Wild Grey Seals Research Articles

Vocal learning in animals and humans.

Vocal learning in animals and humans.

Signs of life: Oxygen sensors confirm viability, measure oxygen consumption and provide rapid, effective contamination monitoring for field‐based tissue culture

Abstract Understanding the ecology and evolution of wildlife and domesticated species requires knowledge of their physiological responses to environmental change and the constraints under which they operate. However, whole animal experiments are often limited in sample size and can be logistically and ethically challenging. Culture techniques represent a powerful approach, but are used infrequently in field research due to practical constraints. We used minimal tissue culture equipment in a remote field site for in vitro explant experiments using blubber from wild grey seals Halichoerus grypus. Assessing explant viability and detecting microbial contamination in remote field sites, where facilities are often small, unspecialised and more vulnerable to bacterial or fungal infection, present major challenges. We investigated whether oxygen‐sensitive planar optodes (OSPO) in closed system respirometry could be used to assess oxygen consumption by blubber explants from suckling and fasting wild seal pups as a proxy for viability. We also explored whether OSPOs could provide rapid information on whole animal relevant physiological metrics by determining whether explant oxygen consumption correlated with the nutritional state of the animal, blubber depth and other tissue metabolic properties, including glucose uptake, lactate production and lipolysis. Vials containing blubber explants consumed significant amounts of oxygen compared to controls, showing tissues were metabolically active. Oxygen consumption differed between nutritional states and blubber tissue depth. These differences were reflected in other tissue metabolic properties. Dissolved oxygen levels remained consistent over 24 hr in 94% of control vials containing only culture media. In 6% of control vials extremely rapid oxygen consumption preceded, by 2–3 days, colour changes in the phenol‐red containing media that indicate lactic acidosis from microbial metabolic activity. Oxygen use in control vials was, therefore, an effective monitoring system that provided vital early warning of media contamination, allowing stocks to be discarded, which prevented erroneous results and avoided waste of valuable field time and irreplaceable samples. OSPO are thus a useful tool for simultaneously assessing tissue oxygen consumption, investigating functional physiological differences and monitoring microbiological contamination in culture experiments, particularly in field laboratories studying live tissues from wildlife.

High oxytocin infants gain more mass with no additional maternal energetic costs in wild grey seals (Halichoerus grypus)

Maximising infant survival requires secure attachments and appropriate behaviours between parents and offspring. Oxytocin is vital for parent-offspring bonding and behaviour. It also modulates energetic balance and neural pathways regulating feeding. However, to date the connections between these two areas of the hormone’s functionality are poorly defined. We demonstrate that grey seal (Halichoerus grypus) mothers with high oxytocin levels produce pups with high oxytocin levels throughout lactation, and show for the first time a link between endogenous infant oxytocin levels and rates of mass gain prior to weaning. High oxytocin infants gained mass at a greater rate without additional energetic cost to their mothers. Increased mass gain in infants was not due to increased nursing, and there was no link between maternal mass loss rates and plasma oxytocin concentrations. Increased mass gain rates within high oxytocin infants may be due to changes in individual behaviour and energy expenditure or oxytocin impacting on tissue formation. Infancy is a crucial time for growth and development, and our findings connect the oxytocin driven mechanisms for parent-infant bonding with the energetics underlying parental care. Our study demonstrates that oxytocin release may connect optimal parental or social environments with direct physiological advantages for individual development.

BRUCELLA PINNIPEDIALIS IN GREY SEALS ( HALICHOERUS GRYPUS) AND HARBOR SEALS ( PHOCA VITULINA) IN THE NETHERLANDS.

Brucellosis is a zoonotic disease with terrestrial or marine wildlife animals as potential reservoirs for the disease in livestock and human populations. The primary aim of this study was to assess the presence of Brucella pinnipedialis in marine mammals living along the Dutch coast and to observe a possible correlation between the presence of B. pinnipedialis and accompanying pathology found in infected animals. The overall prevalence of Brucella spp. antibodies in sera from healthy wild grey seals ( Halichoerus grypus; n=11) and harbor seals ( Phoca vitulina; n=40), collected between 2007 and 2013 ranged from 25% to 43%. Additionally, tissue samples of harbor seals collected along the Dutch shores between 2009 and 2012, were tested for the presence of Brucella spp. In total, 77% (30/39) seals were found to be positive for Brucella by IS 711 real-time PCR in one or more tissue samples, including pulmonary nematodes. Viable Brucella was cultured from 40% (12/30) real-time PCR-positive seals, and was isolated from liver, lung, pulmonary lymph node, pulmonary nematode, or spleen, but not from any PCR-negative seals. Tissue samples from lung and pulmonary lymph nodes were the main source of viable Brucella bacteria. All isolates were typed as B. pinnipedialis by multiple-locus variable number of tandem repeats analysis-16 clustering and matrix-assisted laser desorption ionization-time of flight mass spectrometry, and of sequence type ST25 by multilocus sequence typing analysis. No correlation was observed between Brucella infection and pathology. This report displays the isolation and identification of B. pinnipedialis in marine mammals in the Dutch part of the Atlantic Ocean.

Using blubber explants to investigate adipose function in grey seals: glycolytic, lipolytic and gene expression responses to glucose and hydrocortisone

Adipose tissue is fundamental to energy balance, which underpins fitness and survival. Knowledge of adipose regulation in animals that undergo rapid fat deposition and mobilisation aids understanding of their energetic responses to rapid environmental change. Tissue explants can be used to investigate adipose regulation in wildlife species with large fat reserves, when opportunities for organismal experimental work are limited. We investigated glucose removal, lactate, glycerol and NEFA accumulation in media, and metabolic gene expression in blubber explants from wild grey seals. Glycolysis was higher in explants incubated in 25 mM glucose (HG) for 24 h compared to controls (C: 5.5 mM glucose). Adipose-derived lactate likely contributes to high endogenous glucose production in seals. Lipolysis was not stimulated by HG or high hydrocortisone (HC: 500 nM hydrocortisone) and was lower in heavier animals. HC caused NEFA accumulation in media to decrease by ~30% relative to C in females, indicative of increased lipogenesis. Lipolysis was higher in males than females in C and HG conditions. Lower relative abundance of 11-β-hydroxysteroid dehydrogenase 1 mRNA in HG explants suggests glucose involvement in blubber cortisol sensitivity. Our findings can help predict energy balance responses to stress and nutritional state in seals, and highlight the use of explants to study fat tissue function in wildlife.

Positive social behaviours are induced and retained after oxytocin manipulations mimicking endogenous concentrations in a wild mammal.

The neuropeptide hormone oxytocin modulates numerous social and parental behaviours across a wide range of species, including humans. We conducted manipulation experiments on wild grey seals (Halichoerus grypus) to determine whether oxytocin increases proximity-seeking behaviour, which has previously been correlated with endogenous oxytocin concentrations in wild seal populations. Pairs of seals that had never met previously were given intravenous injections of 0.41 µg kg−1 oxytocin or saline and were observed for 1 h post-manipulation. The dose was designed to mimic endogenous oxytocin concentrations during the observation period, and is one of the lowest doses used to manipulate behaviour to date. Seals given oxytocin spent significantly more time in close proximity to each other, confirming that oxytocin causes conspecifics to seek others out and remain close to one another. Aggressive and investigative behaviours also significantly fell after oxytocin manipulations. Despite using a minimal oxytocin dose, pro-social behavioural changes unexpectedly persisted for 2 days despite rapid dose clearance from circulation post-injection. This study verifies that oxytocin promotes individuals staying together, demonstrating how the hormone can form positive feedback loops of oxytocin release following conspecific stimuli, increased motivation to remain in close proximity and additional oxytocin release from stimuli received while in close proximity.

Obtaining accurate glucose measurements from wild animals under field conditions: comparing a hand held glucometer with a standard laboratory technique in grey seals.

Glucose is an important metabolic fuel and circulating levels are tightly regulated in most mammals, but can drop when body fuel reserves become critically low. Glucose is mobilized rapidly from liver and muscle during stress in response to increased circulating cortisol. Blood glucose levels can thus be of value in conservation as an indicator of nutritional status and may be a useful, rapid assessment marker for acute or chronic stress. However, seals show unusual glucose regulation: circulating levels are high and insulin sensitivity is limited. Accurate blood glucose measurement is therefore vital to enable meaningful health and physiological assessments in captive, wild or rehabilitated seals and to explore its utility as a marker of conservation relevance in these animals. Point-of-care devices are simple, portable, relatively cheap and use less blood compared with traditional sampling approaches, making them useful in conservation-related monitoring. We investigated the accuracy of a hand-held glucometer for 'instant' field measurement of blood glucose, compared with blood drawing followed by laboratory testing, in wild grey seals (Halichoerus grypus), a species used as an indicator for Good Environmental Status in European waters. The glucometer showed high precision, but low accuracy, relative to laboratory measurements, and was least accurate at extreme values. It did not provide a reliable alternative to plasma analysis. Poor correlation between methods may be due to suboptimal field conditions, greater and more variable haematocrit, faster erythrocyte settling rate and/or lipaemia in seals. Glucometers must therefore be rigorously tested before use in new species and demographic groups. Sampling, processing and glucose determination methods have major implications for conclusions regarding glucose regulation, and health assessment in seals generally, which is important in species of conservation concern and in development of circulating glucose as a marker of stress or nutritional state for use in management and monitoring.

Play Behavior of Wild Grey Seals (Halichoerus grypus): Effects of Haulout Group Size and Composition

Play Behavior of Wild Grey Seals (Halichoerus grypus): Effects of Haulout Group Size and Composition

Maternal Oxytocin Is Linked to Close Mother-Infant Proximity in Grey Seals (Halichoerus grypus).

Maternal behaviour is a crucial component of reproduction in all mammals; however the quality of care that mothers give to infants can vary greatly. It is vital to document variation in maternal behaviour caused by the physiological processes controlling its expression. This underlying physiology should be conserved throughout reproductive events and should be replicated across all individuals of a species; therefore, any correlates to maternal care quality may be present across many individuals or contexts. Oxytocin modulates the initiation and expression of maternal behaviour in mammals; therefore we tested whether maternal plasma oxytocin concentrations correlated to key maternal behaviours in wild grey seals (Halichoerus grypus). Plasma oxytocin concentrations in non-breeding individuals (4.3 ±0.5 pg/ml) were significantly lower than those in mothers with dependent pups in both early (8.2 ±0.8 pg/ml) and late (6.9 ±0.7 pg/ml) lactation. Maternal plasma oxytocin concentrations were not correlated to the amount of nursing prior to sampling, or a mother’s nursing intensity throughout the dependant period. Mothers with high plasma oxytocin concentrations stayed closer to their pups, reducing the likelihood of mother-pup separation during lactation which is credited with causing starvation, the largest cause of pup mortality in grey seals. This is the first study to link endogenous oxytocin concentrations in wild mammalian mothers with any type of maternal behaviour. Oxytocin’s structure and function is widely conserved across mammalian mothers, including humans. Defining the impact the oxytocin system has on maternal behaviour highlights relationships that may occur across many individuals or species, and such behaviours heavily influence infant development and an individual’s lifetime reproductive success.

Naso-pharyngeal mites Halarachne halichoeri (Allman, 1847) in Grey seals stranded on the NW Spanish Atlantic Coast

In North Atlantic European waters, the nasal mite Halarachne halichoeri has been described affecting Grey seals (Halichoerus grypus) producing different levels of respiratory disease. This study provides data on the prevalence, clinical signs and produced macro-pathology of this parasite mite infecting juvenile wild Grey seals stranded in North-Western Spanish coast. Among the 25 seals examined during the study, a total of 19 had nasal mites in their respiratory upper ways, including adult and larval stages. This represented a percentage of prevalence of 76±8.37. All the live positive seals presented a typical clinical symptomatology associated to upper respiratory tract infections. In dead positive seals, a light to intense sinusitis could be diagnosed macroscopically. The presence of the parasite in the nasal sinuses appears as the primary cause of the high respiratory tract symptomatology presented in most of the juvenile seals stranded in the north coast of Spain. Ultrastructural characterization by scanning electronic microscopy (SEM) has confirmed the taxonomic status of the mite. This identification of H. halichoeri represents the first description of the occurrence of this parasite in Southern Europe.

Estimating field metabolic rates of pinnipeds: doubly labelled water gets the seal of approval

Summary 1. Measures of the field metabolic rate of marine mammals are extremely difficult to make but they are essential for assessing the impacts of marine mammals on prey populations, and for assessing dive performance in relation to aerobic limits. 2. The doubly labelled water (DLW) method is an isotope-based technique for the estimation of the CO 2 production, and hence energy expenditure, of free-living animals. Estimates of field metabolic rate (FMR) from DLW in pinnipeds to date are extremely high and at the upper range for most mammals. DLW has previously been validated in pinnipeds but logistical difficulties meant for these validations were less than ideal, and it has been hypothesised that DLW may overestimate FMR in these animals. 3. To test this hypothesis, we used DLW and simultaneous open-flow respirometry to determine the daily energy expenditures (DEE) of wild grey seals ( Halichoerus grypus ) held temporarily in a captive facility, during 4‐5 days of simulated foraging at sea. Comparing DEE from DLW and respirometry, we found that DLW predicted DEE accurately (average difference between the two estimates was 0·7% SD = 17% n = 31), but as with validations of other species there was a large range of individual errors (from ‐39% to +44%). 4. The results dispel the doubts surrounding the use of DLW as a field method for estimating energy expenditure in grey seals, and by implication other pinnipeds, and simultaneously open a series of questions about their ability to maintain surprisingly high metabolic rates for protracted periods. 5. We make a number of recommendations for future studies of pinniped FMR using DLW. We suggest use of the Speakman two-pool calculation will be most appropriate. Studies should aim for enrichment levels as high as economically feasible but to at least 150 p.p.m. above background for the O 2 isotope. Measurement periods should be extended between one and two half-lives (5‐10 days for a typical foraging seal). We also encourage the calculation and presentation of estimates of precision in estimates of FMR.

Seasonal variation in the metabolic rate and body composition of female grey seals: fat conservation prior to high-cost reproduction in a capital breeder?

Many animals rely on stored energy through periods of high energy demand or low energy availability or both. A variety of mechanisms may be employed to attain and conserve energy for such periods. Wild grey seals demonstrate seasonal patterns of energy storage and foraging behaviour that appear to maximize the allocation of energy to reproduction--a period characterized by both high energy demand and low food availability. We examined seasonal patterns in resting rates of oxygen consumption as a proxy for metabolic rate (RMR) and body composition in female grey seals (four adults and six juveniles), testing the hypothesis that adults would show seasonal changes in RMR related to the reproductive cycle but that juveniles would not. There was significant seasonal variation in rates of resting oxygen consumption of adult females, with rates being highest in the spring and declining through the summer months into autumn. This variation was not related to changes in water temperature. Adults increased in total body mass and in fat content during the same spring to autumn period that RMR declined. RMR of juveniles showed no clear seasonal patterns, but did increase with increasing mass. These data support the hypothesis that seasonal variation in RMR in female grey seals is related to the high costs of breeding.

Evidence for a long-lasting single administration contraceptive vaccine in wild grey seals

A single-administration birth control vaccine based on liposome delivery of porcine zona pellucida antigens reduced pup production in grey seals ( Halichoerus grypus) by about 90%. Anti-porcine zona pellucida titers of individual seals with two or more recaptures were variable but without a diminishing trend during the 5 year post-immunization period. Seals that produced at least one or more pups during the 2–5 year post-immunization period when the vaccine is fully effective, had an average anti-porcine zona pellucida titer of 5% of the reference serum. In contrast, the subset of seals that did not reproduce but were recaptured during the breeding season had an average titer of 31% of the reference serum. As measured by antibody titers and pup production, there were no differences in efficacy of the vaccine in 14-, 20- and 21-year-old female grey seals.

Detailed observations of suckling behaviour of a Grey seal Halichoerus grypus at Harderwijk Marine Mammal Park

In each of the five years, 1987 to 1991, a pair of Grey seals Halichoerus grypus produced a pup. For each birth the mother and pup were kept under constant observation and a variety of suckling parameters were recorded. In January 1991 the pup was born in an indoor suckling enclosure where in 1987 and 1990 other infants had been reared. Most suckling sessions were initiated by the pup vocalizing followed by the mother presenting her nipples. The pup ended 89% of the sessions. Throughout the 17‐day suckling period, total daily suckling time, suckling session length and number of pauses in suckling sessions all increased. The length of non‐suckling intervals decreased. Even in the absence of vocalizations the mother appeared to be able to judge when the pup wanted to suckle, probably alerted by the waking‐up behaviour of the infant. The ♀ flippered before 43%, during 41% and after 76% of all suckling sessions. The pup flippered before 12%, during 13% and after 17%) of the suckling sessions. The mother's food intake was low for a few days immediately before and for three days after the birth. After this her food intake increased to 5 kg per day until day 9, then decreased to 0–5 kg one day before weaning. The ♀ ate more after weaning and copulation than during the suckling period when she lost a mean 3 kg per day. The pup gained a mean 2 kg per day and by day 43 he began to catch live fish. When the pup started to eat thawed marine fish the colour of his mystacial vibrissae started to change and about two weeks later they went from black to transparent. At the age of 120 days the pup was released into the North Sea to join a group of wild Grey seals.

The Metabolic Characteristics of the Locomotory Muscles of Grey Seals (Halichoerus Grypus), Harbour Seals (Phoca Vitulina) and Antarctic Fur Seals (Arctocephalus Gazella)

It is not known precisely how marine mammals are able to maintain muscle function during active swimming in breath-hold dives, when ventilation stops and heart rate falls. Examination of muscle biochemistry and histochemistry can provide information on the relative importance of different metabolic pathways, the contractile potential of the muscle fibres, the oxygen storage capacity of the muscle and the capillary distribution in these animals. In this study, samples of locomotory muscle were taken from wild grey seals (Halichoerus grypus), harbour seals (Phoca vitulina) and Antarctic fur seals (Arctocephalus gazella); Wistar rat muscle was analysed for comparative purposes. Activities of citrate synthase and beta-hydroxyacyl CoA dehydrogenase were higher in the harbour seal muscle than in the grey seal muscle, suggesting that harbour seals have a greater aerobic capacity. Both phocid muscles had a greater reliance on fatty acid oxidation than the fur seal or rat muscles. The myoglobin data demonstrate that the grey seals have the highest oxygen storage capacity of the three pinniped species, which correlates with their greater diving ability. Myoglobin levels were higher in all three pinniped species than in the Wistar rat. The fibre type compositions suggest that the muscles from the fur seals have higher glycolytic capacities than those of the phocid seals [fur seal pectoralis, 7% slow-twitch oxidative fibres (SO), 25% fast-twitch oxidative glycolytic fibres (FOG), 68% fast-twitch glycolytic fibres (FG); grey seal 57% SO, 5% FOG, 38% FG; area per cents]. However, the pectoralis muscle of the fur seal, although the most glycolytic of the pinniped muscles studied, has the highest capillary density, which indicates a high capacity for fuel distribution. These results show that, while pinniped muscle has an increased oxygen storage potential compared with the muscle of a typical terrestrial mammal, there are no distinct adaptations for diving in the enzyme pathways or fibre type distributions of the pinniped muscle. However, the muscle characteristics of each species can be related to its diving behaviour and foraging strategy.

Gas Exchange of Captive Freely Diving Grey Seals (Halichoerus Grypus)

When at sea, phocids dive for long periods and spend a high percentage of their time submerged. This behaviour requires some combination of an increased oxygen storage capacity, rapid oxygen loading at the surface and reduced oxygen utilisation when submerged. To assess these adaptations, breath-by-breath ventilation was studied in four adult grey seals (two male, two female, 160-250 kg), freely diving in a large outdoor tank where surface access was restricted to one breathing hole. The dive patterns obtained were similar to those recorded from freely diving wild grey seals. Respiratory frequency during the surface periods was 40% higher than that estimated from allometric relationships (19.4 +/- 0.7 breaths min-1), and tidal volume (6.3 +/- 1.21) was approximately five times higher than that estimated from allometric relationships. These adaptations produce a high minute volume and enable gas exchange to occur at the surface. Mean oxygen consumption rate (VO2, measured for a dive+surface cycle) decreased with increasing dive duration. The aerobic dive limit was estimated as 9.6 min for a 150 kg grey seal (using the overall average VO2 of 5.2 ml O2 min-1 kg-1), which is consistent with results from freely diving wild grey seals (only 6% of dives exceeded 10 min). End-tidal oxygen values varied during a surface period, following a U-shaped curve, which suggests that there is limited oxygen uptake from the lung and/or blood oxygen stores during dives. This result was unexpected and indicates that these seals are utilising substantial physiological responses to conserve oxygen, even during shallow voluntary diving.