Kristi L. West

Coinfection and Vertical Transmission of Brucella and Morbillivirus in a Neonatal Sperm Whale (Physeter macrocephalus) in Hawaii, USA

Abstract The viral genus Morbillivirus and the bacterial genus Brucella have emerged as important groups of pathogens that are known to affect cetacean health on a global scale, but neither pathogen has previously been reported from endangered sperm whales (Physeter macrocephalus). A female neonate sperm whale stranded alive and died near Laie on the island of Oahu, Hawaii, US, in May of 2011. Congestion of the cerebrum and enlarged lymph nodes were noted on the gross necropsy. Microscopic findings included lymphoid depletion, chronic meningitis, and pneumonia, suggesting an in utero infection. Cerebrum, lung, umbilicus, and select lymph nodes (tracheobronchial and mediastinal) were positive for Brucella by PCR. Brucella sp. was also cultured from the cerebrum and from mediastinal and tracheobronchial lymph nodes. Twelve different tissues were screened for Morbillivirus by reverse-transcriptase (RT)–PCR and select tissues by immunohistochemistry, but only the tracheobronchial lymph node and spleen were positive by RT-PCR. Pathologic findings observed were likely a result of Brucella, but Morbillivirus may have played a key role in immune suppression of the mother and calf. The in utero infection in this individual strongly supports vertical transmission of both pathogens.

Relaxin and progesterone during pregnancy and the post-partum period in association with live and stillborn calves in bottlenose dolphins (Tursiops truncatus)

The objectives of this study were to validate a relaxin and progesterone RIA for use in bottlenose dolphins, and quantify and characterize both hormones in extracts of placental tissue and serum collected during pregnancy and the post-partum period, and compare the results between dolphins with live and stillborn calves. In Experiment 1, validation of a heterologous relaxin and progesterone RIA involved specific displacement of antibody-bound radiolabeled human relaxin or progesterone in response to increasing volumes of pooled pregnant dolphin serum and amounts of respective hormone standards added to a fixed volume of serum. The displacement curves were considered parallel and additive relative to respective standard curves. In Experiment 2, immunoreactive relaxin and progesterone were detected in placental extracts and, in corresponding serum samples, concentrations of both hormones were higher during the pre-partum than post-partum periods. Circulatory concentrations of progesterone decreased (P < 0.05) from relatively high concentrations during early and mid-pregnancy to intermediate concentrations by late pregnancy (month effect, P < 0.0001) in dolphins with live births, whereas, in dolphins with stillbirths, the decrease in progesterone began earlier (month-by-birth status interaction, P < 0.007); mean concentrations were lower at mid- (37%, P < 0.06) and late (25%) pregnancy. Temporally, relaxin increased (P < 0.05) progressively from relatively low concentrations during early pregnancy to high concentrations during late pregnancy (month effect, P < 0.0001) and was not different between birth statuses (birth status effect, P = 0.76; month-by-birth status interaction, P = 0.17). Even though the interaction did not reach significance, mean relaxin concentrations were 42%, 29%, and 34% lower at early, mid-, and late pregnancy, respectively, in dolphins with stillbirths than in those with live births. In conclusion, the pregnancy-specific increase in serum concentrations of relaxin and lower concentrations of both relaxin and progesterone in association with stillbirths suggest the potential for relaxin to be used diagnostically to determine pregnancy status, and one or both hormones to be used to assess placental function, and, perhaps, fetal well-being in bottlenose dolphins and other cetaceans.

Steno bredanensis (Cetacea: Delphinidae)

Abstract Steno bredanensis (Cuvier in Lesson, 1828) is a small odontocete commonly called the rough-toothed dolphin. A slender, gray dolphin with a slightly darker cape, this species is most easily distinguished from other small delphinids by a gradually sloping forehead and a long rostrum. It is the only species in the genus Steno. Despite reports of sightings or stranded specimens from all tropical and subtropical oceans, the species is thought to typically occur in low abundance. The conservation status of S. bredanensis is poorly known.

Thyroid hormone concentrations in relation to age, sex, pregnancy, and perinatal loss in bottlenose dolphins (Tursiops truncatus).

This study evaluated circulating concentrations of thyroid hormones in relation to age, sex, pregnancy status, and perinatal loss in bottlenose dolphins (Tursiops truncatus) under human care. A total of 373 blood samples were collected from 60 individual dolphins housed at nine aquariums/oceanariums. Serum concentrations of total and free thyroxine (T4) and triiodothyronine (T3) were analyzed with commercial RIA kits validated for use with dolphins. While the effect of age was indicated by higher (P<0.0001) concentrations of total and free T4 and T3 in juveniles than adults, the effect of sex on thyroid hormones was inconclusive. The effect of pregnancy was indicated by higher (P<0.035) total and free T4 and T3 during early pregnancy compared to non-pregnancy. For both successful and unsuccessful pregnancy outcomes, maternal concentrations of thyroid hormones were highest during early, intermediate during mid, and lowest during late pregnancy (P<0.07 to P<0.0001). Compared to live and thriving births, concentrations of total and free T4 and total T3 were lower (P<0.08 to P<0.001) in dolphins with perinatal loss. Lower concentrations ranged from 10% to 14% during early, 11% to 18% during mid, and 23% to 37% during late pregnancy. In conclusion, the effects of age, reproductive status and stage of pregnancy on thyroid hormone concentrations are necessary factors to take into account when assessing thyroid gland function. Since perinatal loss may be associated with hypothyroidism in dolphins, analysis of serum T4 and T3 should be considered for those dolphins that have a history of pregnancy loss.

Investigation of an Immunoreactive Chorionic Gonadotropin-Like Substance in the Placenta, Serum, and Urine of Pregnant Bottlenose Dolphins (Tursiops truncatus)

This study was designed to test the hypothesis that an immunoreactive chorionic gonadotropin (CG)like substance is present in full-term dolphin placentas and to determine if CG immunoreactivity can be detected in corresponding serum and urine samples for potential application to diagnose pregnancy. Six placentas were collected immediately after parturition from four captive bottlenose dolphins in 2003, 2007 (Experiment 1), and 2011 (Experiment 2). Serum and urine were collected during early, middle, and late pregnancy from the same dolphins. In Experiment 1, an eCG radioimmunoassay (RIA) was used to analyze dilutions of supernatants from the homogenates of each placenta for eCG immunoreactivity, and a commercial hCG Enzyme-Linked Immunosorbent Assay (ELISA) was used to analyze individual serum samples and dilutions of pools of serum and urine for hCG immunoreactivity. Specific CG immunoreactivity was not detected above assay sensitivities in any of the supernatants of respective placental homogenates, including the highest concentrate (100 mg/mL), nor in any of the individual samples and pools of serum and urine. In Experiment 2, the highest placental homogenate was increased five-fold (500 mg/mL), sensitivity of the eCG RIA was increased six-fold, and a different combination of hCG antibodies was used in an alternative commercial “sandwich”-type ELISA. Despite the optimization, specific CG immunoreactivity in placental tissue and individual serum and urine samples was not detected above assay sensitivities. In conclusion, the hypothesis that an immunoreactive CG-like substance is present in full-term dolphin placentas was not supported. In addition, non-immunoreactivity of a CG-like substance in serum and urine samples collected during various stages of pregnancy precluded the development and application of a CG-based immunoassay for diagnosing pregnancy status in dolphins.

Using cytochrome P4501A1 expression in liver and blubber to understand effects of persistent organic pollutant exposure in stranded Pacific Island cetaceans

Elevated levels of persistent organic pollutants (POPs) have been reported in tropical Pacific Island cetaceans and their environment. In addition, recent health concerns in cetacean populations have warranted investigation into potential physiological effects from POP exposure for this region. Cytochrome P450 1A1 (CYP1A1) is a candidate for examining such effects. This well-studied biomarker of exposure and effect was examined in stranded cetacean liver using immunoblot (n = 39, 16 species) and blubber using immunohistochemistry (n = 23, 10 species). Paired tissue samples allowed for CYP1A1 comparisons not only between species but also within each individual animal to examine differences between tissue types. Liver CYP1A1 expression correlated positively and significantly with blubber concentrations of all POP categories (n = 39, p < 0.050) except octachlorostyrene and pentachlorobenzene (p > 0.100). Among Stenella species, liver CYP1A1 tissue expression was correlated negatively with the sum of all blubber layer endothelial cell CYP1A1 expression (n = 14, p = 0.049). Overall, elevated expression of liver CYP1A1 confirms its use as a biomarker of POP exposure to cetaceans stranded in the tropical Pacific basin.

BLOOD CHEMISTRY AND HEMATOLOGY VALUES IN HEALTHY AND REHABILITATED ROUGH-TOOTHED DOLPHINS (STENO BREDANENSIS)

ABSTRACT Rehabilitation efforts for live stranded marine mammals are guided by diagnostic measures of blood chemistry and hematology parameters obtained from each individual undergoing treatment. Despite the widespread use of blood parameters, reference values are not available in the literature from healthy rough-toothed dolphins (Steno bredanensis) with which to infer the health status of an animal. We examined serum or plasma chemistry and hematology data from 17 rough-toothed dolphins either housed at Dolphin Quest French Polynesia or during their rehabilitation at the Dolphin and Whale Hospital in Sarasota, Florida, US between 1994 and 2005. Blood parameters were compared among healthy animals, rehabilitation animals that were eventually released, and rehabilitation animals that died. This study indicated significant differences in many blood parameters for the poorly known rough-toothed dolphin that are likely to vary between healthy and sick animals. These included aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, bicarbonate, and globulins, which were greater in sick dolphins, and alkaline phosphatase and total protein which were greater in healthy individuals. Total white blood cell counts were lower in healthy animals as were the absolute numbers of neutrophils, monocytes, and eosinophils. Analysis of first blood sample levels for glucose, sodium, and erythrocyte sedimentation rate may have value for triage and prognostic evaluation.