Mycobacterium pinnipedii causes tuberculosis in a number of pinniped species, and transmission to cattle and humans has been reported. The aims of this study were to: characterize the pathology and prevalence of tuberculosis in New Zealand marine mammals; use molecular diagnostic methods to confirm and type the causal agent; and to explore relationships between type and host characteristics. Tuberculosis was diagnosed in 30 pinnipeds and one cetacean. Most affected pinnipeds had involvement of the pulmonary system, supporting inhalation as the most common route of infection, although ingestion was a possible route in the cetacean. PCR for the RD2 gene confirmed M . pinnipedii as the causal agent in 23/31 (74%) cases (22 using DNA from cultured organisms, and one using DNA from formalin-fixed paraffin-embedded (FFPE) tissue), including the first published report in a cetacean. RD2 PCR results were compared for 22 cases where both cultured organisms and FFPE tissues were available, with successful identification of M . pinnipedii in 7/22 (31.8%). In cases with moderate to large numbers of acid-fast bacilli, RD2 PCR on FFPE tissue provided a rapid, inexpensive method for confirming M . pinnipedii infection without the need for culture. VNTR typing distinguished New Zealand M . pinnipedii isolates from M . pinnipedii isolated from Australian pinnipeds and from common types of M . bovis in New Zealand. Most (16/18) M . pinnipedii isolates from New Zealand sea lions were one of two common VNTR types whereas the cetacean isolate was a type detected previously in New Zealand cattle.

CASE HISTORY A little penguin (Eudyptula minor) of wild origin, in captivity at Wellington Zoo, became inappetent and lethargic in March 2013. Despite supportive care in the zoo's wildlife hospital, the bird died within 24 hours. CLINICAL FINDINGS Weight loss, dehydration, pale mucous membranes, weakness, increased respiratory effort and biliverdinuria were apparent on physical examination. Microscopic evaluation of blood smears revealed intra-erythrocytic stages of Plasmodium spp. and a regenerative reticulocytosis in the absence of anaemia. PATHOLOGICAL FINDINGS Post-mortem findings included reduced body condition, dehydration, pulmonary congestion and oedema, hepatomegaly, splenomegaly, hydropericardium and subcutaneous oedema. Histopathological findings included protozoal organisms in sections of lung, liver and spleen. A marked, diffuse, sub-acute interstitial histiocytic pneumonia was present. Accumulation of haemosiderin was noted in the Kupffer cells of the liver and in histiocytic-type cells in the spleen. MOLECULAR TESTING DNA was extracted from frozen portions of the liver. Nested PCR results and DNA sequencing confirmed infection of the deceased little penguin with Plasmodium (Huffia) elongatum lineage GRW06. DIAGNOSIS Avian malaria due to Plasmodium (Huffia) elongatum GRW06 RETROSPECTIVE INVESTIGATION A retrospective analysis of 294 little penguin cases in the Massey University post-mortem database revealed three other potential avian malaria cases. Analysis of archived tissues using a nested PCR for Plasmodium spp. followed by DNA sequencing revealed that a little penguin which died at Auckland Zoo was infected with P. elongatum GRW06 and two wild little penguins found dead on New Zealand beaches were infected with P. relictum SGS1 and Plasmodium. sp. lineage LINN1. Therefore, the overall frequency of deaths in little penguins associated with avian malaria was 4/295 (1.36%). CLINICAL RELEVANCE Our results suggest that avian malaria is associated with sporadic mortality in New Zealand's little penguins both in the wild and in captivity, but there is no evidence of mass mortality events due to Plasmodium spp. infection.

The relative importance of intrinsic and extrinsic determinants of animal foraging is often difficult to quantify. The most southerly breeding mammal, the Weddell seal, remains in the Antarctic pack-ice year-round. We compared Weddell seals tagged at three geographically and hydrographically distinct locations in East Antarctica (Prydz Bay, Terre Adélie, and the Ross Sea) to quantify the role of individual variability and habitat structure in winter foraging behaviour. Most Weddell seals remained in relatively small areas close to the coast throughout the winter, but some dispersed widely. Individual utilisation distributions (UDi, a measure of the total area used by an individual seal) ranged from 125 to 20,825 km2. This variability was not due to size or sex but may be due to other intrinsic states for example reproductive condition or personality. The type of foraging (benthic vs. pelagic) varied from 56.6 ± 14.9% benthic dives in Prydz Bay through 42.1 ± 9.4% Terre Adélie to only 25.1 ± 8.7% in the Ross Sea reflecting regional hydrographic structure. The probability of benthic diving was less likely the deeper the ocean. Ocean topography was also influential at the population level; seals from Terre Adélie, with its relatively narrow continental shelf, had a core (50%) UD of only 200 km2, considerably smaller than the Ross Sea (1650 km2) and Prydz Bay (1700 km2). Sea ice concentration had little influence on the time the seals spent in shallow coastal waters, but in deeper offshore water they used areas of higher ice concentration. Marine Protected Areas (MPAs) in the Ross Sea encompass all the observed Weddell seal habitat, and future MPAs that include the Antarctic continental shelf are likely to effectively protect key Weddell seal habitat.

Graphical Abstract

Albatrosses are among the world's most imperiled vertebrates, with 73% of species threatened with extinction. Ingestion of plastic is a well-recognized threat among three North Pacific species, but lesser known in the southern hemisphere, where it is considered a minor threat. As plastic entering the ocean is increasing while albatross populations decline, the threat of ocean plastic to albatross populations may be underestimated. We present case studies of 107 beach-cast albatrosses of twelve species, received by wildlife hospitals in Australia and New Zealand, and estimate plastic ingestion and mortality rates for albatrosses in the southern hemisphere. Ingested plastic was present in 5.6% of individuals, and the cause of death in half of these cases. We estimate ingestion of plastic may cause 3.4-17.5% of nearshore mortalities and is worth consideration as a substantial threat to albatross populations. We provide clinical findings and "checklist" methodologies for identifying potential cases of foreign-body gastrointestinal obstruction. We suggest practical policy responses, empowering decision makers to reduce albatross mortality from anthropogenic sources.