Hiatal hernia in a harbor seal (Phoca vitulina) pup

A 2-wk-old stranded harbor seal (Phoca vitulina) rescued by Mystic Aquarium showed signs of the presence of a hiatal hernia during rehabilitation. Contrast radiographs of esophagus and stomach revealed an intrathoracic radiodensity that contains filling defects typical of stomach, consistent with gastric rugal folds. Mural thickening was observed at the level of the cardia consistent with a diagnosis of a hiatal hernia. Although clinical improvement was noted with medical therapy and tube feeding, surgical correction of the hiatal hernia was considered necessary for full resolution. However, owing to the animal's low body weight, the corrective hernia surgery was postponed until the body condition improved. The seal needed to be surgically treated for a corneal ulcer, and while anesthetized with isoflurane, the seal became dyspneic and developed cardiac arrhythmias; ultimately cardiac arrest ensued. Resuscitation was unsuccessfully attempted and the seal was euthanized. Necropsy confirmed the radiographic diagnosis and further characterized a paraesophageal hiatal hernia.     

Analysis of blood gases,serum fat and serum protein: a new approach to estimate survival chances of stranded Harbor seal (Phoca vitulina) pups from the German North Sea

Background

Facing numerous challenges, such as illness, storms or human disturbance, some harbor seal (Phoca vitulina) pups lose contact to their dams and are found abandoned along the North Sea coast. In Schleswig-Holstein, pups with the prospect of surviving rehabilitation are admitted to the Seal Center Friedrichskoog. Despite elaborate clinical health assessments on admission, including differential hematology, in 2010, 17% of 108 admitted pups did not survive the first 20 days. The death rate during the years 2006 and 2009 varied between 9 and 19%. To broaden the spectrum of variables which could be predictive for survival, blood gas and serum analyses were performed for 99 pups using venous blood. Variables included total CO₂, pH, partial CO₂, HCO₃^–, base excess and anion gap as well as glucose, urea nitrogen, sodium, potassium and chloride. Moreover, total serum protein and fat (triglyceride) concentrations were measured for all pups on admission.

Results

Repeated measurements of 12 randomly selected individuals revealed a significant (p = 0.002) positive influence of time in rehabilitation on triglyceride concentrations. This trend probably shows the improvement of the pups’ nutritional status as a consequence of the shift from milk replacer formula to fish. No such positive influence was detected for total protein concentrations though. Hematologic values, including blood gases, were not predictive for survival.

Conclusions

For the first time blood gas values are reported in this study for a large sample size (N = 99) of seal pups (regardless of their health status). The ranges and medians calculated from the data can serve as a stepping stone towards the establishment of reference values for neonate harbor seals. However, future investigations on the development of blood gases in harbor seals with different health conditions and ages over time are necessary to allow for a better understanding of acid–base regulation in harbor seals.     

Seroprevalence of Toxoplasma gondii in North-eastern Atlantic harbor seal (Phoca vitulina vitulina) and grey seal (Halichoerus grypus)

Antibodies to Toxoplasma gondii were determined in serum samples from 47 grey seals (Halichoerus grypus) and 56 harbor seals (Phoca vitulina vitulina) from the Atlantic coasts of United Kingdom and France. Antibodies to T. gondii assayed by the modified agglutination test (MAT) were found in 14 (13.6%; IC(95%): 7.0-20.2) of 103 seals tested, with titres of 1:25 in 13 seals and 1:50 in 1 seal. Seroprevalence against T. gondii (MAT 1:25 or higher) was significantly higher in grey seals (23.4%) compared to harbor seals (5.4%). No significant differences were found between seroprevalence against T. gondii and sex, age or geographical locations. These results show natural exposure of European harbor and grey seals to T. gondii oocysts in the Atlantic Ocean. To the best of our knowledge, this is the first serological survey of T. gondii in European grey and harbor seals.     

Diagnosis and treatment of Sarcocystis neurona in a captive harbor seal (Phoca vitulina)

A captive harbor seal (Phoca vitulina) presented with partial anorexia, ataxia, and head bobbing, which progressed to complete anorexia, lethargy, and persistent whole-body intention tremors within several days. Response to treatment with ponazuril, serology, and cerebrospinal fluid analysis supported a diagnosis of Sarcocystis neurona. Analysis of serum levels for ponazuril indicated that therapeutic levels could be achieved at a dosage of 5 mg/kg p.o. s.i.d., whereas clinical response was improved at a dosage of 10 mg/kg. Several months after initiation of antiprotozoal therapy, the neurologic signs resolved, although rare intermittent tremors were seen with significant exertion.     

Prevalence of Coxiella burnetii and Brucella spp. in tissues from subsistence harvested northern fur seals (Callorhinus ursinus) of St. Paul Island,Alaska

Background

The northern fur seal (Callorhinus ursinus) is an important cultural and nutritional resource for the Aleut community on St. Paul Island Alaska. In recent years, an increasing number of zoonotic pathogens have been identified in the population, but the public health significance of these findings is unknown. To determine the prevalence of Coxiella burnetii and Brucella spp. in northern fur seal tissues, eight tissue types from 50 subsistence-harvested fur seals were tested for bacterial DNA by real-time polymerase chain reaction.

Findings

Of the 400 samples tested, only a single splenic sample was positive for Brucella spp. and the cycle threshold (ct) value was extremely high suggesting a low concentration of DNA within the tissue. C. burnetii DNA was not detected.

Conclusions

Findings suggest that the risk of humans contracting brucellosis or Q fever from the consumption of harvested northern fur seals is low.     

Comparison of the somatotropic axis in free-ranging and rehabilitated harbor seal pups (Phoca vitulina)

The somatotropic axis, including growth hormone (GH), insulin-like growth factor (IGF)-I, and IGF binding proteins (IGFBP), is a bridge between growth physiology, developmental age, and nutritional status in domestic animals. However, the importance of the somatotropic axis in nutrition, growth, and development of harbor seals has not been previously explored. Given the difficulty of conducting longitudinal studies in free-ranging harbor seals, this study focused on the potential use of harbor seals in rehabilitation facilities as a model for free-ranging seals. The purpose of this research was to compare concentrations of components of the somatotropic axis in free-ranging versus rehabilitated harbor seal pups. The hypothesis was that measurements of the somatotropic axis will be similar between individuals of comparable age and nutritional status (fasting versus feeding). To investigate this hypothesis, harbor seal pups (n=8) brought to The Marine Mammal Center (Sausalito, California, USA) or Mystic Aquarium (Mystic, Connecticut, U.S.A.) were initially assessed and determined to be healthy but abandoned. All pups were less than 2 wk of age upon arrival at rehabilitation facilities. Standard length was assessed at the time of arrival and again at release. Body mass was measured every week and blood samples were collected from each pup at 0, 4, and 8 wk of rehabilitation. Blood was collected and morphometrics assessed in free-ranging harbor seal pups (n=8) from the Gulf of Maine. Sera were analyzed for GH, IGF-I, and IGFBP concentrations. Concentrations of GH, IGF-I, and IGFBP-2 and -3 in rehabilitated pups were within a similar range compared with free-ranging pups when considered in the context of presumed nutrient intake. These data suggest that rehabilitated harbor seals may provide a useful model to investigate the effects of nutrient intake on growth and development of harbor seals, and will provide insight into phocid endocrinology and metabolism.     

Seroconversion and abortion in cattle experimentally infected with Brucella sp. isolated from a Pacific harbor seal (Phoca vitulina richardsi).

Previously unrecognized Brucella species have been isolated from a number of marine mammals, including harbor seals (Phoca vitulina richardsi) in the Puget Sound area of the state of Washington. Because of the presence of dairy herds in proximity to the harbor seal populations, a study was conducted to determine the effects of the harbor seal Brucella isolate in experimentally inoculated cattle. Six pregnant cattle were exposed by intravenous injection (n = 3) or intraconjunctival inoculation (n = 3). Two pregnant cows were intravenously injected with saline and served as controls. All of the cows receiving the Brucella seroconverted on 1 or more tests commonly used for the detection of Brucella abortus infection. Two of the cattle receiving the intravenous inoculation aborted, and brucellae were demonstrated in the fetuses and dams immediately following abortion. The remaining 4 Brucella-inoculated animals and their fetuses were culture negative for the organism at 14 weeks postinoculation. Results of this study indicate the marine mammal Brucella is capable of producing seroconversion and abortion in cattle but is less pathogenic in that species than B. abortus.     

Adaptations of subpalpebral lavage systems used for llamas (Lama glama) and a harbor seal (Phoca vitulina).

Subpalpebral lavage systems (SPLSs) were adapted for use in zoo llamas (Lama glama) and a wild harbor seal (Phoca vitulina) during therapy for severe ulcerative keratitis or corneal perforation. One llama presented with a melting corneal ulcer caused by Pseudomonas aeruginosa, which necessitated frequent application of a topical ophthalmic antibiotic. The lavage system was used routinely during the day and was connected to a balloon infusion system at night to allow for continuous medication administration. The ulcer healed soon after therapy was extended to include overnight treatment with the infusion system. A SPLS system was also combined with a balloon infusor during postoperative treatment of a second llama that had sustained a corneal perforation. Both llamas tolerated the infusor/lavage systems well and regained vision. One llama had minor conjunctival irritation from the SPLS that resolved quickly without treatment. Bilateral SPLS were placed in a wild harbor seal for treatment of severe ulcerative keratitis associated with Candida albicans infection. The seal tolerated the lavage systems well throughout 14 wk of their use in an aquatic environment with other seals. Partial detachment of the lavage systems from the skin of the seal occurred a few times during treatment and was easily corrected. Severe keratitis resolved with administration of antimicrobials through the lavage systems, and the seal was returned to the wild. The use of SPLSs alone or in ombination with balloon infusion systems warrants consideration for exotic, wild, and aquatic animals that cannot tolerate repetitive manual applications of topical ophthalmic medication.     

Fetus in fetu in a harbor seal (Phoca vitulina richardi): histopathologic, genetic, and toxicologic analysis

A young harbor seal (Phoca vitulina richardi), stranded on the coast of California, was found to have a 20-cm-diameter cranial cervical mass. Surgical excision revealed the subcutaneous mass to be covered in haired skin with multiple glabrous areas and structures resembling a jaw with tooth buds, eyelids, and a tail. The mass deformed the host pup's skull. Histologic examination revealed a complete vertebra in the tail, teeth in the jaw, and areas resembling tongue and larynx. Class 1 MHC sequences amplified by polymerase chain reaction from the mass and the host twin were identical. The mass was diagnosed as a fetus in fetu, a rare congenital anomaly in which 1 conjoined twin is completely enclosed in the body of the other twin. The host pup died, and no additional defects were found; however, blubber levels of persistent organic pollutants were high. The cause of the congenital anomaly in this pup is uncertain.     

Molecular cloning,purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

The Brucella mdh gene was successfully cloned and expressed in E. coli. The purified recombinant malate dehydrogenase protein (rMDH) was reactive to Brucella-positive bovine serum in the early stage, but not reactive in the middle or late stage, and was reactive to Brucella-positive mouse serum in the late stage, but not in the early or middle stage of infection. In addition, rMDH did not react with Brucella-negative bovine or mouse sera. These results suggest that rMDH has the potential for use as a specific antigen in serological diagnosis for early detection of bovine brucellosis.     

Gastrointestinal parasites in an isolated Norwegian population of wild red deer (Cervus elaphus)

Background

Thirteen red deer (Cervus elaphus), culled from the isolated population at the Mongstad Oil Refinery, Norway, were investigated for gastrointestinal helminths. These animals, enclosed by the refinery fence, do not have contact with other ruminants and have a high population density considering the available browsing area (1 km²) within the refinery site (3 km²). The population was estimated to be 110-130 at the time of culling.

Results

The helminth fauna among these sampled red deer was enumerated and species were identified based on morphology. Ostertagia leptospicularis/O. kolchida was detected in 83% [CI 55 - 95%], Spiculopteragia spiculoptera/S. mathevossiani in 92% [CI 65 - 99%] and Trichostrongylus axei in 42%, [CI 19 - 68%] of the abomasa examined. Characterisation of the intestinal parasite fauna revealed Capillaria bovis, Cooperia oncophora, Oesophagostomum venulosum, Trichuris globulosa and tapeworm fragments (presumed anoplocephalids) in seven individuals. Only one calf had an infection with more than one intestinal helminth (tapeworm fragment and Trichuris globulosa). The remaining six deer had single species intestinal infections.No significant age related trends were seen, with the exception of higher intensity of infection of T. axei in yearlings relative to other age classes. Assessment of abomasal parasite burden and body condition revealed no significant trends. In calves, statistically non-significant correlation was seen between increased parasite burden and decreased slaughter weight, whilst the opposite was seen in adults with the heaviest adults exhibiting the higher burdens. Given the small sample size the trends that were seen need further investigation. The parasite burden was aggregated with three adult red deer harbouring 75% of the total abomasal parasite count.

Conclusion

This isolated population was parasitised by a reduced subset of gastrointestinal nematodes typical of this cervid across an extensive geographic range in Eurasia. The intensity and abundance of abomasal nematodes was higher in this isolated population than reported in similar studies of red deer populations across Europe.     

Protective effects of recombinant Brucella abortus Omp28 against infection with a virulent strain of Brucella abortus 544 in mice

The outer membrane proteins (OMPs) of Brucella (B.) abortus have been extensively studied, but their immunogenicity and protective ability against B. abortus infection are still unclear. In the present study, B. abortus Omp28, a group 3 antigen, was amplified by PCR and cloned into a maltose fusion protein expression system. Recombinant Omp28 (rOmp28) was expressed in Escherichia coli and was then purified. Immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive mouse serum. Furthermore, humoral- or cell-mediated immune responses measured by the production of IgG1 or IgG2a in rOmp28-immunized mice and the ability of rOmp28 immunization to protect against B. abortus infection were evaluated in a mouse model. In the immunogenicity analysis, the mean titers of IgG1 and IgG2a produced by rOmp28-immunized mice were 20-fold higher than those of PBS-treated mice throughout the entire experimental period. Furthermore, spleen proliferation and bacterial burden in the spleen of rOmp28-immunized mice were approximately 1.5-fold lower than those of PBS-treated mice when challenged with virulent B. abortus. These findings suggest that rOmp28 from B. abortus is a good candidate for manufacturing an effective subunit vaccine against B. abortus infection in animals.     

Brucella pinnipedialis hooded seal (Cystophora cristata) strain in the mouse model with concurrent exposure to PCB 153

Brucellosis, a worldwide zoonosis, is linked to reproductive problems in primary hosts. A high proportion of Brucella-positive hooded seals (Cystophora cristata) have been detected in the declined Northeast Atlantic stock. High concentrations of polychlorinated biphenyls (PCBs) have also been discovered in top predators in the Arctic, including the hooded seal, PCB 153 being most abundant. The aim of this study was to assess the pathogenicity of Brucella pinnipedialis hooded seal strain in the mouse model and to evaluate the outcome of Brucella spp. infection after exposure of mice to PCB 153. BALB/c mice were infected with B. pinnipedialis hooded seal strain or Brucella suis 1330, and half from each group was exposed to PCB 153 through the diet. B. pinnipedialis showed a reduced pathogenicity in the mouse model as compared to B. suis 1330. Exposure to PCB 153 affected neither the immunological parameters, nor the outcome of the infection. Altogether this indicates that it is unlikely that B. pinnipedialis contribute to the decline of hooded seals in the Northeast Atlantic.     

Age estimation in the harbour seal (Phoca vitulina) based on the closure of skull sutures and synchondroses

Age estimation of individuals is an important tool for the management of marine mammals and is mostly done by microscopic analysis of growth layer groups (GLGs) in teeth (cementum and dentin) and bone. The present study evaluated the use of sutural and synchondrosal closure in the skull of the harbour seal (Phoca vitulina) as a potential non-destructive alternative for age estimation. For that, we scored the ectocranial closure of 12 selected sutures and synchondroses in a sample of 303 Eastern Atlantic harbour seals (P. v. vitulina; 138 males, 165 females), for which data on age at death were available, based on determined gross-morphological criteria in individuals younger than 1 year and cement layer analysis in canines in older individuals. A strong positive relationship between the sum of closure level scores (SCS) and age (males: r = 0.8797, females: r = 0.8825) was recorded, which was stronger than that for the relationship between age and condylobasal length (CBL, males: r = 0.7085, females: 0.7086) (all p-values <0.0001). In adult individuals (≥5 years), CBL was higher in males than females (p < 0.0001), while SCS did not significantly differ between the two sexes (p = 0.148). Our findings show that the analysis of sutural/synchondrosal closure is a valid alternative for age estimation in harbour seals, thereby confirming results of previous studies on other pinniped species.     

Detection of Brucella species in the milk of infected cattle,sheep, goats and camels by PCR

One hundred and three milk samples were collected from 52 cows, 21 ewes, 18 goats and 12 camels. The animals tested positive to at least one of the following: (1) standard tube agglutination test (SAT); (2) Rose Bengal plate test (RBPT); (3) milk ring test (MRT). All milk samples were examined by culture and single-step polymerase chain reaction (PCR) techniques for detection of Brucella species. The PCR assay amplified Brucella-DNA from 29 bovine milk samples, 10 from sheep, 13 from goats and one from a camel. The direct culture method detected Brucella organisms from 24 samples of cows' milk, 12 from sheep, 10 from goats and failed to detect any Brucella organisms from camels' milk. PCR detected up to 100 colony forming units (CFU) of B. abortus per millilitre of milk in 100% of diluted milk samples, and 1000 CFU of B. melitensis from 70% of milk samples. Although the overall sensitivity of the PCR was higher than the culture method, it should be possible to increase the sensitivity to detect lower numbers of Brucella organisms in field samples. The speed and sensitivity of the PCR assay suggest that this technique could be useful for detection of Brucella organisms in bovine milk, as well as in sheep, goat, and camels milk.     

Health status of a population of nutria (Myocastor coypus) living in a protected area in Italy

Ninety trapped nutria (Myocastor coypus) from a protected area of Piedmont (Italy), including the Po river, were examined for the prevalence for lesions in major viscera, selected serum antibodies and enteric bacteria. Coccidial lesions in the liver included cholangitis, calcification and necrosis. Renal lesions were nonsuppurative interstitial nephritis and a single case of renal adenocarcinoma. The lungs had a 41.1% prevalence of nonsuppurative interstitial pneumonia. Ten of 87 sera (11.5%) had antibodies against Leptospira bratislava, 3 of 87 (3.4%) against Leptospira ichterohaemorrhagiae, 15 of 41 (36.6%) against Toxoplasma gondii, and antibodies against encephalomyocarditis virus were detected in 5 of 78 sera (6.4%). All fecal samples were negative for Salmonella, Shigella, and Pseudomonas, and growth of enterobacteriaceae was in the normal range.     

A study on the effect of different rifle calibres in euthanisation of grey seals (Halichoerus grypus) in seal traps in the Baltic Sea

Background

In recent years, the euthanasia of seals has been discussed internationally and concern has been raised regarding the use of rifles, the effect of different calibres, and which calibres are sufficient for humane euthanasia. This study therefore investigated the effect of different firearm calibres on euthanasia of grey seals (Halichoerus grypus) in traps, and provides information for the development and refinement of regulations for hunting seals in the wild.

Findings

The effect of different calibres was studied in 19 seals shot in the head and neck at close range. All seals were necropsied and radiographed to characterize the injuries caused by the bullets. All tested calibres, 5.6 mm bullet diameter or larger, and .12 shotgun, were sufficiently effective to cause severe skull fractures, meningeal haemorrhages and instant death.

Conclusions

Rifles with 5.6 mm bullet diameter or larger, and a .12 shotgun loaded with a slug fired at close range to the head and neck of grey seals all caused instant death and can therefore be recommended for hunting seals in the wild.     

Management of a caseous lymphadenitis outbreak in a new Iberian ibex (Capra pyrenaica) stock reservoir

Background

In 2010, an Iberian ibex (Capra pyrenaica hispanica) stock reservoir was established for conservation purposes in north-eastern Spain. Eighteen ibexes were captured in the wild and housed in a 17 hectare enclosure. Once in captivity, a caseous lymphadenitis (CLA) outbreak occurred and ibex handlings were carried out at six-month intervals between 2010 and 2013 to perform health examinations and sampling. Treatment with a bacterin-based autovaccine and penicillin G benzatine was added during the third and subsequent handlings, when infection by Corynebacterium pseudotuberculosis was confirmed. Changes in lesion score, serum anti-C. pseudotuberculosis antibodies and haematological parameters were analyzed to assess captivity effects, disease emergence and treatment efficacy. Serum acute phase proteins (APP) Haptoglobin (Hp), Amyloid A (SAA) and Acid Soluble Glycoprotein (ASG) concentrations were also determined to evaluate their usefulness as indicators of clinical status.Once in captivity, 12 out of 14 ibexes (85.7%) seroconverted, preceding the emergence of clinical signs; moreover, TP, WBC, eosinophil and platelet cell counts increased while monocyte and basophil cell counts decreased. After treatment, casualties and fistulas disappeared and both packed cell volume (PCV) and haemoglobin concentration significantly increased. Hp, SAA and ASG values were under the limit of detection or showed no significant differences.

Conclusions

A role for captivity in contagion rate is suggested by the increase in antibody levels against C. pseudotuberculosis and the emergence of clinical signs. Although boosted by captivity, this is the first report of an outbreak of caseous lymphadenitis displaying high morbidity and mortality in wild ungulates. Treatment consisting of both vaccination and antibiotic therapy seemed to prevent mortality and alleviate disease severity, but was not reflected in the humoural response. Haematology and APP were not useful indicators in our study, perhaps due to the sampling frequency. Presumably endemic and irrelevant in the wild, this common disease of domestic small ruminants is complicating conservation efforts for the Iberian ibex in north-eastern Spain.