Evaluation of parameters of health in Australian fur seal (Arctocephalus pusillus doriferus) pups during early and mid-lactation

The physiological and behavioural development of diving was examined in Australian fur seal (Arctocephalus pusillus doriferus) pups to assess whether animals at weaning are capable of exploiting the same resources as adult females. Haematocrit, haemoglobin and myoglobin contents all increased throughout pup development though total body oxygen stores reached only 71% of adult female levels just prior to weaning. Oxygen storage components, however, did not develop at the same pace. Whereas blood oxygen stores had reached adult female levels by 9 months of age, muscle oxygen stores were slower to develop, reaching only 23% of adult levels by this age. Increases in diving behaviour corresponded to the physiological changes observed. Pups spent little time (<8%) in the water prior to moulting (age 1-2 months) whereas following the moult, they spent >27% of time in the water and made mid-water dives (maximum depth 35.7 +/- 2.9 m) with durations of 0.35 +/- 0.03 min. By 9 months (just prior to weaning), 30.5 +/- 9.3% of all dives performed were U-shaped benthic dives (maximum depth 65.0 +/- 6.0 m) with mean durations of 0.87 +/- 0.25 min, significantly shorter than those of adult females. These results suggest that while Australian fur seal pups approaching the age of weaning are able to reach similar depths as adult females, they do not have the physiological capacity to remain at these depths for sufficient durations to exploit them to the same efficiency.

Current prevalence of adult Uncinaria spp. in northern fur seal ( Callorhinus ursinus) and California sea lion ( Zalophus californianus) pups on San Miguel Island, California, with notes on the biology of these hookworms

Fur seal populations in the Southern Hemisphere were plundered in the late 1700s and early 1800s to provide fur for a clothing industry. Millions of seals were killed resulting in potentially major ecosystem changes across the Southern Hemisphere, the consequences of which are unknown today. Following more than a century of population suppression, partly through on-going harvesting, many of the fur seal populations started to recover in the late 1900s. Australian fur seals (Arctocephalus pusillus doriferus), one of the most geographically constrained fur seal species, followed this trend. From the 1940s to 1986, pup production remained at approximately 10,000 per year, then significant growth commenced. By 2007, live pup abundance had recovered to approximately 21,400 per year and recovery was expected to continue However, a species-wide survey in 2013 recorded a 20% decline, to approximately 16,500 live pups. It was not known if this decline was due to 2013 being a poor breeding year or a true population reduction. Here we report the results of a population-wide survey conducted in 2017 and annual monitoring at the most productive colony, Seal Rocks, Victoria that recorded a large decline in live pup abundance (-28%). Sustained lower pup numbers at Seal Rocks from annual counts between 2012–2017 (mean = 2908 ± 372 SD), as well as the population-wide estimate of 16,903 live pups in 2017, suggest that the pup numbers for the total population have remained at the lower level observed in 2013 and that the 5-yearly census results are not anomalies or representative of poor breeding seasons. Potential reasons for the decline, which did not occur range-wide but predominantly in the most populated and long-standing breeding sites, are discussed. To enhance adaptive management of this species, methods for future monitoring of the population are also presented. Australian fur seals occupy several distinct regions influenced by different currents and upwellings: range-wide pup abundance monitoring enables comparisons of ecosystem status across these regions. Forces driving change in Australian fur seal pup numbers are likely to play across other marine ecosystems, particularly in the Southern Hemisphere where most fur seals live.

The Australian fur seal (Arctocephalus pusillus doriferus) has experienced a slower than expected population recovery since the end of the commercial sealing era, with a high third trimester abortion rate. There is currently no known proximate cause. Coxiella burnetii (Cb) is a well-known cause of abortion in domestic and wildlife species and an important zoonotic pathogen. It has been recorded from a small number of northern hemisphere marine mammals and may be a potential contributory factor to decreasing populations of northern fur seals (Callorhinus ursinus) and Steller sealions (Eumetopias jubatus). It has not been recorded from marine mammals in the southern hemisphere but is well documented in ruminants and wildlife in Australia as a cause of reproductive failure. Third trimester aborted fetuses (n = 46) and full-term placentas (n = 66) from Australian fur seals, were collected on Kanowna Island and Seal Rocks in Bass Strait, south-eastern Australia. Utilizing routine hematoxylin and eosin histopathology, Cb immunohistochemistry and two different qPCR targets–htpAB and com1, Cb was identified. Routine histopathology and immunohistochemistry were insensitive for the detection of Cb. The detected Cb prevalence ranged from 10.6% for com1 up to 40.9% with htpAB. Coxiella burnetti was readily detected in full-term placentas but in aborted fetal material only in a single placenta associated with a still birth. The exact significance is currently unclear, but this highlights that Cb is present in Australian fur seals, breeding in Bass Strait. Bass Strait is in one of the world’s fastest warming oceanic regions and marine mammals breeding in the area are likely to be key indicators of marine ecosystem stressors. This first description of Cb in a marine mammal from the southern hemisphere, highlights the need to further investigate the potential risks this pathogen poses to Australian fur seals and sympatric marine mammals. Additionally, it is important to determine the zoonotic risk of this pathogen to persons working with, and in proximity of, Australian fur seal breeding colonies.

Australian fur seals (Arctocephalus pusillus doriferus) breed on Bass Strait islands in Victoria and Tasmania. They have been recorded in South Australia (SA) for many years as non-breeding visitors and on Kangaroo Island frequently since 1988, mostly in breeding colonies of the New Zealand fur seal (A. forsteri) which is the most numerous pinniped in SA. Australian fur seals have displaced New Zealand fur seals from sections of the Cape Gantheaume colony on Kangaroo Island. North Casuarina Island produced 29 Australian fur seal pups in February 2008. Australian fur seal pups were larger than New Zealand fur seal pups in the same colony and have been identified genetically using a 263-bp fragment of the mitochondrial DNA control region. North Casuarina Island has been an important breeding colony of New Zealand fur seals, but pup numbers there decreased since 1992–93 (contrary to trends in SA for New Zealand fur seals), while numbers of Australian fur seals there have increased. This study confirms that Australian fur seals breed in SA. The two fur seal species compete for space onshore at several sites. Australian fur seals may compete for food with endangered Australian sea lions (Neophoca cinerea) because both are bottom feeders.

Anthropogenic activities and pollution are impacting marine environments globally. This has resulted in increasing numbers of human-associated phylotypes of Escherichia coli, an indicator of faecal contamination, being found in both aquatic environments and upper trophic marine mammals that are considered sentinels for marine health. The objective of this study was to determine the presence and diversity of E. coli in pups of three species of free-ranging pinnipeds in Australia. Faecal samples (n = 964) were collected between 2016 and 2019 from Australian sea lion (Neophoca cinerea), Australian fur seal (Arctocephalus pusillus doriferus) and long-nosed fur seal (Arctocephalus forsteri) pups from eight breeding colonies extending along the Southern Australian coast. E. coli were isolated from 842 (87.3%) samples and molecular screening was applied to assign isolates to E. coli phylotypes and sub-types. The human associated E. coli phylotype B2 was the most frequently isolated in all species at seven of the eight colonies, with 73.7% of all E. coli isolates belonging to this phylotype. Phylotype distribution did not differ significantly within or across species, breeding colonies or breeding seasons. Further analysis of B2 isolates into sub-types showed a significant difference in sub-type distribution across breeding seasons at two colonies (Seal Rocks and Cape Gantheaume). The predominance of the B2 phylotype could indicate that all colonies are exposed to similar levels of anthropogenic pollution. This widespread occurrence of the human-associated E. coli phylotypes highlights the imperative for ongoing monitoring and surveillance of microbes in both the marine environment and sentinel species.

The peritoneal cavity (PNC) and intestine of northern fur seal (Callorhinus ursinus) pups and California sea lion (Zalophus californianus) pups that died in late July and early August, 2003, on San Miguel Island, California, were examined for hookworms. Prevalence and morphometric studies were done with the hookworms in addition to molecular characterization. Based on this and previous molecular studies, hookworms from fur seals are designated as Uncinaria lucasi and the species from sea lions as Uncinaria species A. Adult hookworms were found in the PNC of 35 of 57 (61.4%) fur seal pups and of 13 of 104 (12.5%) sea lion pups. The number of hookworms located in the PNC ranged from 1 to 33 (median = 3) for the infected fur seal pups and 1 to 16 (median = 2) for the infected sea lion pups. In addition to the PNC, intestines of 43 fur seal and 32 sea lion pups were examined. All of these pups were positive for adult hookworms. The worms were counted from all but one of the sea lion pups. Numbers of these parasites in the intestine varied from 3 to 2,344 (median = 931) for the fur seal pups and 39 to 2,766 (median = 643) for the sea lion pups. Sea lion pups with peritoneal infections had higher intensity infections in the intestines than did pups without peritoneal infections, lending some support for the hypothesis that peritoneal infections result from high-intensity infections of adult worms. There was no difference in intestinal infection intensities between fur seal pups with and without peritoneal infections. Female adult hookworms in the intestines of both host species were significantly larger than males, and sea lion hookworms were larger than those in fur seals. Worms in the intestine also were larger than worms found in the PNC. Gene sequencing and (RFLP) analysis of (PCR) amplified (ITS) ribosomal DNA were used to diagnose the species of 172 hookworms recovered from the PNC and intestine of 18 C. ursinus and seven Z. californianus hosts. These molecular data revealed that U. lucasi (hookworm of C. ursinus) and Uncinaria species A (of Z. californianus) infrequently mature in the intestine of the opposite host species in California rookeries. However, there is no support from molecular data for the hypothesis that cross-infection with "the wrong" Uncinaria species is a contributing factor in these cases of host peritonitis. The major significance of this research is the unusual finding of adult hookworms in the PNC of so many dead pups. No obvious explanation for this occurrence could be determined. Further research, like in the present study, should help understand and monitor the apparent ever changing role of hookworm disease in the health of northern fur seal and California sea lion pups on SMI.

Context Coxiella burnetii is suspected as a novel pathogen contributing to decreased pup production in Australian fur seals (Arctocephalus pusillus doriferus). It has recently been described from a single breeding colony in Bass Strait, has previously been associated with two decreasing populations of northern hemisphere pinnipeds and is a known reproductive pathogen. Data around its disease ecology in marine mammals are sparse. Aims To determine whether environmental DNA (eDNA) can be used to survey for C. burnetii in Australian fur seal breeding colonies. To determine whether C. burnetii in Australian fur seals is the same genotype as terrestrial Australian C. burnetii. Methods Soil samples were collected from Kanowna Island and Seal Rocks. Placental samples were collected from Kanowna Island. Soil was evaluated for eDNA using a quantitative polymerase chain reaction (qPCR) for com1 gene. Placental samples were evaluated with com1, htpAB and IS1111 markers. Multiple-locus variable number of tandem repeats analysis for three microsatellite loci (ms-24, ms-28 and ms-33) was used to determine relatedness to Australian C. burnetii genotypes. Key results eDNA results varied between pre-and post-pupping at Seal Rocks. When targeting the com1 gene, the post-pupping prevalence at Kanowna Island and Seal Rocks was 59.6% and 90%, respectively. eDNA PCR inhibition of samples was low at 1.9%. There was very poor, sporadic to absent IS1111 amplification in placental samples. The com1 and htpAB qPCRs had an overall prevalence across placental samples of 39.2% and 56.7% respectively. In 90.1% of placental samples (n = 11), the ms-28 locus amplified. Neither ms-24 nor ms-33 amplified. Conclusions eDNA is an effective tool to survey Australian fur seal breeding colonies in the post-pupping period for C. burnetii. The prevalence appears to be much higher in the Seal Rocks colony than in the Kanowna Island colony. It appears that this is not a terrestrial Australian genotype but rather closely related to genotypes detected in marine mammals in the northern hemisphere. Implications This research significantly expands our ability to survey for C. burnetii in Australian fur seals and other marine mammals. It highlights knowledge gaps in our understanding of the disease ecology and phylogeny of C. burnetii in marine mammals.

The development of cardiac control in association with terrestrial respiration patterns was examined throughout the period of maternal dependence in Australian fur seal pups. Resting eupnoic heart rate and respiration rate were significantly correlated (r (2) = 0.49) and both decreased with age (P < 0.05 in both cases). From an early age (1 month), pups displayed terrestrial apnoeas (18.1 +/- 0.5 s) accompanied by substantial bradycardia (127 beats min(-1), a 13% decrease from eupnoic HR). Terrestrial apnoea duration increased significantly with age reaching a mean of 41 s just prior to weaning, slightly lower than the mean dive duration (52 s) previously recorded for pups of the same age. Correspondingly, mean apnoic heart rate decreased with age to 74 beats min(-1) just prior to weaning, representing a 25% decrease on eupnoic heart rate. Importantly, concomitant with the decrease in mean apnoic heart rate with age, an increase in the control of bradycardia was evident with the variability in instantaneous apnoic heart decreasing such that older pups were able to maintain a low steady heart rate for the duration of the apnoea. The changes seen in these parameters are similar to those reported during postnatal development in elephant seals (Mirounga spp.) and harbour seals (Phoca vitulina), and are considered indicative of the development of cardiac control. These findings suggest a common strategy for the development of bradycardia control in both otariid and phocid seals.

Mass and length growth models were determined for male (n = 69) and female (n = 163) Australian fur seals (Arctocephalus pusillus doriferus) collected at a breeding colony on Seal Rocks (38˚31′S, 145˚06′E), Bass Strait, in south-east Australia, between February and November during 1970–72. Growth was best described by the logistic model in males and the von Bertalanffy model in females. Asymptotic mass and length were 229 kg and 221 cm for males, and 85 kg and 163 cm for females. In all, 95% of asymptotic mass and length were attained by 11 years and 11 years, respectively, in males compared with 9 years and 5 years, respectively, in females. Males grew in length faster than females and experienced a growth spurt in mass coinciding with the onset of puberty (4–5 years). The onset of puberty in females occurs when approximately 86% of asymptotic length is attained. The rate of growth and sexual development in Australian fur seals is similar to (if not faster than) that in the conspecific Cape fur seal (A. p. pusillus), which inhabits the nutrient-rich Benguela current. This suggests that the low marine productivity of Bass Strait may not be cause of the slow rate of recovery of the Australian fur seal population following the severe over-exploitation of the commercial sealing era. Sternal blubber depth was positively correlated in adult animals with a body condition index derived from the residuals of the mass–length relationship (males: r2 = 0.38, n = 19, P &lt; 0.001; females: r2 = 0.22, n = 92, P &lt; 0.001), confirming the validity of using such indices on otariids. Sternal blubber depth varied significantly with season in adult animals. In males it was lowest in winter and increased during spring prior to the breeding season (r2 = 0.39, n = 19, P &lt; 0.03) whereas in females it was greatest during winter (r2 = 0.05, n = 122, P&lt; 0.05).

The behaviours used by mammalian predators to capture and process their prey play a pivotal role in determining hunting success when targeting different species, or prey in different foraging environments. This is especially true in aquatic environments, where marine mammals like pinnipeds encounter a wide range of prey species while hunting both in open water and near the sea floor. However, due to difficulties faced when observing behaviour in diving mammals, little is known about the mechanics of prey handling in most pinnipeds species. The aim of this thesis was to remedy this by performing captive feeding trials that explored how foraging behaviours varied under different conditions for otariid species that have not been studied before now. When capturing small fish in open water, Australian fur seals (Arctocephalus pusillus doriferus) used raptorial biting. Rather than simply snapping at prey with the jaws, this involved combined use of biting and suction, where they used suction to draw prey within range of the teeth before the jaws snapped shut. In contrast, when capturing prey concealed within a feeding device, fur seals used strong suction alone to draw prey into the mouth. Australian and subantarctic fur seals (Arctocephalus tropicalis) swallowed small prey whole after the initial capture, while large prey was first brought to the surface and processed into smaller pieces by shaking. By chewing on prey between shakes, the fur seals created weak points that made it more likely to break when shaken further. Australian sea lions (Neophoca cinerea) were also able to chew and shake food; however, unlike fur seals, they also tore food held between their forelimbs and teeth. By directly observing how prey capture and processing occurs in otariid pinnipeds, we are able to begin to draw broader comparisons between these species and the other main clades of pinnipeds. Phocine phocids (the northern seals) also use biting and suction when capturing large vs. small prey, but unlike fur seals they are able to grip prey in clawed forelimbs while processing it with their teeth. Monachine phocids (the “southern” seals) have forelimbs intermediate in morphology between the clawed phocines and highly derived otariids. Those that have been studied or observed seem to display similar behaviours to the fur seals from our studies. Australian sea lions are unusual in being similar to phocine seals that hold prey in their forelimb flippers, but rather than using claws to secure prey, they squeezed prey between flat palms. These patterns highlight key differences in behaviour that may relate to differences in forelimb morphology that stem from the degrees to which each clade has adapted its anatomy for aquatic locomotion. By making direct observations of behaviour for a wide range of pinniped species we can therefore identify patterns that provide a better understanding of the role that foraging behaviour played throughout pinniped evolution. These results also provide insights into how prey capture and processing behaviours influence the foraging choices made by seals hunting at sea, providing insights into what makes pinnipeds such successful marine predators.

Observing how pinnipeds respond to variations in climatic and oceanographic conditions informs marine managers on factors that could limit their range, foraging ability and breeding success. Here, we examine how Australian fur seals (Arctocephalus pusillus doriferus) at Seal Rocks, Victoria, Australia, responded to normal climatic conditions from August 2009 to January 2010, which included their Austral spring‐summer breeding period, to investigate their tolerances to a range of environmental stimuli. Seal numbers ashore and a range of climatic variables were collected hourly during daylight periods and compared using Generalized Additive Mixed Models (GAMMs). Air temperature was the most consistent predictor of haul‐out behavior, with seal numbers ashore declining as air temperature increased (effect size −50%, edf 1.00, P &lt; 0.001). Increased wave height (effect size 74%, edf 1.00, P &lt; 0.001) and wind speed (effect size 79%, edf 1.00, P &lt; 0.001) were associated with increased seal numbers ashore. Potentially, higher air temperatures reduce the seals tolerance to remain out of the water, while high wind/wave action increases at‐sea metabolic costs. These results demonstrate how changes in climate could alter a seal's ability to remain ashore, to rest or breed, and its ability to forage effectively, thus driving changes in population status and range.

Australian fur seals (Arctocephalus pusillus doriferus) are conspicuous, top-level predators in coastal waters of south-eastern Australia that were over-harvested during the 1800s and have had a delayed recovery. A previous species-wide estimate of live pups in 2002 recorded a near-doubling of annual pup production and a 5% annual growth rate since the 1980s. To determine if pup production increased after 2002, we estimated live pup numbers in 2007. Pups were recorded at 20 locations: 10 previously known colonies, three newly recognised colonies and seven haul-out sites where pups are occasionally born. Two colonies adjacent to the Victorian coast accounted for 51% of live pups estimated: Seal Rocks (5660 pups, 25.9%) and Lady Julia Percy Island (5574 pups, 25.5%). Although some colonies were up and some were down in pup numbers, the 2007 total of 21 882 ± 187 (s.e.) live pups did not differ significantly from a recalculated estimate of 21 545 ± 184 in 2002, suggesting little change to overall population size. However, the colonisation of three new sites between 2002 and 2007 indicates population recovery has continued.

The intracellular pathogens Toxoplasma gondii, Brucella spp., and Chlamydia spp. are all known causative agents of abortion in wildlife. Both T. gondii and Brucella spp. have been identified in marine mammal abortions and a limited number of studies have detected their potential presence in Australian fur seals (Arctocephalus pusillus doriferus), but data are sparse for these pathogens in Australian fur seal breeding colonies. Australian fur seals have been shown to have a high degree of third-trimester pregnancy loss in one of their largest breeding colonies. Additionally, pup production has declined at the largest breeding colony for the species. This study surveyed the presence of T. gondii, Brucella spp., and Chlamydia spp. as potential infectious causes of this reproductive loss. Aborted fetuses were collected from two of the largest breeding colonies for the species, Seal Rocks (n=19) and Kanowna Island (n=34). These were examined grossly and through histopathological evaluation, in conjunction with molecular testing for all three pathogens. Placentas were collected from full-term births during the pupping season from Kanowna Island (n=118). These were used to compare the molecular prevalence of the three pathogens in presumed successful pregnancies. Chlamydia spp. was not detected in aborted fetuses in this study. Brucella spp. was detected with PCR in both aborted fetuses (9.4%) and placentas from full-term births (3.4%), and T. gondii was detected using routine histopathology (n=2/53), immunohistochemistry (n=3/4), and PCR (n=4/53) in tissues from aborted fetuses. Toxoplasma gondii was present in 7.5% of third-trimester abortions and absent from all full-term placentas. Brucella spp. was detected in both aborted fetuses and full-term placentas. This is the first description of vertical transmission of T. gondii in a marine mammal from the southern hemisphere.

Vessel traffic is the greatest contributor to marine anthropogenic noise pollution, and particularly affects species utilising coastal areas. Seal Rocks (Victoria), the largest Australian fur seal breeding colony, has a relatively small vessel exclusion zone during the pupping and breeding season, when vessel traffic is at its peak. It is necessary to understand the impacts of visitation at sensitive marine sites to ensure they are adequately protected from human disturbance. This study assessed the behavioural response of hauled-out Australian fur seals at Seal Rocks to controlled vessel noise exposure during peak and off-peak vessel visitation periods. High level noise exposure (76–80 dB) resulted in increased vigilance and interruption to vital behaviours in both peak and off-peak periods. These behavioural responses were limited to the period of noise exposure but are nevertheless indicative of disturbance. It is important to ensure regulations intended to protect sensitive sites are up-to-date and evidence based.