Erysipelas vaccination protocols in dolphins Tursiops truncatus evaluated by antibody responses over twenty continuous years.

Erysipelas is an infection caused by Erysipelothrix rhusiopathiae that affects many different species around the world, including cetaceans. The acute septicemic form can rapidly cause death in bottlenose dolphins Tursiops truncatus. The ultimate goals of this long-term study were the development and identification of the most effective vaccination protocol against clinical erysipelas in T. truncatus using a commercially available swine vaccine, and to determine whether there is a need for a semi-annual vaccination versus an annual vaccination. The present study concentrated on the immunization of a dolphin population (7 wild-born and 22 captive-born individuals) with 2 swine vaccines, the European 'Eurovac Ery®' vaccine and the American 'ER Bac Plus®' vaccine, and immunological profile results over a 20-yr time period. The general protocol was a primo-vaccination (between 3 and 7 mo of age for calves) with or without a booster 1 mo post primo-vaccination and either annual or semi-annual vaccination thereafter. Sera were collected prior to vaccination, 2 wk post-vaccination and monthly. A dolphin-specific ELISA was developed to analyze the erysipelas-specific antibody response of vaccinated animals. The final ELISA results (n = 1362 samples from 29 animals at pre- and post-vaccination time) suggest that (1) there is a significant difference in antibody levels at the start of the vaccination between older and younger animals; (2) at least 3 vaccinations are necessary to obtain antibody levels above the levels at pre-vaccination; (3) thereafter, annual vaccinations seem sufficient to keep antibody levels above the levels at pre-vaccination; and (4) both vaccines induced similar responses. No case of erysipelas infection was observed in this population during the study.

SURGICAL TREATMENT OF OSTEOARTHRITIS IN HARBOR SEALS (PHOCA VITULINA).

In 2012, 543 harbor seals (Phoca vitulina) and 124 grey seals (Halichoerus grypus) were admitted to the Seal Rehabilitation and Research Centre in Pieterburen, The Netherlands. In 19 seals (3%), signs of infection in a hind flipper were observed. Initial treatment consisting of antibiotics and anti-inflammatory drugs resolved the symptoms in 15 animals. In four harbor seals, estimated to be 3 to 4 mo old, a necrotizing infection developed that resulted in osteoarthritis of the tarsus or tibiotarsal joint or both. Bacterial culture revealed the presence of polymicrobial infection in three of the four animals. Treatment consisted of amputation of the hind flipper under general anesthesia combined with tumescent anesthesia in the operation field. Amputations were done at the diaphysis of the tibia and fibula. After resecting these bones, the flipper was discarded, leaving a good muscle-skin cuff to cover the edges of the bones and close the skin without tension. The estimated blood loss varied between <50 to 150 ml. Healing was uneventful, and both antibiotics and analgesics were gradually reduced according to the individual response. The seals did not show any functional impairment 1 mo postoperatively. After release to the sea, scrutinous revision of all radiographs showed signs of osteomyelitis in at least one animal in the proximal part of the tibia, also present preoperatively. It is concluded that tumescent anesthesia in seals may reduce perioperative blood loss and that a lower leg amputation is a surgically easy and clean approach for the treatment of osteoarthritis of the hind flipper of seals, giving good functional results (diving, catching fish, exiting a pool, and moving on land).

Phylogenetic analysis of marine mammal herpesviruses.

Five novel DNA-dependent DNA polymerase (Dpol) herpesviral sequences were generated using nested consensus polymerase chain reaction (PCR) in clinical samples from a harbor seal (Phoca vitulina), bottlenose dolphin (Tursiops truncatus), orca (Orcinus orca), California sea lion (Zalophus californianus), and a Phocid herpesvirus 2 (PhHV-2) isolate from a harbor seal (used as positive control). These novel sequences and other representative herpesvirus sequences were included in Bayesian and Maximum Likelihood analyses to illustrate the phylogeny of herpesviruses amongst the marine mammal host species and in comparison to those of other animals. All 19 novel and known marine mammal herpesviruses included in the analyses aligned with members of the Alphaherpesvirinae or Gammaherpesvirinae subfamilies. The novel harbor seal herpesvirus clustered with members of the Macavirus genus, subfamily Gammaherpesvirinae. The novel bottlenose dolphin herpesvirus clustered together in a monophyletic group with another delphinid alphaherpesvirus but could not be associated with an established genus. The orca herpesvirus also clustered with a delphinid alphaherpesvirus and formed a separate clade. The sea lion herpesvirus clustered with PhHV-2. PhHV-1 clustered with varicelloviruses and PhHV-2 clustered strongly in the Gammaherpesvirinae genus Percavirus. All cetacean gammaherpesviruses formed a monophyletic clade and could not be associated with an established gammaherpesviral genus.

Ovarian interstitial cell tumor in a South American sea lion (Otaria flavescens).

A case of an ovarian tumor is reported in an 8-yr-old South American sea lion (Otaria flavescens) kept in a marine park in Malta (35.57 degrees N, 14.25 degrees E). The neoplasm was a solid mass of dense sheets and nests of round to polyhedral, irregularly shaped cells with abundant, finely vacuolated cytoplasm. The nuclei were uniformly small and round to oval. The supporting stroma contained thecal cells. The tumor cells were positive for positive inhibin and vimentin and focally positive for cytokeratin by immunohistochemistry. The neoplasm was diagnosed as an ovarian sex cord-stromal tumor, specifically an interstitial cell tumor.

Papillomavirus antibody prevalence in free-ranging and captive bottlenose dolphins (Tursiops truncatus).

Genital epithelial tumors of Atlantic bottlenose dolphins (Tursiops truncatus [Tt]) and Burmeister's porpoises (Phocoena spinipinnis) were formerly shown to be associated with papillomavirus (PV) infection. Papillomaviruses are highly prevalent viruses involved in the development of various tumor types in a wide range of animals, and so-called high-risk PVs contribute to malignant progression. In marine mammals, the incidence and prevalence of PV infection, transmission pathways, and persistence of infection are largely unknown. Using virus-like particles of bottlenose dolphin PV type 1 (TtPV1) as the antigen, enzyme-linked immunosorbent assay (ELISA) studies were conducted to evaluate PV antibody prevalence in bottlenose dolphins. In total, sera obtained from 115 dolphins were examined. Fifty-one percent of captive dolphins (n=18 of 35) and 90% of free-ranging dolphins (n=72 of 80) were antibody positive. Higher ELISA reactivity was observed among males compared with females. Sexually immature dolphins appeared more likely to seroconvert with age. Besides determining their PV antibody prevalence, each animal was also assessed for the presence of orogenital tumors. Interestingly, the mean age of free-ranging dolphins with tumors (n=21) was 11.2 yr compared with 29.9 yr in captive dolphins with tumors (n=9). Results from the current study suggest PV infection in bottlenose dolphins is common, that the main route of PV transmission among them may be horizontal, and that orogenital neoplasia may develop in early life stages of certain free-ranging bottlenose dolphins.

Genomic characterization of novel dolphin papillomaviruses provides indications for recombination within the Papillomaviridae.

Phylogenetic analysis of novel dolphin (Tursiops truncatus) papillomavirus sequences, TtPV1, -2, and -3, indicates that the early and late protein coding regions of their genomes differ in evolutionary history. Sliding window bootscan analysis showed a significant a change in phylogenetic clustering, in which the grouped sequences of TtPV1 and -3 move from a cluster with the Phocoena spinipinnis PsPV1 in the early region to a cluster with TtPV2 in the late region. This provides indications for a possible recombination event near the end of E2/beginning of L2. A second possible recombination site could be located near the end of L1, in the upstream regulatory region. Selection analysis by using maximum likelihood models of codon substitutions ruled out the possibility of intense selective pressure, acting asymmetrically on the viral genomes, as an alternative explanation for the observed difference in evolutionary history between the early and late genomic regions of these cetacean papillomaviruses.