Advances in research on the impacts of anti-submarine sonar on beaked whales.

Mass stranding events (MSEs) of beaked whales (BWs) were extremely rare prior to the 1960s but increased markedly after the development of naval mid-frequency active sonar (MFAS). The temporal and spatial associations between atypical BW MSEs and naval exercises were first observed in the Canary Islands, Spain, in the mid-1980s. Further research on BWs stranded in association with naval exercises demonstrated pathological findings consistent with decompression sickness (DCS). A 2004 ban on MFASs around the Canary Islands successfully prevented additional BW MSEs in the region, but atypical MSEs have continued in other places of the world, especially in the Mediterranean Sea, with examined individuals showing DCS. A workshop held in Fuerteventura, Canary Islands, in September 2017 reviewed current knowledge on BW atypical MSEs associated with MFAS. Our review suggests that the effects of MFAS on BWs vary among individuals or populations, and predisposing factors may contribute to individual outcomes. Spatial management specific to BW habitat, such as the MFAS ban in the Canary Islands, has proven to be an effective mitigation tool and mitigation measures should be established in other areas taking into consideration known population-level information.

An epidemiological investigation of an idiopathic myopathy in hunting dogs in New Zealand.

AIM: To conduct an epidemiological investigation of an idiopathic myopathy, known as "Go Slow" (GSM), which was initially recognised in dogs used for pig hunting. A secondary aim was to describe the hunting activities, diet and health of dogs used for pig hunting in New Zealand. METHODS: A retrospective cohort study was conducted between June 2014-June 2017. Cases of GSM in dogs were diagnosed by veterinarians using a combination of clinical history, physical examination findings, serum biochemistry and/or skeletal muscle histology. A telephone interview was conducted with the owner or primary veterinarian to provide information regarding the dog's diet and exercise over the 7 days preceding the onset of clinical signs. In August 2015, a separate online survey of owners of dogs used for pig hunting was conducted to characterise the normal hunting activities, diet and health of these dogs. RESULTS: A total of 86 cases of GSM were recruited, of which 58 (67%) were pig hunting dogs, 16 (19%) pet dogs and 12 (14%) working farm dogs. Cases were most commonly reported in the upper North Island, and 65 (76 (95% CI=67-85)%) were from the Northland region. Processed commercial dog food had been fed to 93 (95% CI=88-98)% of affected dogs. Ingestion of raw, frozen or cooked wild pig in the preceding week was reported for 76 (88 (95% CI=82-95)%) dogs with the myopathy. In the survey of owners of healthy pig hunting dogs, 203 eligible responses were received; pig hunting was reported to most commonly occur in Northland (20.2%), Waikato (22.3%) and Bay of Plenty (23.2%) regions. Commercial dog food was fed to 172 (85 (95% CI=80-90)%) of the dogs included in this survey, and 55 (27 (95% CI=20-33)%) had eaten wild pig in the preceding week. The most common reported health problem in pig hunting dogs was traumatic wounds. CONCLUSIONS: Cases of GSM were most commonly recognised in dogs used for pig hunting, but also occurred in pet and working farm dogs. The disease was most frequently reported in the upper North Island of New Zealand and ingestion of wild pig was a consistent feature in cases of this myopathy. CLINICAL RELEVANCE: To minimise the risk of dogs developing this myopathy, it would seem prudent to avoid feeding any tissues from wild pigs to dogs in areas where the disease is known to occur.

First report of systemic toxoplasmosis in a New Zealand sea lion (Phocarctos hookeri).

CASE HISTORY: A 1-year-old female New Zealand sea lion (Phocarctos hookeri) was intermittently observed in the Otago region of New Zealand over an 11-month period, always dragging her hind flippers. In December 2012 the sea lion was found dead, after a period of several days being observed to be harassed by male sea lions. PATHOLOGICAL FINDINGS: At gross postmortem examination the sea lion was in moderate body condition with signs of recent bite wounds and bruising. The lungs were dark and poorly inflated. Histological findings included meningoencephalomyelitis, radiculomyelitis of the cauda equina, myocarditis and myositis. Toxoplasmosis gondii organisms were detected histologically and following immunohistochemistry in the brain, spinal cord, spinal nerves and pelvic muscles. MOLECULAR BIOLOGY: Nested PCR analysis and sequencing confirmed the presence of T. gondii DNA in uterine and lung tissue. A variant type II T. gondii genotype was identified using multilocus PCR-restriction fragment length polymorphism analysis. DIAGNOSIS: Systemic toxoplasmosis. CLINICAL RELEVANCE: Infection with T. gondii involving the spinal cord and nerves was the likely cause of the paresis observed in this sea lion before death. Ultimately, death was attributed to crushing and asphyxiation by a male sea lion, presumably predisposed by impaired mobility. Diagnosis of toxoplasmosis in a New Zealand sea lion highlights the possibility that this disease could play a role in morbidity and mortality in this endangered species, particularly in the recently established mainland populations that are close to feline sources of T. gondii oocysts.

Seroprevalence of Toxoplasma gondii in mainland and sub-Antarctic New Zealand sea lion (Phocarctos hookeri) populations.

AIMS: To investigate the seroprevalence of antibodies to Toxoplasma gondii in New Zealand sea lions (Phocarctos hookeri), as a potential contributor to reproductive failure. METHODS: Archived sera were sourced from New Zealand sea lions from two recolonising mainland populations in the Otago Peninsula (n=15) and Stewart Island (n=12), as well as a declining population at Enderby Island (n=28) in the New Zealand sub-Antarctic. Sera were tested for antibodies to T. gondii using a commercially available ELISA (with samples considered positive if the sample to positive ratio was >30%), and latex agglutination test (LAT; with titres ≥1:32 considered positive). Western blot analysis was used to validate the results of a subset of 14 samples. RESULTS: Five samples from sea lions in mainland locations were confirmed positive for antibodies to T. gondii. Two adult females exhibited high LAT antibody titres (min 1:2048, max 1:4096) on both occasions when sampled 1 and 2 years apart, respectively. No animals from Enderby Island were seropositive. CONCLUSIONS: Toxoplasma gondii infection is unlikely to be a major contributor to poor reproductive success in New Zealand sea lions. However, continued surveillance is pertinent to assess subclinical and clinical impacts of the parasite on these threatened populations. The commercial tests evaluated here, with further species-specific threshold refinement could provide a fast, inexpensive and reliable indicator of T. gondii exposure in New Zealand sea lions.

Brucellosis associated with stillbirth in a bottlenose dolphin in Australia.

A captive adult female bottlenose dolphin presented with stillbirth. The placenta appeared oedematous. No other gross lesions were evident in the placenta or the stillborn calf. Histopathology revealed mild multifocal placentitis and foetal encephalitis. Brucella sp. was isolated from lung, liver, spleen and kidney. Sequence and phylogenetic analysis demonstrated this organism to be most similar to Brucella ceti sequence type (ST) 27. Brucella sp. DNA was detected in formalin-fixed paraffin-embedded placenta and brain by real-time PCR using primers targeting the IS711 gene. Immunohistochemical staining revealed Brucella sp. antigen in placental inflammation. This is the first report of isolation of Brucella sp. from a marine mammal in the Southern Hemisphere and the first report of marine Brucella-associated disease in Australia.

Untargeted metabolic profiling of dogs with a suspected toxic mitochondrial myopathy using liquid chromatography-mass spectrometry.

'Go Slow myopathy' (GSM) is a suspected toxic myopathy in dogs that primarily occurs in the North Island of New Zealand, and affected dogs usually have a history of consuming meat, offal or bones from wild pigs (including previously frozen and/or cooked meat). Previous epidemiological and pathological studies on GSM have demonstrated that changes in mitochondrial structure and function are most likely caused by an environmental toxin that dogs are exposed to through the ingestion of wild pig. The disease has clinical, histological and biochemical similarities to poisoning in people and animals from the plant Ageratina altissima (white snakeroot). Aqueous and lipid extracts were prepared from liver samples of 24 clinically normal dogs and 15 dogs with GSM for untargeted liquid chromatography-mass spectrometry. Group-wise comparisons of mass spectral data revealed 38 features that were significantly different (FDR<0.05) between normal dogs and those with GSM in aqueous extracts, and 316 significantly different features in lipid extracts. No definitive cause of the myopathy was identified, but alkaloids derived from several plant species were among the possible identities of features that were more abundant in liver samples from affected dogs compared to normal dogs. Mass spectral data also revealed that dogs with GSM have reduced hepatic phospholipid and sphingolipid concentrations relative to normal dogs. In addition, affected dogs had changes in the abundance of kynurenic acid, various dicarboxylic acids and N-acetylated branch chain amino acids, suggestive of mitochondrial dysfunction.

Causes of adult mortality in two populations of New Zealand sea lions (Phocarctos hookeri).

The New Zealand sea lion is an endangered species endemic to New Zealand. While causes of death are well described for pups of this species, mortality in adults is poorly characterised. This study investigated causes of death in 136 New Zealand sea lions in two different populations: a major breeding site on remote, uninhabited Enderby Island in the sub-Antarctic, and a slowly increasing recolonising population on the inhabited mainland. For animals with at least a partial diagnostic investigation (n = 112), the most frequently diagnosed causes of mortality were infectious disease (41/112; 37%), particularly tuberculosis due to M. pinnipedii (20/112; 18%), and conspecific trauma (27/112; 24%). Anthropogenic trauma was an important cause of death in mainland sea lions (9/33; 26%). Deliberate anthropogenic mortality has previously been identified as the greatest potential threat to population recovery for mainland sea lions, and as human and pinniped populations increase, managing interactions between these species will become increasingly important.

Septicaemia and meningitis caused by infection of New Zealand sea lion pups with a hypermucoviscous strain of Klebsiella pneumoniae.

This study describes a syndrome of neonatal septicemia and meningitis in New Zealand sea lions, caused by a strain of Klebsiella pneumoniae that is phenotypically similar to strains causing environmentally-acquired septicemia and neuro-invasive disease in humans. Between late 2006 and early 2010, 123 pups from the Enderby Island breeding colony died of K. pneumoniae infection, with lesions including fibrinous to fibrinosuppurative meningitis, subdural hemorrhage, septic arthritis, herniation and hemorrhage of the cerebellar vermis, lymphadenitis and cellulitis. This infection was responsible for 58% of observed pup mortality over this time period, with most deaths occurring in the latter part of the breeding season (mid February onwards). The results of this study suggest that the pattern of this disease has changed since it was first described in 2002, when most deaths occurred early in the season (early to mid-January), and that it is an important and consistent cause of pup mortality in this population. In addition, a similar disease syndrome and bacterial strain was diagnosed in a single pup in a fragile recolonizing New Zealand sea lion population on mainland New Zealand, and the potential effect on this population is unknown but could have a negative impact on recolonisation at this site.

Traumatic brain injury, axonal injury and shaking in New Zealand sea lion pups.

Trauma is a common cause of death in neonatal New Zealand sea lion pups, and subadult male sea lions have been observed picking up and violently shaking some pups. In humans, axonal injury is a common result of traumatic brain injury, and can be due to direct trauma to axons or to ischaemic damage secondary to trauma. 'Shaken baby syndrome', which has been described in human infants, is characterised by retinal and intracranial subdural haemorrhages, and has been associated with axonal injury to the brain, spinal cord and optic nerve. This study identifies mechanisms of traumatic brain injury in New Zealand sea lion pups, including impact injuries and shaking-type injuries, and identifies gross lesions of head trauma in 22/36 sea lion pups found dead at a breeding site in the Auckland Islands. Despite the high frequency of such gross lesions, only three of the pups had died of traumatic brain injury. Observational studies confirmed that shaking of pups occurred, but none were shown to die as a direct result of these shaking events. Axonal injury was evaluated in all 36 pup brains using ß-amyloid precursor protein immunohistochemistry. Immunoreactive axons were present in the brains of all pups examined including seven with vascular axonal injury and two with diffuse axonal injury, but the severity and pattern of injury was not reliably associated with death due to traumatic brain injury. No dead pups had the typical combination of gross lesions and immunohistochemical findings that would conform to descriptions of 'shaken baby syndrome'. Axonal injury was present in the optic nerves of most pups, irrespective of cause of death, but was associated with ischaemia rather than trauma.

An atypical genotype of Toxoplasma gondii as a cause of mortality in Hector's dolphins (Cephalorhynchus hectori).

Hector's dolphins (Cephalorhynchus hectori) are a small endangered coastal species that are endemic to New Zealand. Anthropogenic factors, particularly accidental capture in fishing nets, are believed to be the biggest threat to survival of this species. The role of infectious disease as a cause of mortality has not previously been well investigated. This study investigates Toxoplasma gondii infection in Hector's dolphins, finding that 7 of 28 (25%) dolphins examined died due to disseminated toxoplasmosis, including 2 of 3 Maui's dolphins, a critically endangered sub-species. A further 10 dolphins had one or more tissues that were positive for the presence of T. gondii DNA using PCR. Genotyping revealed that 7 of 8 successfully amplified isolates were an atypical Type II genotype. Fatal cases had necrotising and haemorrhagic lesions in the lung (n=7), lymph nodes (n=6), liver (n=4) and adrenals (n=3). Tachyzoites and tissue cysts were present in other organs including the brain (n=5), heart (n=1), stomach (n=1) and uterus (n=1) with minimal associated inflammatory response. One dolphin had a marked suppurative metritis in the presence of numerous intra-epithelial tachyzoites. No dolphins had underlying morbillivirus infection. This study provides the first evidence that infectious agents could be important in the population decline of this species, and highlights the need for further research into the route of entry of T. gondii organisms into the marine environment worldwide.

Freezing and thawing of pinniped carcasses results in artefacts that resemble traumatic lesions.

The objective of this study was to assess whether the freezing and thawing of pinniped carcasses prior to post-mortem examination could create artefacts that resembled lesions caused by trauma. Necropsy findings in New Zealand fur seals (Arctocephalus forsteri), captured incidental to commercial fishing, and either chilled (n=5) or frozen (n=5), were compared. Changes in frozen, but not in chilled, carcasses included: pseudo-bruising of subcutis; the accumulation of thick dark red fluid (resembling haemorrhage) in the thoracic cavity, pericardial sac and abdominal cavity; apparent subcapsular renal haemorrhage; pseudo-contusions of the brain; apparent haemorrhage from the nares; and blood-staining of the anterior ocular chamber. The processes of freezing and thawing were strongly associated with subcutaneous pseudo-bruises, the presence of thick, dark red abdominal fluid and renal subcapsular 'haemorrhage' (P=0.004). These artefacts probably develop due to a combination of autolysis and 'freeze-thaw' effects including lysis of cell membranes, fluid shifts into the extracellular space, and disruption of blood vessel walls. The results of the study demonstrate that artefacts resembling traumatic lesions are created during freezing and thawing of pinniped bodies. Such changes must be taken into consideration at post-mortem examination of previously frozen carcasses.

Bilateral ovarian adenocarcinoma in a mare causing haemoperitoneum and colic.

CASE HISTORY: A 13-year-old Thoroughbred mare was presented with a history of mild colic over 3 days. This colic had acutely exacerbated and was unresponsive to analgesic treatment, and was referred to Massey University Veterinary Teaching Hospital. CLINICAL FINDINGS: On examination the heart rate was 100 beats per minute, and mucous membranes were pale and tacky. A large mass was detected on transrectal palpation in the caudal abdomen to the left of midline. Explorative laparotomy revealed severe haemoperitoneum and several masses that were associated with the reproductive tract. The mare was then subject to euthanasia. On post-mortem examination, adjacent and attached to each ovary were soft, lobulated dark red masses up to 200 mm in diameter. Similar masses were present in the omentum and on the peritoneal surface of the diaphragm and the serosa of the spleen and liver. Histopathology revealed that the neoplastic component of the masses comprised proliferating cuboidal to columnar cells forming disorganised acini and cords separated by dense collagenous stroma. Immunohistochemistry showed the neoplastic cells were positive for cytokeratin AE1/AE3 and vimentin, but negative for cytokeratin 7 and inhibin α. DIAGNOSIS: Bilateral ovarian adenocarcinoma with transcoelomic metastasis and terminal decompensation due to rupture of a neoplastic mass and consequent haemoperitoneum. CLINICAL RELEVANCE: To the authors' knowledge, bilateral ovarian adenocarcinoma has not been previously reported in a horse. Ovarian adenocarcinoma should be considered when horses present with haemoperitoneum and colic. Further research is required on the immunohistochemical differentiation of adenocarcinoma of ovarian and intestinal origin in the horse.

A retrospective study of post-mortem examination findings in takahe (Porphyrio hochstetteri).

AIMS: To identify the major diseases or disease processes affecting both captive and wild populations of takahe (Porphyrio hochstetteri) in birds submitted for post-mortem examination between 1992 and 2007, and to survey archived wild dead takahe for the presence of Erysipelothrix rhusiopathiae. METHODS: Reports of 199 post-mortem examinations submitted between 1992 and 2007 were reviewed retrospectively. The reports comprised 56 eggs, 51 chicks up to 6 months of age, 13 sub-adults 6-18 months old, 74 adults and five birds where age was not recorded. Bone marrow flushed from the keel of 34 frozen adult takahe was assayed for the presence of E. rhusiopathiae, using PCR analysis. RESULTS: Of the eggs examined, 6/56 (11%) had no recorded diagnosis, 24/56 (43%) were infertile and 26/56 (46%) showed embryonic mortality at various stages. Excluding eggs, the cause of death could not be determined in 64/138 (46%) birds with a recorded age that were examined. Contributing factors for the low rate of diagnosis included advanced decomposition of many carcasses, the freezing of some birds prior to pathological investigation, long delays between recovery and submission for post-mortem examination, and variation in the extent of post-mortem examination and ancillary diagnostic testing. Common post-mortem examination findings in chicks included those related to cardiac disease [11/51 (22%)], infectious or inflammatory disease [9/51 (18%)] and trauma [7/51 (14%)]. In adult birds, the most common post-mortem examination findings were due to infectious or inflammatory disease [15/74 (20%)] including septicaemia due to E. rhusiopathiae [3/74 (7%)], followed by degenerative conditions due to diet, age and husbandry [9/74 (12%)]. Bacteria implicated in the causes of death included Escherichia coli, E. rhusiopathiae, Salmonella enterica serovar Typhimurium, and Pseudomonas aeruginosa. Only 1/34 (3%) of the frozen carcass examined using PCR was positive for E. rhusiopathiae. CONCLUSIONS: This study highlights a need for improved recovery, storage and submission of dead takahe for pathological examination; consistency in post-mortem examination, ancillary testing and recording of findings; and finally, regular communication between wildlife pathologists, conservation workers and representatives of Ngai Tahu.

PCBs and organochlorine pesticides in Hector's (Cephalorhynchus hectori hectori) and Maui's (Cephalorhynchus hectori maui) dolphins.

Polychlorinated biphenyls (PCBs) and organochlorine (OC) pesticide levels were determined in blubber samples collected from stranded and incidentally by-caught Hector's (Cephalorhynchus hectori hectori) and Maui's (Cephalorhynchus hectori maui) dolphins from New Zealand waters between 1997 and 2009. PCBs (45 congeners) and a range of OC pesticides including dieldrin, hexachlorocyclohexane (HCH) and dichlorodiphenyltrichloroethane (DDT), along with its metabolites DDE and DDD were determined. OC pesticides dieldrin, p,p'-DDE, p,p'-DDD and p,p'-DDT were present at the highest concentrations. Sum DDT concentrations ranged from 93.7 to 8210 (Mean=1358, S.D=1974) and 252.4 to 57,390 (Mean=12,389, S.D=18,161)microg/kg wet weight in females and males, respectively. Similarly, Sigma45CB concentrations ranged from 45.5 to 981.3 (Mean=333.2, S.D=265.8) and 60.5 to 5574 (Mean=1833, S.D=1659)microg/kg wet weight in females and males, respectively. The transfer of SigmaDDTs and summed PCBs (both as SigmaICES7CBs and Sigma45CBs) between a pregnant female and her unborn fetus was calculated at 5.7% and 4.3%, respectively. As the fetus was close to term, this likely represents the degree of placental transfer. Concentrations of OC pesticides determined in the present study are higher than those previously reported for Hector's dolphins. Sum DDT and DDE/SigmaDDT levels calculated reveal New Zealand's legacy of DDT usage, particularly off the east coast of the South Island.

Microphthalmia in Texel lambs.

AIM: To describe an outbreak of congenital microphthalmia in Texel lambs in New Zealand. METHODS: Affected eyes were fixed in 10% formol saline, processed routinely for histopathology and stained with haematoxylin and eosin, Holme's silver and haematoxylin with and without luxol fast blue, and luxol fast blue alone. RESULTS: Macroscopically, the globes and optic nerves were hypoplastic. Histologically, all embryonic components of the eye were present, but the lens, ciliary body, iris and retina were dysplastic. Axons of the hypoplastic optic nerve were poorly myelinated. CONCLUSIONS: Congenital microphthalmia occurs in Texel sheep in New Zealand, and is likely inherited as an autosomal recessive trait.

Squamous cell carcinoma in a tuatara (Sphenodon punctatus).

CASE HISTORY: A mature male tuatara was presented with a history of recurrent cloacal prolapse. CLINICAL AND PATHOLOGICAL FINDINGS: The prolapsed tissue included a 12 mm diameter mass, which histologically showed cords and nodules of neoplastic epithelial cells. DIAGNOSIS: The mass was diagnosed as a squamous cell carcinoma with ulceration of the overlying stratified squamous epithelium and diffuse inflammation of the surrounding dermis. CLINICAL RELEVANCE: This case presented a rare opportunity to investigate squamous cell carcinoma in a long-lived lower vertebrate species. Future recurrence or metastasis of the mass may add useful information to the current base of knowledge of the behaviour of malignant neoplasms in reptiles.