based systems, viability was assessed with SYBR-14 and propidium iodide, and fertilizing ability was determined using a heterologous in vitro fertilization system (using bovine oocytes). For plains versus wood bison, there were significant differences for head width (4.76 ± 0.22 vs 4.71 ± 0.19 μm; mean ± SD), head area (35.64 ± 1.91 vs 34.72 ± 2.64 μm(2)), head perimeter (23.61 ± 0.68 vs 23.31 ± 0.98 μm), midpiece length (14.58 ± 0.4 vs 14.36 ± 0.51 μm), midpiece width (0.81 ± 0.06 vs 0.79 ± 0.07 μm), and tail length (46.61 ± 2.15 vs 45.98 ± 2.08 μm). However, there was no significant difference in head length (overall, 9.04 ± 0.37 μm), progressive motility (41.16 ± 8.39%), or viability (41.58 ± 5.58%). Based on two-dimensional gel electrophoresis, 93 out of 113 protein spots were similar in their expression patterns. Furthermore, we inferred that differences in sperm biometry between these subspecies did not affect in vitro fertilization percentage (overall, 82.62 ± 12.13%). Based on these findings, we concluded that plains bison were an appropriate research model for developing reproductive technologies for wood bison.

MELOXICAM AND SURGICAL DENERVATION OF THE COXOFEMORAL JOINT FOR THE TREATMENT OF DEGENERATIVE OSTEOARTHRITIS IN A BENGAL TIGER (PANTHERA TIGRIS TIGRIS)

Abstract An adult male white Bengal tiger (Panthera tigris tigris) with pronounced atrophy of the pelvic musculature was diagnosed with degenerative osteoarthritis of the coxofemoral joints. Initial management with the nonsteroidal anti-inflammatory drug meloxicam and a semisynthetic sodium pentosan polysulfate resulted in clinical improvement and radiographic stabilization of the arthritic condition over several months. However, because pain was still evident, bilateral denervation of the coxofemoral joints was performed, successfully ameliorating the signs of osteoarthritic pain in the tiger. Meloxicam has shown good clinical efficacy for the treatment of osteoarthritis and other painful conditions in large felids. Coxofemoral joint denervation offers many advantages for the treatment of osteoarthritis in exotic carnivore species, and should be considered a viable treatment modality.

Effect of Reproductive Seasonality on Gamete Quality in the North American Bison (Bison bison bison)

The objective was to investigate the effects of reproductive seasonality on gamete quality in plains bison (Bison bison bison). Epididymal sperm (n = 61 per season), collected during the breeding season (July-September), had significantly higher post-thaw total motility (36.76 ± 14.18 vs 31.24 ± 12.74%), and lower linearity (0.36 ± 0.06 vs 0.39 ± 0.04) and wobbliness (0.49 ± 0.04 vs 0.51 ± 0.03; mean ± SD) compared to non-breeding season (January-March) samples. Representative samples (n = 4) from each season were used in heterologous IVF trials using cattle oocytes. Cleavage, morulae and blastocyst percentage were higher for breeding vs non-breeding season sperm samples (81.88 ± 6.8 vs 49.94 ± 6.77; 41.89 ± 13.40 vs 27.08 ± 23.21; and 30.49 ± 17.87 vs 13.72 ± 18.98%, respectively). Plains bison ovaries collected during the breeding (n = 97 pairs) and non-breeding (n = 100 pairs) seasons were classified as luteal or follicular. Oocytes recovered from these ovaries were classified into five grades based on morphology. There was no significant difference in the number of luteal ovaries or grades of oocytes recovered. Oocytes were matured, fertilized (with frozen sperm from three bison bulls) and cultured in vitro. Cleavage percentage was higher for oocytes collected during breeding vs non-breeding season (83.72 ± 6.42 vs 73.98 ± 6.43), with no significant difference in subsequent development to blastocysts. In summary, epididymal sperm from non-breeding season had decreased total motility and resulted in reduced embryo production in vitro. Oocytes collected during non-breeding season had reduced ability to be matured, fertilized and/or undergo cleavage in vitro. Data suggested that season influenced gamete quality in plains bison.