A novel approach identified the FOLR1 gene, a putative regulator of milk protein synthesis

This study has utilised comparative functional genomics to exploit animal models with extreme adaptation to lactation to identify candidate genes that specifically regulate protein synthesis in the cow mammary gland. Increasing milk protein production is valuable to the dairy industry. The lactation strategies of both the Cape fur seal (Artocephalus pusillus pusillus) and the tammar wallaby (Macropus eugenii) include periods of high rates of milk protein synthesis during an established lactation and therefore offer unique models to target genes that specifically regulate milk protein synthesis. Global changes in mammary gene expression in the Cape fur seal, tammar wallaby, and the cow (Bos taurus) were assessed using microarray analysis. The folate receptor α (FOLR1) showed the greatest change in gene expression in all three species [cow 12.7-fold (n = 3), fur seal 15.4-fold (n = 1), tammar 2.4-fold (n = 4)] at periods of increased milk protein production. This compliments previous reports that folate is important for milk protein synthesis and suggests FOLR1 may be a key regulatory point of folate metabolism for milk protein synthesis within mammary epithelial cells (lactocytes). These data may have important implications for the dairy industry to develop strategies to increase milk protein production in cows. This study illustrates the potential of comparative genomics to target genes of interest to the scientific community.     

Behavior of Cape fur seals (Arctocephalus pusillus pusillus) in relation to temporal variation in predation risk by white sharks (Carcharodon carcharias) around a seal rookery in False Bay,South Africa

The marked differences in predation risk posed by white sharks (Carcarodon carcarias) at island rookeries of Cape fur seals (Arctocephalus pusillus pusillus) offer a quasi‐experimental design within a natural system for exploring how prey adjust their behavior in response to temporal variation in predation risk. Here we compare movement of juvenile and adult Cape fur seals at a high risk (Seal Island) and low risk (Egg Island) rookery. We further compare juveniles and adults at Seal Island in low and high risk seasons and at low and high risk times of day within those seasons. Adult fur seals at Seal Island avoided traversing the zone of high white shark predation risk during the high risk period (0700–0959) in the season of high risk (winter), but not during the low risk season (summer). By contrast, adult fur seals at Egg Island showed no temporal discretion in either season. Unlike juvenile fur seals at Egg Island, juveniles at Seal Island adjusted their temporal movement patterns to more closely mimic adult seal movement patterns. This suggests that exposure to predators is the primary driver of temporal adjustments to movement by prey species commuting from a central place.     

Killing of Cape fur seal (Arctocephalus pusillus pusillus) pups by brown hyenas (Parahyaena brunnea) at mainland breeding colonies along the coastal Namib Desert

Brown hyenas (Parahyaena brunnea) scavenge and kill seal pups at mainland Cape fur seal (Arctocephalus pusillus pusillus) colonies. The prey encounter interval and interval between kills depended on seal density, and increased density resulted in an increase of the capture rate and increase in hunting efficiency from 14% in November to 47% in January. The time brown hyenas spent at the seal colony decreased with increasing seal density and increasing air temperatures. Nevertheless, they were regularly active during the day when less adult seals were present at the colony, which indicates that the attendance of adult seals might play a role in the choice of foraging time. Brown hyenas killed seal pups throughout the study period. The predation rate was independent of the availability of non-violent mortalities, but the absolute number of kills was positively density-dependent. Mass kill events were recorded throughout the study period and are therefore not unusual occurrences. The overabundance of easy and vulnerable prey may lead to an over stimulus situation that triggers killing independent of the consumption of the prey or the hunger state.     

Behavior of Cape fur seals (Arctocephalus pusillus pusillus) in response to spatial variation in white shark (Carcharodon carcharias) predation risk

Foraging and predation risk are often separated at rookeries of marine central place foragers, thus offering an opportunity to gain insight into how predator‐avoidance shapes the behavior of prey. Here we compare the behavior of Cape fur seals (Arctocephalus pusillus pusillus) at two island rookeries with and without white shark (Carcharodon carcharias) predations, and assess seal behavior in relation to marked spatiotemporal variation in risk at the high‐risk site (Seal Island, South Africa). Our results show that seal behavior at the two sites is comparatively similar in summer, when predation risk is low at both sites, but not in winter. Compared to seals at the “low‐risk” site, seals at Seal Island avoided deep‐water habitat around the island at high risk times and restricted their use of this habitat in favor of safe, shallow waters when engaging in social and thermoregulatory behaviors. Seals increased their frequency of jostling, porpoising, and diving when moving through the danger zone and seals in groups were safer than single individuals. Overall, our results suggest that seal behavior around the high‐risk site is strongly affected by predation risk, and show this rookery to be an excellent predator‐prey system at which to evaluate long‐standing ecological hypotheses.     

The influence of extracellular matrix and prolactin on global gene expression profiles of primary bovine mammary epithelial cells in vitro

An in vitro bovine mammosphere model was characterized for use in lactational biology studies using a functional genomics approach. Primary bovine mammary epithelial cells cultured on a basement membrane, Matrigel, formed three-dimensional alveoli-like structures or mammospheres. Gene expression profiling during mammosphere formation by high-density microarray analysis indicated that mammospheres underwent similar molecular and cellular processes to developing alveoli in the mammary gland. Gene expression profiles indicated that genes involved in milk protein and fat biosynthesis were expressed, however, lactose biosynthesis may have been compromised. Investigation of factors influencing mammosphere formation revealed that extracellular matrix (ECM) was responsible for the initiation of this process and that prolactin (Prl) was necessary for high levels of milk protein expression. CSN3 (encoding κ-casein) was the most highly expressed casein gene, followed by CSN1S1 (encoding αS1-casein) and CSN2 (encoding β-casein). Eighteen Prl-responsive genes were identified, including CSN1S1 , SOCS2 and CSN2, however, expression of CSN3 was not significantly increased by Prl and CSN1S2 was not expressed at detectable levels in mammospheres. A number of novel Prl responsive genes were identified, including ECM components and genes involved in differentiation and apoptosis. This mammosphere model is a useful model system for functional genomics studies of certain aspects of dairy cattle lactation.     

The composition of Australian fur seal (Arctocephalus pusillus doriferus) milk throughout lactation.

The proximate chemical composition of milk was determined throughout lactation in the Australian fur seal (Arctocephalus pusillus doriferus Wood Jones), a temperate species with an 11-mo lactation period typical of most temperate otariids. Average lipid and protein contents were 42% and 10%, respectively, which is similar to that recorded for the polar otariid species but higher than reported in other temperate, long-lactation species. Milk composition, however, varied substantially throughout lactation, with lipid content rising from 30% soon after parturition to 50% at 230 d postpartum before decreasing to 45% toward weaning. The trend in water content was opposite to that of lipid, while protein content increased only slightly during lactation from 10% to 12%. The peak in lipid content occurred at approximately 70% of the way through lactation, similar to that found in the Australian sea lion and several phocid seal species. Significant positive relationships were found between milk lipid content and both the duration of the preceding foraging trip and body mass. A highly significant and close correlation was found between milk water and lipid contents such that gross energy content may be accurately predicted from water content alone. Similar predictive equations have been found for several other otariid and phocid species, but these appear to differ between phylogenetic groups.     

Vagrant elephant seal predation on Cape fur seal pups, Plettenberg Bay, South Africa

Vagrant southern elephant seals (Mirounga leonina) are occasionally sighted along the coast of South Africa and are known to feed primarily on fish and squid. Phocid seals are not known to predate on mammals making the events described here exceptional. This note describes the successful and failed attempts of a vagrant male southern elephant seal (M. leonina) to consume Cape fur seal (Arctocephalus pusillus pusillus) pups in Plettenberg Bay, South Africa. Observations were made by crew and passengers aboard a commercial whale-watching vessel during November 2012. This is the first account of elephant seals eating anything other than fish, squid and penguins and suggests considerable plasticity in prey choice dictated by environmental conditions.     

EVALUATION OF AN EFFECTIVE METHOD TO ESTIMATE AGE OF CAPE FUR SEALS USING GROUND TOOTH SECTIONS

Teeth of known-age Cape fur seals Arctocephalus pusillus pusillus were used to validate age estimated from ground sections. In the canines, dentine growth layer groups (GLGs) reflected age accurately but no reliable readings could be obtained from GLGs in the cementum. Upper canines were the most suitable for age estimation. By contrast, in the postcanines where the cementum is thicker, only GLGs in the cementum could be used for age determination, but not with the same accuracy as for dentine in the canines. Therefore, it is recommended that GLGs in the dentine be used to determine age in the Cape fur seal. However, pulp cavities in canines closed at about 13 yr and consequently GLGs in the cementum of the postcanines should be used where the pulp cavities of canines are closed. Accurate estimation of age is not possible from the dentine of older animals.     

Characterisation of proteins in the milk of fur seals

Milk protein composition was investigated throughout the lactation periods of the Australian fur seal (Arctocephalus pusillus doriferus) and Antarctic fur seal (Arctocephalus gazella). The mean protein content of the milk was found to be 10.9% and 10.6% respectively. The concentration of total protein did not change during lactation, although a decline in casein content of the milk in late lactation was apparent. Milk protein concentration during a foraging/suckling cycle of the Antarctic fur seal analysed at the time of arrival on shore, and 24 h and 72 h after arrival was 12.8%, 11.4% and 12.5% respectively. Re-feeding animals at 72 h resulted in a significant increase in milk protein content to 14.9%. Characterisation of milk protein by SDS-PAGE analysis revealed 5 casein and 10 major whey protein bands. Amino-terminal sequencing indicated that the majority of the whey fraction of the milk is beta-lactoglobulin (beta-LG). The limited amino acid sequence indicated 3 different beta-LGs were secreted in the milk. Subsequently, RT-PCR was used to extend the sequence of one of the beta-LGs and translation of the 464 bp fragment indicated that it shared 79% sequence identity with feline beta-LG II.     

The efficacy of culling seals seen preying on seabirds as a means of reducing seabird mortality

In the 2006/2007 breeding season of Cape gannets Morus capensis at Malgas Island, the removal of 61 Cape fur seals Arctocephalus pusillus pusillus that preyed on gannet fledglings when they left to sea significantly reduced the mortality rate of these fledglings. However, because seals learned to avoid the boat used for their removal, it was not possible to remove all the seals that killed gannet fledglings and some mortality continued. The seals inflicting the mortality were all sub-adult males, with an average age of <5 years. Sustained removal of these animals may reduce this feeding behaviour, which is at present having an adverse impact on several threatened seabirds in the Benguela ecosystem.     

Mammary transmission of caprine arthritis encephalitis virus: a 3D model for in vitro study

Transmission of Caprine Arthritis Encephalitis virus (CAEV) from the mother to offspring is principally mediated by infected cells from colostrum and milk. The infection of the dam is often sub-clinical, and results in increased cellularity of milk, sometimes exacerbated by bacterial co-infections. Although monocytes are the major viral host cells, several other cell types, including epithelial mammary cells, fibroblasts and endothelial cells show low levels of in vivo infection. In vitro, however, all phenotypes of mammary gland cells are individually highly sensitive to CAEV infection. This suggests that local mechanisms act to control viral expression. Our goal is to analyse the mechanisms regulating local virus infection, including the physiological status of the mammary gland and bacterial co-infections. In this work, we present the development of a model for the in vitro reconstitution of mammary gland tissue using 3D cultures in Matrigel. Mononuclear cells from the blood are added to the 3D cultures in vitro. In these experimental conditions, the mammary cells spontaneously organize into mammospheres. Blood leucocytes migrate into the culture gel, and localize particularly at the periphery of the mammospheres. Mammospheres were susceptible to infection in vitro by CAEV, as shown by a cytopathic effect and expression of late CAEV antigen p30. This model will allow the in vitro study of virus expression, transfer of infection to mammary gland cells and interactions between the mammary gland cells, infected monocytes and immunocompetent cells. It will allow the study of mechanisms participating in the control of passage of pathogens into milk, according to the physiological and CAEV-infection status of the animal, microenvironment and the presence of bacterial co-infections.     

SUCCESSFUL USE OF A TRANSLOCATION PROGRAM TO INVESTIGATE DIVING BEHAVIOR IN A MALE AUSTRALIAN FUR SEAL, ARCTOCEPHALUS PUSILLUS DORIFERUS

This study reports some of the first foraging behavior data collected for male fur seals. A nonbreeding male Australian fur seal, Arctocephalus pusillus doriferus , captured at a commercial salmon farm in southern Tasmania, Australia, was relocated 450 km from the site of capture. The animal was equipped with a geolocating time-depth recorder that recorded diving behavior and approximate location for the 14.4 d that it took the seal to travel down the east coast of Tasmania and be recaptured at the salmon farm. During its time at sea, the seal spent most of its time over the relatively shallow shelf waters. It spent 30% of its time ashore on a number of different haul-out sites. The deepest dive was 102 m and the maximum duration was 6.8 min. "Foraging" type dives made up 31.2% of the time at sea and had a median duration of 2.5 min and a median depth of 14 m. The seal performed these dives more commonly during the latter part of its time at sea, while it was on the east coast. Unlike other fur seal species studied to date, there was no evidence of a diurnal foraging pattern; it made dives at all times of the day and night.     

Activity Budgets of Captive Cape Fur Seals (Arctocephalus pusillus) Under a Training Regime

Ethograms and time budgets are crucial for the behavioral assessment of nonhuman animals in zoos, and they serve as references for welfare research. This study was conducted to obtain detailed time budgets of trained Cape fur seals (Arctocephalus pusillus) in captivity, to evaluate variations of these patterns, and to determine whether abnormal behaviors had been displayed. Behavioral data for 3 Cape fur seals in the Wroclaw Zoo were collected, and more than 300 observation hours (during a 12-month period) per individual were analyzed. The studied animals exhibited a diversified repertoire of natural behaviors with apparent seasonal and daily patterns, and they did not present stereotypic behaviors. Significant differences of interaction rates between individuals suggest more frequent affiliative interactions among related animals. The absence of stereotypic behaviors, good health of individuals, and the presence of diversified natural behaviors indicated relatively good welfare of Cape fur seals kept in the Wroclaw Zoo.     

Tuberculosis in a captive colony of pinnipeds

Tuberculosis was diagnosed in 10 of 16 otariid seals upon post mortem examination. The species involved were New Zealand fur seals (Arctocephalus forsteri), Australian sea lions (Neophoca cinerea) and an Australian fur seal (Arctocephalus pusillus doriferus). Five seals died, four as a direct result of mycobacterial infection. One seal died of unrelated disease. The remaining 10 animals were subsequently tuberculin tested and then killed and necropsied. Tuberculous lesions were seen in five. Gross pathological changes were most commonly seen in the respiratory system. However, a generalized infection, a case with lesions confined to the liver and draining lymph nodes, and a case with tuberculous meningitis also were seen. Histological lesions were characterized by spindle cell proliferation and necrosis without mineralization or giant cell formation. The mycobacteria isolated was identified as belonging to the Mycobacterium tuberculosis complex but it appeared to be unique. Intradermal tuberculin testing showed promise as a diagnostic aid; however, the results were not statistically significant. Circumstances suggest that the initial infection was present when the seals were captured from the wild.     

Australian Fur Seals (Arctocephalus pusillus doriferus) Use Raptorial Biting and Suction Feeding When Targeting Prey in Different Foraging Scenarios

Foraging behaviours used by two female Australian fur seals (Arctocephalus pusillus doriferus) were documented during controlled feeding trials. During these trials the seals were presented with prey either free-floating in open water or concealed within a mobile ball or a static box feeding device. When targeting free-floating prey both subjects primarily used raptorial biting in combination with suction, which was used to draw prey to within range of the teeth. When targeting prey concealed within either the mobile or static feeding device, the seals were able to use suction to draw out prey items that could not be reached by biting. Suction was followed by lateral water expulsion, where water drawn into the mouth along with the prey item was purged via the sides of the mouth. Vibrissae were used to explore the surface of the feeding devices, especially when locating the openings in which the prey items had been hidden. The mobile ball device was also manipulated by pushing it with the muzzle to knock out concealed prey, which was not possible when using the static feeding device. To knock prey out of this static device one seal used targeted bubble blowing, where a focused stream of bubbles was blown out of the nose into the openings in the device. Once captured in the jaws, prey items were manipulated and re-oriented using further mouth movements or chews so that they could be swallowed head first. While most items were swallowed whole underwater, some were instead taken to the surface and held in the teeth, while being vigorously shaken to break them into smaller pieces before swallowing. The behavioural flexibility displayed by Australian fur seals likely assists in capturing and consuming the extremely wide range of prey types that are targeted in the wild, during both benthic and epipelagic foraging.     

Natural mortality of puffadder shysharks due to Cape fur seals and black‐backed kelp gulls at Seal Island, South Africa

Natural mortality of puffadder shysharks Haploblepharus edwardsii due to two species of marine tetrapod, the Cape fur seal Arctocephalus pusillus pusillus and the black‐backed kelp gull Larus dominicanis vetula , is reported. These data constitute the first multiple observations of natural mortality of any cartilaginous fish due to object play or kleptoparasitism by marine tetrapods. Evidence of range extension of the puffadder shyshark to False Bay, South Africa is presented.     

Sharks shape the geometry of a selfish seal herd: experimental evidence from seal decoys

Many animals respond to predation risk by forming groups. Evolutionary explanations for group formation in previously ungrouped, but loosely associated prey have typically evoked the selfish herd hypothesis. However, despite over 600 studies across a diverse array of taxa, the critical assumptions of this hypothesis have remained collectively untested, owing to several confounding problems in real predator–prey systems. To solve this, we manipulated the domains of danger of Cape fur seal (Arctocephalus pusillus pusillus) decoys to provide evidence that a selfish reduction in a seals'' domain of danger results in a proportional reduction in its predation risk from ambush shark attacks. This behaviour confers a survival advantage to individual seals within a group and explains the evolution of selfish herds in a prey species. These findings empirically elevate Hamilton''s selfish herd hypothesis to more than a ‘theoretical curiosity’.     

Vagrant Subantarctic fur seal at Cape Shirreff,Livingston Island,Antarctica

A vagrant adult male Subantarctic fur seal Arctocephalus tropicalis was observed among Antarctic fur seals A. gazella at Cape Shirreff, Livingston Island, Antarctica, which is located to ~4,190 and ~5,939 km from the nearest breeding colonies of Subantarctic fur seals. Although the colony of origin of this animal and the reason for its movement outside its distribution range are unknown, this sighting shows the high dispersal capacity of this species and provides an insight into possible changes in its distribution. Although this vagrant was not observed with females Antarctic fur seal, news sightings in the future could result in viable hybrid, and introgressive hybridization could represent a threat for Cape Shirreff population recovery, if still the population way to go to recover to presailing levels.     

Characterization of proteins and fatty acid composition in Galapagos fur seal milk. Occurrence of whey and casein protein polymorphisms

1. Milk proteins of the Galapagos fur seal (Arctocephalus galapagoensis) were separated adequately into whey and casein fractions using bovine milk analysis methods. 2. In samples from days 5-30 of lactation 40% of the total proteins were whey and 60% caseins; in mid-lactation, day 150, 25% were whey and 75% casein proteins. 3. Electrophoretic and isoelectric focusing patterns of fur seal whey protein differed widely from bovine patterns, whereas those of caseins were similar. 4. Polymorphisms of fur seal whey and casein proteins were noted and did not seem related to different stages of lactation. 5. C-16 and C-18 fatty acids contributed about 70% of fatty acids; 63% of the total acids in milk fat were unsaturated.