Otariid gammaherpesvirus 1 in South American fur seals (Arctocephalus australis) and a novel related herpesvirus in free-ranging South American sea lions (Otaria byronia): Prevalence and effects of age, sex, and sample type.

Otariid gammaherpesvirus 1 (OtGHV1) is associated with high rates of urogenital carcinoma in free-ranging California sea lions (Zalophus californianus; CSL), and until recently was reported only in the Northern Hemisphere. The objective of this study was to survey free-ranging South American sea lions (Otaria byronia; SASL) and South American fur seals (Arctocephalus australis: SAFS) in Punta San Juan, Peru for OtGHV1 and to determine prevalence characteristics. Twenty-one percent (14/67) of urogenital swabs collected over three years (2011, 2014, 2015) from live pinnipeds of both species tested positive with a pan-herpesvirus conventional PCR. Sequencing of SAFS amplicons revealed 100% homology to OtGHV1 at the DNA polymerase, glycoprotein B, and viral bcl2-like genes. Sequencing of SASL amplicons revealed a novel related virus, herein called Otariid gammaherpesvirus 8 (OtGHV8). For comparison of sample sites, urogenital, conjunctival, and oropharyngeal swabs collected from 136 live pinnipeds of both species at Punta San Juan between 2011-2018 were then assayed using quantitative PCR for a segment of the OtGHV1/8 DNA polymerase gene using a qPCR assay now determined to cross-react between the two viruses. In total, across both species, 38.6% (51/132) of urogenital swabs, 5.6% (4/71) of conjunctival swabs, and 1.1% (1/90) of oropharyngeal swabs were positive for OtGHV1/8, with SASL only positive on urogenital swabs. Results from SASL were complicated by the finding of OtGHV8, necessitating further study to determine prevalence of OtGHV1 versus OtGHV8 using an alternate assay. Results from SAFS suggest a potential relationship between OtGHV1 in SAFS and CSL. Though necropsy surveillance in SAFS is very limited, geographic patterns of OtGHV1-associated urogenital carcinoma in CSL and the tendency of herpesviruses to cause more detrimental disease in aberrant hosts suggests that it is possible that SAFS may be the definitive host of OtGHV1, which gives further insight into the diversity and phyogeography of this clade of related gammaherpesviruses.

Evaluation of Metal Partitioning across Humboldt Penguin (Spheniscus humboldti) Egg Components.

Humboldt Penguin (Spheniscus humboldti) population declines are attributable to several multifaceted anthropogenic impacts. At present, the exposure of Humboldt Penguins to high concentrations of heavy metals in the marine environment is a preeminent concern, due to mining along the Peruvian coast near key rookery sites. Metal and selenium concentrations were determined in eggs collected from September 2020 to April 2021 from a managed-care penguin population at the Brookfield Zoo to establish reference values for health indices conducted on wild populations. Concentrations of 16 elements, with emphasis on those found in mine efflux-arsenic, cadmium, copper, lead, mercury, selenium, and zinc-were assessed via inductively coupled plasma mass spectrometry in yolk, albumen, and eggshell. Data analyses indicate a clear delineation between egg constituents, with lipid-rich yolk displaying notably higher concentrations (µg/g) of arsenic (0.20 ± 0.064), chromium (0.086 ± 0.03), cobalt (0.01 ± 0.003), iron (238.65 ± 54.72), lead (0.32 ± 0.97), manganese (2.71 ± 0.66), molybdenum (0.57 ± 0.14), tin (3.29 ± 0.99), and zinc (64.03 ± 13.01) than other components (albumen and eggshell). These data confirm that heavy metals are partitioned differently across Humboldt Penguin egg components, which provides insight into the potential connection between embryonic nutrient source contamination and subsequent chick viability.

MEASUREMENT OF PERSISTENT ORGANIC POLLUTANTS, PERFLUORINATED COMPOUNDS, AND TOXIC METALS IN THE BLOOD OF HUMBOLDT PENGUINS (SPHENISCUS HUMBOLDTI) AT PUNTA SAN JUAN, PERU USING DRIED BLOOD SPOTS.

The Humboldt penguin (Spheniscus humboldti) population at the Punta San Juan Marine Protected Area in Peru is considered critical to the long-term sustainability of this endangered species in Peru. Exposure of the rookery to environmental toxicants is a mounting concern because of regional growth of industries and human populations. Whole blood samples were collected from 30 free-ranging penguins in 2011 as part of a broader population health monitoring program. Dried blood spots (DBS) containing 50 µl of blood were prepared and analyzed to assess exposure to five groups of environmental contaminants. Concentrations of elements arsenic, cadmium, iron, lead, mercury, selenium, and thallium were analyzed using inductively coupled plasma mass spectrometry. Persistent organic pollutant concentrations were measured using gas chromatography-tandem mass spectrometry to analyze organochlorine pesticides (OCP; p,p'-DDT, p,p'-DDE, ß-hexachlorocyclohexane, t-nonachlor, and oxychlordane), polychlorinated biphenyls (congeners 138 and 153), and polybrominated flame retardants (polybrominated biphenyl-153 and polybrominated diphenyl ether congeners 47 and 99). Per- and polyfluoroalkyl substances, including perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid were measured using liquid chromatography-tandem mass spectrometry. Results revealed low levels of exposure to these selected contaminants, at levels not considered to be of concern for wildlife health. DBS methodology was considered effective in a field-based setting for quantification of whole blood concentrations of environmental contaminants in penguins.

THE PHARMACOKINETICS AND PHARMACODYNAMICS OF ORAL PONAZURIL IN THE TREATMENT OF SYSTEMIC ISOSPOROSIS IN PASSERINE BIRDS.

Systemic isosporosis, previously atoxoplasmosis, is a significant cause of mortality in juvenile passerine birds. Recommended treatment regimens are empiric and vary in efficacy. The goal of this study was to determine the pharmacokinetics and pharmacodynamics of ponazuril for treatment of systemic isosporosis. Ponazuril, diluted with water to create an oral suspension (50 mg/ml), was administered (100 mg/kg) to 72 European starlings (Sturnus vulgaris) by a single dose via direct oral gavage (n = 24), a single dose injected into superworm larvae (Zophobas morio; n = 24), or a daily dose mixed with commercial dog food to top-dress feed for 5 d (n = 24). Peak plasma concentrations were 5.84, 2.46, and 9.13 µg/ml for the direct gavage, injected larvae, and top-dressed feed groups, respectively. With repeated dosing, mean plasma concentrations from the top-dressed feed group were maintained between 8.12 to 13.11 µg/ml. Results suggested ponazuril at a dosage of 100 mg/kg administered via direct gavage or top-dressed feed, but not via injected larvae, would exceed the concentrations needed to inhibit merogony of other apicomplexan parasites in cell culture (5 µg/ml). To assess the pharmacodynamics of this dose, seven passerine birds, red-vented bulbuls (Pycnonotus cafer; n = 2), blue-grey tanager (Thraupis episcopus; n = 1), and red-capped cardinals (Paroaria gularis; n = 4), were identified as shedders of systemic Isospora spp. via fecal qPCR. Birds were then treated with ponazuril (100 mg/kg) daily on top-dressed feed for 14 d. Fecal shedding was assessed via qPCR for 6 wk from the initiation of treatment. Treatment was associated with reduction in proportions of fecal shedding during the treatment period and the week following treatment, but shedding resumed in all birds by the end of sampling. Results support that treatment of breeding birds with 100 mg/kg ponazuril could reduce the shedding of active oocysts and decrease risk of clinical infection in susceptible juveniles.

Survival improvements of marine mammals in zoological institutions mirror historical advances in human longevity.

An intense public debate has fuelled governmental bans on marine mammals held in zoological institutions. The debate rests on the assumption that survival in zoological institutions has been and remains lower than in the wild, albeit the scientific evidence in support of this notion is equivocal. Here, we used statistical methods previously applied to assess historical improvements in human lifespan and data on 8864 individuals of four marine mammal species (harbour seal, Phoca vitulina; California sea lion, Zalophus californianus; polar bear, Ursus maritimus; common bottlenose dolphin, Tursiops truncatus) held in zoos from 1829 to 2020. We found that life expectancy increased up to 3.40 times, and first-year mortality declined up to 31%, during the last century in zoos. Moreover, the life expectancy of animals in zoos is currently 1.65-3.55 times longer than their wild counterparts. Like humans, these improvements have occurred concurrently with advances in management practices, crucial for population welfare. Science-based decisions will help effective legislative changes and ensure better implementation of animal care.

Agreement Study between Total Leukocyte Count Methodologies in South American Sea Lions (Otaria byronia) and Peruvian Fur Seals (Arctocephalus australis).

South American sea lions (Otaria byronia) and Peruvian fur seals (Arctocephalus australis) are sympatric species inhabiting the coastal Peruvian marine ecosystem. Declining abundance has prompted population health monitoring programs, including temporal monitoring of blood parameters. Several methods exist to determine total leukocyte count, yet no studies have evaluated agreement between methods in pinnipeds. We assessed agreement between total leukocyte counts determined by blood film estimate, Leuko-TIC, HemoCue, and UNOPETTE methods by using archival results from pinnipeds at Punta San Juan, Peru. Blood film estimates were prospectively performed, and resulting data were compared with retrospective leukocyte counts obtained from both species between 2009 and 2019 by using the other methodologies. Agreement in hematologic counts between methods was evaluated using Passing-Bablok regression and Bland-Altman plots (α=0.05). In total, 295 individuals (201 A. australis and 94 O. byronia) were included in the analysis. The blood film estimate method resulted in the highest leukocyte values (P<0.0001). Leuko-TIC counts were significantly higher than HemoCue counts (P<0.0001). Constant and proportional error was present in the agreement between the blood film estimate method and the other methods. Given the variation demonstrated between the different methodologies, additional research is needed to further evaluate agreement between these methodologies. The results underscore the importance of maintaining consistency in leukocyte count methodology for monitoring trends in population health over time. Method consistency may be the more important clinical consideration for assessing changes in leucocyte count over time and avoiding apparent changes depending on the methodology used.

DETECTION AND PREVALENCE OF SPHENISCID ALPHA-HERPESVIRUS-1 (SpAHV-1) IN A SAMPLE OF HUMBOLDT PENGUINS (SPHENISCUS HUMBOLDTI) AT PUNTA SAN JUAN, PERU.

Infections with herpesvirus have contributed to respiratory, enteric, and neurological disease reports in avian species worldwide. Herpesviruses have been detected in penguin species before but have not been studied extensively. To better understand the impact of these viruses in free-living populations, an initial retrospective survey was performed on a wild population of Humboldt penguins (Spheniscus humboldti) in the Punta San Juan Marine Protected Area, Peru (15°22'S, 75°12'W) using tracheal swabs collected from 28 penguins in 2016 and 34 penguins in 2018. DNA extracted from these swabs was analyzed using a consensus herpesviral PCR assay targeting the DNA polymerase gene, and positive samples were sequenced. A single sample from 2016 was positive for spheniscid alpha-herpesvirus-1 (SpAHV-1), establishing an overall sample prevalence of 1.6% (95% CI: 0-8.6%). The positive animal was an adult male that did not show any clinical signs of herpesviral infection and was otherwise healthy based on physical exam and laboratory findings. This is the first detection of a herpesvirus in penguins at Punta San Juan, Peru, and the first step toward characterizing the implications of SpAHV-1 for Humboldt penguins. This investigation highlights the importance of continual disease surveillance in wild populations over time to monitor for changes that may impact long-term population viability.

CLINICOPATHOLOGIC EVALUATION OF AN EX SITU POPULATION OF ADULT AFRICAN WHITE-BELLIED PANGOLIN (PHATAGINUS TRICUSPIS).

Death due to illegal trade and trafficking serves as the largest threat to pangolins in their geographic range countries. Appropriate veterinary care is essential to the welfare of sick or debilitated pangolins. Given the challenge of comprehensive physical examination of this species due to its unique anatomic features and poorly studied physiology, laboratory diagnostics can provide essential information to guide appropriate medical care. This retrospective study included previously analyzed blood and urine samples collected from adult (n = 17) African white-bellied pangolins (Phataginus tricuspis) under managed care that were sampled over a 4-yr period as part of routine veterinary examinations. Samples from clinically normal animals were used to summarize results for this species. Pangolins had a lymphocyte-predominant leukogram with a mean neutrophil to lymphocyte ratio of 0.83 (min-max: 0.13-4.81). Seasonal, sex, and age differences were present for several hematologic and serum biochemical analytes. Urine samples had a median urine specific gravity (USG) of 1.040 (min-max: 1.006-1.060), a neutral-to-basic urine pH (6.0-8.6), and contained mainly struvite crystals or infrequently calcium oxalate dihydrate crystals. Winter samples had higher USG than summer samples. The clinicopathologic data provided herein offers applicable information useful for the clinical management of pangolins.

Sympatric otariids increase trophic segregation in response to warming ocean conditions in Peruvian Humboldt Current System.

Determining trophic habits of predator communities is essential to measure interspecific interactions and response to environmental fluctuations. South American fur seals, Arctocephalus australis (SAFS) and sea lions Otaria byronia (SASL), coexist along the coasts of Peru. Recently, ocean warming events (2014-2017) that can decrease and impoverish prey biomass have occurred in the Peruvian Humboldt Current System. In this context, our aim was to assess the effect of warming events on long-term inter- and intra-specific niche segregation. We collected whisker from SAFS (55 females and 21 males) and SASL (14 females and 22 males) in Punta San Juan, Peru. We used δ13C and δ15N values serially archived in otariid whiskers to construct a monthly time series for 2005-2019. From the same period we used sea level anomaly records to determine shifts in the predominant oceanographic conditions using a change point analysis. Ellipse areas (SIBER) estimated niche width of species-sex groups and their overlap. We detected a shift in the environmental conditions marking two distinct periods (P1: January 2005-October 2013; P2: November 2013-December 2019). Reduction in δ15N in all groups during P2 suggests impoverished baseline values with bottom-up effects, a shift towards consuming lower trophic level prey, or both. Reduced overlap between all groups in P2 lends support of a more redundant assemblage during the colder P1 to a more trophically segregated assemblage during warmer P2. SASL females show the largest variation in response to the warming scenario (P2), reducing both ellipse area and δ15N mean values. Plasticity to adapt to changing environments and feeding on a more available food source without fishing pressure can be more advantageous for female SASL, albeit temporary trophic bottom-up effects. This helps explain larger population size of SASL in Peru, in contrast to the smaller and declining SAFS population.

Multi-species outbreak of SARS-CoV-2 Delta variant in a zoological institution, with the detection in two new families of carnivores.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has a worldwide distribution in humans and many other mammalian species. In late September 2021, 12 animals maintained by the Chicago Zoological Society's Brookfield Zoo were observed with variable clinical signs. The Delta variant of SARS-CoV-2 was detected in faeces and nasal swabs by qRT-PCR, including the first detection in animals from the families Procyonidae and Viverridae. Test positivity rate was 12.5% for 35 animals tested. All animals had been vaccinated with at least one dose of a recombinant vaccine designed for animals and all recovered with variable supportive treatment. Sequence analysis showed that six zoo animal strains were closely correlated with 18 human SARS-CoV-2 strains, suggestive of potential human-to-animal transmission events. This report documents the expanding host range of COVID-19 during the ongoing pandemic.

INVESTIGATION OF THE USE OF SERUM BIOMARKERS FOR THE DETECTION OF CARDIAC DISEASE IN MARINE MAMMALS.

Cardiac disease has been extensively documented in marine mammals; however, it remains difficult to diagnose antemortem. Assays measuring cardiac troponin I (cTnI) and N-terminal pro-brain natriuretic peptide (NT-proBNP) are used as sensitive and specific biomarkers of cardiac disease in many species, but have not been widely investigated in marine mammals. This study aimed to provide a set of reference values for cTnI and NT-proBNP in belugas (BW) (Delphinapterus leucas), sea otters (SO) (Enhydra lutris), Steller sea lions (SSL) (Eumetopias jubatus), and California sea lions (CSL) (Zalophus californianus) with and without cardiac disease, and to determine if these biomarkers are useful indicators of cardiac disease in these species. First, existing immunoassays for cTnI and NT-proBNP were successfully validated utilizing species-specific heart lysate spiked serum. Cohorts were defined by histopathology as animals with no evidence of cardiac disease ("control"), with confirmed cardiac disease ("disease"), and with concurrent renal and cardiac disease ("renal") for which serum samples were then analyzed. Serum concentration ranges for cTnI (ng/ml) and NT-proBNP (pmol/L) were determined for control and disease cohorts. There was significantly higher cTnI (P= 0.003) and NT-proBNP (P= 0.004) concentrations in the CSL disease cohort, as well as positive trends in BW, SO, and SSL disease cohorts that did not reach statistical significance. NT-proBNP concentrations were significantly higher in the CSL renal cohort compared with the control (P < 0.001) and disease (P= 0.007) cohorts. These results suggest that cTnI and NT-proBNP may be clinically useful in the antemortem diagnosis of cardiac disease in CSL, and warrant further investigation in BW, SO, and SL.

Complications associated with total hip arthroplasty in four large nondomestic felids.

CASE DESCRIPTION: A 9-year-old 37-kg sexually intact male snow leopard (Panthera uncia) with no history of lameness but radiographic evidence of right femoral subluxation and flattening of both femoral heads, 2 juvenile (< 1 year old) 25-kg sexually intact male cheetahs (Acinoynx jubatus) with unilateral hind limb lameness resulting from trauma, and an 11-year-old 110-kg sexually intact female Amur tiger (Panthera tigris altaica) with a 2-year history of left hip joint osteoarthritis were examined. CLINICAL FINDINGS: No clinically relevant clinical findings other than hip joint problems were identified. All 4 felids underwent staged bilateral (snow leopard) or unilateral (cheetahs and tiger [Panthera tigris]) total hip arthroplasty (THA). TREATMENT AND OUTCOME: In the snow leopard, both femoral THA components were found to be luxated 1 year after surgery. Treatment consisted of autogenous corticocancellous rib graft augmentation of the dorsal acetabular rims and synthetic suture capsulorrhaphies. The snow leopard lived for an additional 4 years with no additional THA-related complications. In the other 3 animals, catastrophic complications (luxation in the cheetahs and femoral fracture in the tiger) occurred shortly after THA. The THA implants were removed, and excision arthroplasty was performed. Long-term outcomes were good in all 3. CLINICAL RELEVANCE: Findings underscore the challenges associated with THA in large nondomestic felids. Given the high risk for early catastrophic failure as a result of luxation or fracture, plans must be made and resources must be available in case revision surgery or implant removal with excision arthroplasty becomes necessary.

Adenovirus Diversity in Fur Seal and Penguin Colonies of South America.

Adenoviruses are medium size nonenveloped viruses with a trend of coevolution with their hosts. We surveyed South American fur seals (Arctocephalus australis) and Humboldt penguins (Spheniscus humboldti) for adenoviruses at two sites from 2009 to 2012. Despite the common pattern of host specificity, some of the adenoviruses in our study were present in samples from unexpected host species. We identified mastadenoviruses, aviadenoviruses, and siadenoviruses in A. australis from Peru and Chile and in S. humboldti from Peru. The El Niño Southern Oscillation (ENSO) significantly reduces the productivity of the Humboldt upwelling system, which can change trophic and other ecological interactions, facilitating exposure to new pathogens. One aviadenovirus was detected in both the penguins and the fur seals, an interclass distance. This finding occurred only during the 2009 ENSO and not in 2010 or 2012. Further studies of viral diversity in sites with high-density mixed species populations are necessary to better understand viral evolution and the effect of environmental change on viral evolution and host specificity.

Detection of a novel herpesvirus associated with squamous cell carcinoma in a free-ranging Blanding's turtle.

The spread of both infectious and noninfectious diseases through wildlife populations is of increasing concern. Neoplastic diseases are rarely associated with population-level impacts in wildlife; however, impacts on individual health can be severe and might reflect deteriorating environmental conditions. An adult male free-ranging Blanding's turtle (Emydoidea blandingii) originally captured in 2005 and deemed healthy, was recaptured in 2018 with a 1 × 1.5 cm intra-oral broad-based right mandibular mass. An excisional biopsy was performed, and histopathology revealed squamous cell carcinoma (SCC). Consensus herpesvirus PCR identified a novel herpesvirus (proposed name Emydoidea herpesvirus 2 [EBHV-2]) within the tumor. EBHV-2 shares 85% sequence homology with Terrapene herpesvirus 2 (TerHV-2), a herpesvirus linked to fibropapillomas in eastern box turtles (Terrapene carolina carolina). Virus-associated fibropapillomas have been identified in multiple marine turtle species and have had debilitating effects on their populations, but to date, virus-associated SCCs are rarely reported.

MEASUREMENT OF INTRAOCULAR PRESSURE USING REBOUND TONOMETRY IN ANESTHETIZED PERUVIAN FUR SEALS (ARCTOCEPHALUS AUSTRALIS) FROM PUNTA SAN JUAN, PERU.

Ocular disease in pinnipeds under human care is well described, and intraocular pressure (IOP) can be impacted by a variety of ophthalmic conditions. Species-specific reference parameters from clinically normal animals are instrumental for understanding how ophthalmic diseases may impact ocular pressures. IOP measurements were obtained using rebound tonometry from free-ranging Peruvian fur seals (Arctocephalus australis unnamed subspecies) at Punta San Juan, Peru, over a 6-yr period (2010-2016). Retrospective data obtained from 108 (81 adults and 27 neonates comprising 69 females and 39 males) anesthetized fur seals with normal anterior segment ophthalmic examinations was included in the analysis. Differences in IOP from each eye were compared to categorical variables (age, year, sex, restraint) using an independent-samples t test. All univariate results with a significance of P < 0.05 were included in multivariate analysis. Of the 13 general linear models evaluated, the top two for both the right and the left eye included age class when all variables were evaluated simultaneously. Neonates had significantly lower IOP values than adults in both the right eye (17.5 mm Hg; 95% confidence interval [CI]: 14.0-21.1 mm Hg compared to 33.5 mm Hg; 95% CI: 31.0-36.1 mm Hg, respectively) and the left eye (18.4 mm Hg; 95% CI: 14.4-22.5 mm Hg compared to 32.3 mm Hg; 95% CI: 29.3-35.3 mm Hg, respectively). Anesthesia method was not statistically significant (P > 0.05). This is the first report of normal IOP measurements for any fur seal species. Described data can be used to improve diagnosis and management of ocular alterations in pinnipeds.

SUCCESSFUL SURGICAL CORRECTION OF PHIMOSIS IN AMUR LEOPARDS (PANTHERA PARDUS ORIENTALIS) USING A CARBON DIOXIDE LASER: CASE SERIES.

Three juvenile (<1 yr of age), genetically related Amur leopards (Panthera pardus orientalis) were diagnosed with phimosis. In all cases, no clinical signs were identified, and phimosis was detected during routine examination. Surgical enlargement of the preputial orifice was performed successfully using carbon dioxide laser with a 0.25-mm tip and an 8-watt continuous wave setting. A linear incision on the ventral aspect of the preputial opening allowed for complete preputial extrusion followed by placement of simple-interrupted sutures on both sides of the incision. No postoperative complications were observed, and there was no evidence of phimosis recurrence at subsequent examinations. Preputioplasty with carbon dioxide laser was used to successfully manage Amur leopards with phimosis. Given relatedness of these cases, a genetic predisposition for phimosis development in Amur leopards must be considered.

A QPCR ASSAY AND TESTING GUIDELINES FOR THE MOLECULAR DIAGNOSIS OF SYSTEMIC ISOSPOROSIS (FORMERLY ATOXPLASMOSIS) IN PASSERINE BIRDS.

Systemic isosporosis (formerly atoxoplasmosis), is a protozoal infection that causes death in nestling and fledgling passerine birds impacting ex situ breeding and reintroduction programs. Because current antemortem diagnostic tests lack sensitivity, a qPCR was developed for detection of Isospora spp. using primers and a fluorescent-tagged MGB probe targeting the large subunit (28s) ribosomal RNA gene (assay efficiency = >100%; sensitivity = <1 dsDNA copy). The assay was used to screen postmortem frozen or formalin-fixed paraffin-embedded tissue samples from passerine birds (n = 24; 12 with confirmed systemic isosporosis), whole blood and feces (n = 38) from live passerines, and other tissues infected with phylogenetically similar protozoa. The qPCR identified Isospora sp. DNA in tissues from 21/24 birds including 12/12 birds with cytologically-histologically confirmed infection (100% sensitivity) and 9/12 birds lacking microscopic organisms. The assay also amplified Eimeria sp. DNA; however, sequence analysis ruled out infection in the passerine cases. Blood and/or feces were positive in 30/38 birds, and in only 7/38 birds, blood and feces both contained Isospora sp. DNA. Finally, the qPCR was utilized to screen 30 consecutive daily fecal samples from live passerines (n = 20) to determine optimal sampling protocols. One or more of the daily fecal samples were positive in all 20 birds. In individual birds, the interval between positive qPCR amplification results ranged from 0 to 23 days, with an average of 5.85 days. Simulated application of 13 potential sample collection schedules was used to identify the sensitivity of repeated testing for identification of infected birds. Increased sampling days resulted in higher sensitivity but increased both cost and animal handling requirements. Based on statistical analysis and clinical considerations, the testing recommendation for detection of fecal shedding was collection and assay of five consecutive daily fecal samples, which had an average diagnostic sensitivity of 0.86.

Effects of nest type and sex on blood saccharide profiles in Humboldt penguins (Spheniscus humboldti): Implications for habitat conservation.

Reproductive success of endangered Humboldt penguin (Spheniscus humboldti) colonies in Peru has been associated with nesting habitat type, presumably due to differences in environmental exposure and activity patterns that may affect energy demands and metabolism. Gas chromatography and mass spectrometry were used to determine serum concentrations of 19 saccharides from 30 Humboldt penguins nesting at Punta San Juan, Peru in order to evaluate differences in metabolic state between penguins nesting in a sheltered burrow or crevice (n = 17) and those in exposed surface nests (n = 13). Univariate and multivariate statistical analyses identified serum saccharides (arabinose, maltose, glucose-6-phosphate, and levoglucosenone in particular) that were nest-dimorphic with substantial differences between surface- and sheltered-nesting penguins. Four sugars (arabinose, xylose, fructose-6-phosphate, and sucrose) had ≥ 2-fold difference in concentration between nest types. Seven saccharides were in the top five subsets generated by discriminant analysis; four of these are simple sugars (D-glucopyranose, α ⇄ ß; D-glucose; D-maltose; and D-mannose) and three are derivatives (glucose 6-phosphate, levoglucosenone, and N-acetylglucosamine). D-ribose had the highest information values (generated from weight-of-evidence values) followed by glucose 6-phosphate, levoglucosenone, and D-galactose. Sex was not a significant predictor of saccharide concentration. Levoglucosenone, which is a metabolite of the environmental contaminant levoglucosan, was significantly higher in surface-nesting penguins, reflecting a higher rate of exposure in non-sheltered penguins. Differences in the saccharide profiles of surface- and sheltered-nesting Humboldt penguins likely reflect increased metabolic requirements of surface-nesters at Punta San Juan. Conservation of appropriate sheltered-nesting habitat for penguins is essential for sustained reproductive success and colony health.

Circulating nutrient concentrations in free-ranging Humboldt penguins (Spheniscus humboldti) in Punta San Juan, Peru.

Free-ranging Humboldt penguin (HP, Spheniscus humboldti) populations are under pressure from resource competition with industrial fisheries, habitat loss, and El Niño Southern Oscillation events. Foraging patterns for this top marine predator change during periods of aberrant oceanographic conditions and scarce fish stock numbers. These radical dietary fluctuations can lead to poor fertility, early embryonic death, poor hatchability, suppressed immune function, high chick mortality, and illness. To understand the variability of nutrient status in reproductive seasons, we measured select circulating nutrient concentrations (fat-soluble vitamins A, D, E, and carotenoids, fatty acids, amino acids, minerals, and electrolytes) of 105 HP at Punta San Juan, Peru during the first reproductive seasons of 2007 and 2008. We determined significant differences in nutrient status between sexes, years of sampling, and reproductive stages. Males (4.5 ± 0.38 kg) weighed more than females (4.0 ± 0.29 kg) and exhibited higher concentrations of vitamin A (0.71 ± 0.11 vs. 0.61 ± 0.12 µg/ml) and docosahexaenoic acid (6.70 ± 1.61 vs. 5.65 ± 1.59%). Males also displayed lower concentrations of ß-carotene (0.01 ± 0.01 vs. 0.012 ± 0.001 µg/ml) and phosphorus (3.43 ± 0.83 vs. 4.40 ± 1.66 mg/dl). Comparison between the 2 years showed most circulating amino acid concentrations were higher in 2007. Significant differences in circulating amino acids and vitamins were also noted between different reproductive stages. These results demonstrate concentrations of nutrients can vary due to the physiological state of the animal, as well as the overall dynamics of their marine ecosystem habitat.

COMPUTED TOMOGRAPHY LUNG VOLUME DIFFERS BETWEEN VERTICAL AND INVERTED POSITIONING FOR EGYPTIAN FRUIT BATS (ROUSETTUS AEGYPTIACUS).

This prospective study characterizes the impact of positioning on the pulmonary volume and pulmonary atelectasis in Egyptian fruit bats (Rousettus aegyptiacus). The soft tissue appearance of atelectactic pulmonary parenchyma can obscure or mask pulmonary pathology. Soft tissue within healthy lung parenchyma caused by atelectasis can efface the margins of pathology, such as pulmonary metastasis or pneumonia, due to overlapping attenuation profiles. Pulmonary atelectasis is an unwanted side effect of anesthesia resulting from muscle relaxation and is exacerbated by high (80-100%) inspired oxygen supplementation during general anesthesia. Positioning can help minimize pulmonary atelectasis. Seven R. aegyptiacus received computed tomography imaging in suspended vertical (head-up) and inverted (head-down) positions that generated images in the dorsoventral plane. Vertically positioned bats had a significantly greater lung volume compared to inverted positioning (P = 0.0053). The nondependent portion of the lung apices in the vertically positioned bats had significantly more negative Hounsfield units (i.e. less dense tissue) than the dependent portions of the lung and was also less dense than both portions of the lungs in inverted positioned bats. Although not an intuitive positioning for bats, a vertical orientation generates less pulmonary atelectasis and a greater lung volume compared to bats positioned in a more natural inverted position. Despite physiologic adaptations to hang in an inverted position when not in flight, avoidance of inverted positioning during anesthesia and anesthetic recovery is recommended based on these findings.