VALIDATION OF ENZYME-LINKED IMMUNOSORBENT ASSAY TECHNIQUES TO MEASURE SERUM DEHYDROEPIANDROSTERONE (DHEA) AND DHEA-S IN NARWHALS (MONODON MONOCEROS).

Narwhals (Monodon monoceros) are increasingly exposed to anthropogenic disturbances that may increase their stress levels with unknown consequences for the overall population dynamics. The validation and measurement of chronic stress biomarkers could contribute toward improved understanding and conservation efforts for this species. Dehydroepiandrosterone (DHEA) and its sulfated metabolite DHEA-S are collectively referred to as DHEA(S). Serum DHEA(S) concentrations combined in ratios with cortisol [cortisol/DHEA(S)] have been shown to be promising indicators of chronic stress in humans, domestic animals, and wildlife. During field tagging in 2017 and 2018 in Baffin Bay, Nunavut, Canada, 14 wild narwhals were sampled at the beginning and end of the capture-tagging procedures. Serum DHEA(S) were measured with commercially available competitive enzyme-linked immunosorbent assays (ELISA) developed for humans. A partial validation of the ELISA assays was performed by the determination of the intra-assay coefficient of variation, confirmation of the DHEA(S) dilutional linearity, and the calculation of the percentage of recovery. Mean values (nanograms per milliliter ± standard error of the mean) of narwhal serum cortisol, DHEA(S), and cortisol/DHEA(S) ratios, at the beginning and at the end of handling, respectively, are reported (cortisol = 30.74 ± 4.87 and 41.83 ± 4.83; DHEA = 1.01 ± 0.52 and 0.99 ± 0.50; DHEA-S = 8.72 ± 1.68 and 7.70 ± 1.02; cortisol/DHEA = 75.43 ± 24.35 and 84.41 ± 11.76, and cortisol/DHEA-S = 4.16 ± 1.07 and 6.14 ± 1.00). Serum cortisol and cortisol/DHEA-S were statistically higher at the end of the capture (P= 0.024 and P= 0.035, respectively). Moreover, serum cortisol at the end of handling was positively correlated to total body length (P = 0.042) and tended to be higher in males (P = 0.086). These assays proved easy to perform, rapid, and suitable for measuring serum DHEA(S) of narwhals and that calculated cortisol/DHEA(S) are potential biomarkers for chronic stress in narwhals and possibly other cetaceans.

ASSESSING THE HUMORAL RESPONSE TO AND SAFETY OF A COMMERCIALLY AVAILABLE EQUINE WEST NILE VIRUS VACCINE IN A ZOO-BASED CONSERVATION BREEDING POPULATION OF ENDANGERED GREATER SAGE-GROUSE (CENTROCERCUS UROPHASIANUS).

Greater sage-grouse (Centrocercus urophasianus) are an endangered species in Canada and have been extirpated from five states in the United States. Infectious diseases can pose a challenge to conservation efforts, and the greater sage-grouse is susceptible to West Nile virus (WNV). This study measured the humoral response to a commercially available WNV vaccine in a zoo-based conservation breeding population of greater sage-grouse. Since the initiation of the conservation program in 2014, all sage-grouse at the Calgary Zoo's Wildlife Conservation Centre have been vaccinated against WNV. Juveniles received a series of three vaccines every 2-3 wk starting around 2 wk of age; adults had received the same vaccination series at hatch and had since been boostered annually. Antibody titers were measured on 60 serum samples from 36 juveniles and 18 adults collected in 2018 using either serum neutralization or plaque-reduction neutralization methods. No detectable antibody response was noted in juveniles after their second (n = 17) or third booster vaccinations (n = 23). Only 35% of 20 adult samples collected had positive titers even after multiple years of vaccination. Only two cases of adverse vaccine reaction have been noted in this species with over 800 doses being administered between 2014 and 2020. Despite a lack of antibody response in juveniles and most adults, there have been no WNV-associated deaths in this population even with confirmed cases of WNV in free-ranging greater sage-grouse and other species in the area during this time frame, suggesting that cell-mediated immunity may be of greater importance for protection against WNV infections in this species. The initial vaccination series was changed in 2019 to a series of only two doses of vaccine, and no clinical cases or mortality from WNV occurred in 2019 or 2020 with the new protocol.

Variation in the hemostatic complement (C5a) responses to in vitro nitrogen bubbles in monodontids and phocids.

Immune responses to nitrogen gas bubbles, particularly activation of inflammation via the complement cascade, have been linked to the development of symptoms and damage associated with decompression sickness (DCS) in humans. Marine mammals were long thought not to be susceptible to such dive-related injury, yet evidence of DCS-like injury and new models of tissue nitrogen super-saturation suggest that bubbles may routinely form. As such, it is possible that marine mammals have protective adaptations that allow them to deal with a certain level of bubble formation during normal dives, without acute adverse effects. This work evaluated the complement response, indicative of inflammation, to in vitro nitrogen bubble exposures in several marine mammal species to assess whether a less-responsive immune system serves a protective role against DCS-like injury in these animals. Serum samples from beluga (Delphinapterus leucas), and harbor seals (Phoca vitulina) (relatively shallow divers) and deep diving narwhal (Monodon monoceros), and Weddell seals (Leptonychotes weddellii) were exposed to nitrogen bubbles in vitro. Complement activity was evaluated by measuring changes in the terminal protein C5a in serum, and results suggest marine mammal complement is less sensitive to gas bubbles than human complement, but the response varies between species. Species-specific differences may be related to dive ability, and suggest moderate or shallow divers may be more susceptible to DCS-like injury. This information is an important consideration in assessing the impact of changing dive behaviors in response to anthropogenic stressors, startle responses, or changing environmental conditions that affect prey depth distributions.

Actual versus perceived peer sexual risk behavior in online youth social networks.

Perception of peer behaviors is an important predictor of actual risk behaviors among youth. However, we lack understanding of peer influence through social media and of actual and perceived peer behavior concordance. The purpose of this research is to document the relationship between individual perception of and actual peer sexual risk behavior using online social networks. The data are a result of a secondary analysis of baseline self-reported and peer-reported sexual risk behavior from a cluster randomized trial including 1,029 persons from 162 virtual networks. Individuals (seeds) recruited up to three friends who then recruited additional friends, extending three waves from the seed. ANOVA models compared network means of actual participant behavior across categories of perceived behavior. Concordance varied between reported and perceived behavior, with higher concordance between perceived and reported condom use, multiple partners, concurrent partners, sexual pressure, and drug and alcohol use during sex. Individuals significantly over-reported risk and under-reported protective peer behaviors related to sex.

Social media-delivered sexual health intervention: a cluster randomized controlled trial.

BACKGROUND: Youth are using social media regularly and represent a group facing substantial risk for sexually transmitted infection (STI). Although there is evidence that the Internet can be used effectively in supporting healthy sexual behavior, this has not yet extended to social networking sites. PURPOSE: To determine whether STI prevention messages delivered via Facebook are efficacious in preventing increases in sexual risk behavior at 2 and 6 months. DESIGN: Cluster RCT, October 2010-May 2011. SETTING/PARTICIPANTS: Individuals (seeds) recruited in multiple settings (online, via newspaper ads and face-to-face) were asked to recruit three friends, who in turn recruited additional friends, extending three waves from the seed. Seeds and waves of friends were considered networks and exposed to either the intervention or control condition. INTERVENTION: Exposure to Just/Us, a Facebook page developed with youth input, or to control content on 18-24 News, a Facebook page with current events for 2 months. MAIN OUTCOME MEASURES: Condom use at last sex and proportion of sex acts protected by condoms. Repeated measures of nested data were used to model main effects of exposure to Just/Us and time by treatment interaction. RESULTS: A total of 1578 participants enrolled, with 14% Latino and 35% African-American; 75% of participants completed at least one study follow-up. Time by treatment effects were observed at 2 months for condom use (intervention 68% vs control 56%, p=0.04) and proportion of sex acts protected by condoms (intervention 63% vs control 57%, p=0.03) where intervention participation reduced the tendency for condom use to decrease over time. No effects were seen at 6 months. CONCLUSIONS: Social networking sites may be venues for efficacious health education interventions. More work is needed to understand what elements of social media are compelling, how network membership influences effects, and whether linking social media to clinical and social services can be beneficial. TRIAL REGISTRATION: This study is registered at www.clinicaltrials.govNCT00725959.

Conservative management of pyothorax in an Amur tiger (Panthera tigris altaica).

Therapy for pyothorax, or pleural empyema, has not been described for large felids. This case describes the successful treatment of pyothorax in a captive, large felid. A 15-yr-old multiparous, female Amur tiger (Panthera tigris altaica) presented with nonspecific clinical signs caused by an insidious onset of pyothorax. Management of pyothorax cases in companion animals often involves thoracostomy tube placement with recurrent drainage of the pleural cavity, intensive supportive care, and monitoring. In this case, conservative management was elected because hospitalization was not a viable option. Thoracocentesis was performed to drain as much fluid from the chest cavity as possible, yielding more than 1.3 L. Corynebacterium sp. and unidentified anaerobic gram-positive cocci were cultured. Treatment included cefovecin subcutaneously, oral antibiotic therapy with clindamycin and marbofloxacin, meloxicam, and restricted exercise by minimizing access to the main exhibit. Significant improvement was noted clinically and radiographically 6 wk later, and no relapses were noted in the following weeks. An examination 11.5 mo later confirmed resolution.

Case study: An ethics case study of HIV prevention research on Facebook: the Just/Us study.

OBJECTIVE: To consider issues related to research with youth on social networking sites online. METHODS: Description of the data collection process from 1,588 participants in a randomized controlled trial testing the efficacy of HIV prevention education delivered on Facebook. Using respondent-driven sampling, staff-recruited participants are encouraged to recruit up to three friends to enroll in the study. RESULTS: Researchers should (a) consider whether an online social networking site is an appropriate place to implement a research study; (b) offer opportunities to review informed consent documents at multiple times and in multiple locations throughout the study; and (c) collect data outside the social networking site and store it behind secure firewalls to ensure it will not be accessible to any person on the social networking site. CONCLUSIONS: Online social networks are growing in popularity. Conducting research on social media sites requires deliberate attention to consent, confidentiality, and security.

Nasal osteosarcoma and interstitial cell tumor in a Vancouver Island marmot (Marmota vancouverensis).

A 6-yr-old male Vancouver Island marmot (Marmota vancouverensis) presented for poor hibernation, weight loss, and symmetric trunk alopecia. An abdominal interstitial cell tumor was identified and surgically removed. Serum levels of estrogen were markedly elevated before surgery and decreased after tumor removal, indicating that the tumor had been functionally secretory. Nine months later, the marmot presented with respiratory stridor. A large boney nasal mass was identified radiographically and evaluated by computed tomography (CT) prior to surgical debulking. The marmot did not recover from anesthesia. Pathologic findings included a nasal osteosarcoma with lysis of the cribriform plate, and endocardial fibrosis with degenerative changes within the adjoining myocardium. This is the first known report of nasal osteosarcoma and interstitial tumor in a Vancouver Island marmot.

Human papillomavirus vaccination practices: a survey of US physicians 18 months after licensure.

OBJECTIVES: The objectives of this study were to assess, in a nationally representative network of pediatricians and family physicians, (1) human papillomavirus (HPV) vaccination practices, (2) perceived barriers to vaccination, and (3) factors associated with whether physicians strongly recommended HPV vaccine to 11- to 12-year-old female patients. METHODS: In January through March 2008, a survey was administered to 429 pediatricians and 419 family physicians. RESULTS: Response rates were 81% for pediatricians and 79% for family physicians. Ninety-eight percent of pediatricians and 88% of family physicians were administering HPV vaccine in their offices (P<.001). Among those physicians, fewer strongly recommended HPV vaccination for 11- to 12-year-old female patients than for older female patients (pediatricians: 57% for 11- to 12-year-old patients and 90% for 13- to 15-year-old patients; P<.001; family physicians: 50% and 86%, respectively; P<.001). The most-frequently reported barriers to HPV vaccination were financial, including vaccine costs and insurance coverage. Factors associated with not strongly recommending HPV vaccine to 11- to 12-year-old female patients included considering it necessary to discuss sexuality before recommending HPV vaccine (risk ratio: 1.27 [95% confidence interval: 1.07-1.51]) and reporting more vaccine refusals among parents of younger versus older adolescents (risk ratio: 2.09 [95% confidence interval: 1.66-2.81]). CONCLUSIONS: Eighteen months after licensure, the vast majority of pediatricians and family physicians reported offering HPV vaccine. Fewer physicians strongly recommended the vaccine for younger adolescents than for older adolescents, and physicians reported financial obstacles to vaccination.

Knowledge of interim recommendations and use of Hib vaccine during vaccine shortages.

OBJECTIVES: The goals were to determine among pediatricians and family physicians (1) knowledge of interim recommendations regarding Haemophilus influenzae type b (Hib) vaccine administration, (2) current practices, and (3) factors associated with nonadherence. METHODS: An Internet-based survey was conducted in April 2008 among national samples. RESULTS: Response rates were 68% (220 of 325 physicians) among pediatricians and 51% (153 of 302 physicians) among family physicians. Seventy-three percent of pediatricians and 45% of family medicine physicians reported insufficient Hib vaccine supplies, and 22% to 24% reported having to defer doses for infants 2 to 6 months of age > or =10% of the time. Ninety-eight percent of pediatricians and 81% of family physicians were aware of the interim recommendations (P < or = .0001), and virtually all knew that the booster dose should be deferred; however, 22% of pediatricians and 33% of family medicine physicians reported not deferring this dose. Physicians in both specialties were less likely to adhere to recommendations to defer in this age group if they thought that their practice had sufficient vaccine supplies (pediatricians, odds ratio: 0.01 [95% confidence interval: 0.003-0.03]; family medicine physicians, odds ratio: 0.10 [95% confidence interval: 0.03-0.33]). Family medicine physicians were less likely to adhere to recommendations if they had not heard about the interim recommendations (odds ratio: 0.04 [95% confidence interval: 0.01-0.21]). CONCLUSIONS: Most primary care physicians experienced Hib vaccine shortages, and many have had to defer doses for 2- to 6-month-old children. Most are knowledgeable regarding interim recommendations, but one-fifth to one-third reported nonadherence.

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.

Suspected cervical spinal cord vascular anomaly in an African warthog (Phacochoerus africanus).

Vascular myelopathies of the spinal cord have not been described in Suidae, and are a rare finding in companion animals. An 8.5-yr female African warthog (Phacochoerus africanus) presented with an acute onset of tetraparesis. Based on neurologic findings, a cervical spinal cord lesion between C7-T2 was suspected. Magnetic resonance imaging revealed severe intramedullary hemorrhage with suspected abnormal vessels in the spinal cord at the level of the seventh cervical vertebrae. The acute onset of clinical signs and rapid deterioration of neurological status precluded surgical managements. A vascular anomaly was suspected on gross pathology and histology. Immunohistochemistry identified the lesion as a spontaneous intramedullary hematoma. Spontaneous intramedullary hematomyelia should be considered as a differential for acute onset of paresis in suid species.

Meloxicam and surgical denervation of the coxofemoral joint for the treatment of degenerative osteoarthritis in a Bengal tiger (Panthera tigris tigris).

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.