EFFICACY OF HUMAN RECOMBINANT GRANULOCYTE COLONY-STIMULATING FACTOR (G-CSF, FILGRASTIM; NEUPOGEN®) IN NEUTROPENIC CETACEANS.

Neutrophils are one of the initial cell lines of protection against pathogens, and when their concentrations in the blood are low, animals are highly susceptible to infections. Neutropenia has been reported in cetaceans secondary to administration of systemic sulfa antibiotics or antifungal medications and severe, overwhelming infection. Filgrastim was administered to treat neutropenia over a 15-y period in 11 cetaceans comprising four species-beluga (Delphinapterus leucas, n = 1), bottlenose dolphin (Tursiops truncatus, n = 4), killer whale (Orcinus orca, n = 5), and short-finned pilot whale (Globicephala macrorhynchus, n = 1)] ranging in age from 1 wk to >24 y. Seven study animals received multiple doses (2-6). All animals responded to at least one dose (1-7 µg/kg) of parenteral filgrastim characterized by an increase in peripheral immature (band) neutrophils, segmented neutrophils, or both. In most cases (9/11), neutrophil counts increased within 48 h of a single dose. Duration of response varied but was at least 2 wk in eight of the 11 animals and 5-9 d in the remaining animals. No adverse reactions were observed in any cases.

Technique, postoperative complications, and visual outcomes of phacoemulsification cataract surgery in 21 penguins (27 eyes): 2011-2015.

OBJECTIVE: To describe surgical technique, postoperative complications, and visual outcome in penguins after phacoemulsification lens extraction surgery. ANIMALS STUDIED: Twenty-one penguins (27 eyes) that had phacoemulsification from 2011 to 2015 at Animal Eye Associates. Species included are as follows: 14 southern Rockhopper (18 eyes, 66.6%), 4 Gentoo (4 eyes, 19%), 2 King (3 eyes, 9.5%), and 1 Chinstrap penguin (2 eyes, 4.8%). Eleven of the penguins were females, and 10 were males with average age at the time of surgery being 27.5 years (range of 22-31 years). PROCEDURE: This is a retrospective study of phacoemulsification cataract surgery patients from 2011 to 2015. Visual outcome was evaluated by veterinary ophthalmologists at postoperative recheck examinations and subjectively by penguin keepers using individual bird surveys and paired t tests for statistical analysis. RESULTS: All eyes were functionally visual after surgery and at the time of last follow-up. Based on keeper surveys, 81% (17/21) of penguins showed immediate improvements in overall quality of life and 90% (19/21) of penguins exhibited improvement in mobility and behavior within their exhibit following cataract removal. Of the 14 penguins that received 1:5 intracameral atracurium during surgery, 10 (71.4%) had moderate mydriasis, 1 (7.1%) had minimal mydriasis, and 3 (21.4%) showed no effect to the pupil. Seventy percent of the cases had phacoemulsification times less than 60 seconds/eye; the mean time was 72 seconds. Sixteen eyes (59.3%) underwent anterior capsulotomy only, planned anterior and posterior capsulotomies were performed in 3 eyes (11.1%), and the entire lens capsule was removed due to capsular fibrosis and wrinkling in 8 eyes (29.6%). The most common short-term postoperative complication was temporary mild blepharospasm and/or epiphora, reported in 8 eyes (29.6%) from 7 penguins (33.3%). Long-term complications, 2-6 years postoperatively, included posterior synechiation resulting in dyscoria (10 of 24 eyes, 41.7%) and capsular fibrosis in all penguins with residual lens capsule (19/19, 100%). CONCLUSION: The prognosis for vision and behavioral improvement after phacoemulsification surgery in penguins is good, with low incidence of vision-threatening complications.

Evaluation of anti-Erysipelothrix rhusiopathiae IgG response in bottlenose dolphins Tursiops truncatus to a commercial pig vaccine.

Erysipelothrix rhusiopathiae is the causative agent of erysipeloid in humans and of erysipelas in various animals, including bottlenose dolphins Tursiops truncatus, in which an infection has the potential to cause peracute septicemia and death. The purpose of this study was to evaluate the efficacy of using an off-label porcine (ER BAC PLUS®, Zoetis) E. rhusiopathiae bactrin in a bottlenose dolphin vaccination program by determining the anti-E. rhusiopathiae antibody levels in vaccinated dolphins over a 10 yr period. Serum samples (n = 88) were analyzed using a modified fluorescent microbead immunoassay from 54 dolphins, including 3 individuals with no history of vaccination and 51 dolphins with an average of 5 vaccinations, 3 of which had previously recovered from a natural E. rhusiopathiae infection. A mean 311-fold increase in the immunoglobulin G (IgG) antibody index was measured in a subsample of 10 dolphins 14 d after the first booster vaccination. Serum IgG antibody titers were influenced by number of vaccines received (r2 = 0.47, p < 0.05) but not by age, gender, history of natural infection, adverse vaccine reaction, vaccination interval or time since last vaccination. The commercial pig bacterin was deemed effective in generating humoral immunity against E. rhusiopathiae in dolphins. However, since the probability of an adverse reaction toward the vaccine was moderately correlated (p = 0.07, r2 = 0.1) with number of vaccines administered, more research is needed to determine the optimal vaccination interval.

DNA methylation networks underlying mammalian traits.

Using DNA methylation profiles (n = 15,456) from 348 mammalian species, we constructed phyloepigenetic trees that bear marked similarities to traditional phylogenetic ones. Using unsupervised clustering across all samples, we identified 55 distinct cytosine modules, of which 30 are related to traits such as maximum life span, adult weight, age, sex, and human mortality risk. Maximum life span is associated with methylation levels in HOXL subclass homeobox genes and developmental processes and is potentially regulated by pluripotency transcription factors. The methylation state of some modules responds to perturbations such as caloric restriction, ablation of growth hormone receptors, consumption of high-fat diets, and expression of Yamanaka factors. This study reveals an intertwined evolution of the genome and epigenome that mediates the biological characteristics and traits of different mammalian species.

Growth, maturity, reproduction, and life expectancy in ex-situ pacific walruses (Odobenus rosmarus divergens).

BACKGROUND: Pacific walruses are found in Arctic regions of the Chukchi and Bering Sea where rapid changes in environmental conditions resulting in loss of sea ice are occurring. Therefore, accurate life history data are crucial for species management plans and longitudinal data collected over the lives of individual walruses housed in zoos and aquaria provide otherwise difficult to obtain biological information. RESULTS: While similar at birth, Gompertz regression curves indicated that males grew faster than females (p < 0.0001) in weight (99 kg vs 57.6 kg/y) and length (26.9 cm vs 26.3 cm/y) with physical differences being detected by age 3 for weight and age 7 for length. Males reached adult weight at 13.5 ± 3.3 y and females by age 12.3 ± 2.3 y. The mean age at first ovulation and at first conception occurred at 8.8 y and 9.6 y. Greater than 75% of all conceptions and calving occurred between February and March and from May to June, respectively. Mean gestation lasted 423 d and false pregnancies lasted at least 169 d with a decrease (p < 0.05) in serum progesterone concentration between false pregnancy and pregnancy occurring within 6 months after ovulation. Based on these results, we estimated embryonic diapause to last from 120 to 139 days, and fetal growth last ~ 284 days. All males older than 8 y had an increase in serum testosterone and body weight that was highest in February and lowest in July. Overall, no differences were observed between male and female survival, with a mean (± SEM) life expectancy of 19.5 ± 1.5 y, respectively. Currently, the oldest male and female captive walruses are 40 and 43 y, respectively. CONCLUSIONS: Data provided herein include details of life history characteristics of zoo and aquaria housed walruses that are useful for wild population recovery models. In particular, results on survivorship and the identification of the most vulnerable period for calf survival can help with model development and suggests that for recovery to occur birthing locations for this species must be protected.

Multi-species and multi-tissue methylation clocks for age estimation in toothed whales and dolphins.

The development of a precise blood or skin tissue DNA Epigenetic Aging Clock for Odontocete (OEAC) would solve current age estimation inaccuracies for wild odontocetes. Therefore, we determined genome-wide DNA methylation profiles using a custom array (HorvathMammalMethyl40) across skin and blood samples (n = 446) from known age animals representing nine odontocete species within 4 phylogenetic families to identify age associated CG dinucleotides (CpGs). The top CpGs were used to create a cross-validated OEAC clock which was highly correlated for individuals (r = 0.94) and for unique species (median r = 0.93). Finally, we applied the OEAC for estimating the age and sex of 22 wild Norwegian killer whales. DNA methylation patterns of age associated CpGs are highly conserved across odontocetes. These similarities allowed us to develop an odontocete epigenetic aging clock (OEAC) which can be used for species conservation efforts by provide a mechanism for estimating the age of free ranging odontocetes from either blood or skin samples.

Ultrasonographic and hormonal characterization of reproductive health and disease in wild, semiwild, and aquarium-housed southern stingrays (Hypanus americanus).

OBJECTIVE: To characterize physical examination, plasma biochemical, and ultrasonographic findings in aquarium-housed, managed semiwild, and wild southern stingrays (Hypanus americanus) with and without reproductive disease. ANIMALS: Southern stingrays from aquarium (n = 48), lagoon (managed semiwild; 34), and wild (12) habitats. PROCEDURES: Limited, opportunistic prosections were performed of presumed anatomically normal wild southern stingrays and compared with findings for aquarium-housed stingrays with reproductive disease. Ultrasonographic video data from both groups were used to assign a score (1 to 5) indicating increasing severity of ovarian and uterine reproductive disease. Plasma total 17ß-estradiol, estrone, progesterone, and testosterone concentrations were measured with enzyme immunoassays validated for use in southern stingrays. RESULTS: Ultrasonographic ovarian scores were significantly correlated with uterine scores. No reproductive disease was detected in semiwild or wild stingrays, but 65% (31/48) of aquarium-housed stingrays had developing or advanced reproductive disease (ie, ultrasonographic ovarian or uterine score of 4 or 5). Significant correlations were identified between ovarian and uterine disease status and plasma concentrations of all steroid hormones except testosterone. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggested that ultrasonography and plasma hormone concentrations may be useful in the identification of reproductive disease and determination of disease severity in southern stingrays.