Seaduck engineers in the Arctic Archipelago: nesting eiders deliver marine nutrients and transform the chemistry of island soils, plants, and ponds.

Seabirds are thought to provide ecological services such as the movement of nutrients between marine and terrestrial ecosystems, which may be especially critical to productivity and diversity in nutrient-poor environments. Most Arctic ecosystems are unaffected by local human impacts and are naturally nutrient poor and especially sensitive to warming. Here, we assessed the effects of nesting common eider ducks (Somateria mollissima) on soil, vegetation, and pond sediments on island archipelagoes in Hudson Strait between Nunavut and Québec, Canada. Soil, moss, and pond sediments were significantly higher in nitrogen on islands with large numbers of nesting eiders compared to sites with no nesting birds. The highest concentrations of nitrogen in soils and moss occurred at the margins of ponds on eider islands, which correspond to the areas of highest eider use. δ15N and δ34S values in soils, moss, and sediments indicated substantial marine-derived organic matter inputs at the higher nutrient sites. We propose that by foraging on coastal marine benthic invertebrates and returning to islands to nest, eider ducks bio-transport and concentrate marine-derived nutrients to their colony islands, fertilizing Arctic island ecosystems in the process. As common eiders nest on thousands of low to mid-latitude islands throughout the circumpolar Arctic, these nutrient inputs likely dramatically affect biota and ecosystem functioning throughout the tundra biome.

Foraging behaviour of a continental shelf marine predator, the grey seal (Halichoerus grypus), is associated with in situ, subsurface oceanographic conditions.

BACKGROUND: The heterogeneous oceanographic conditions of continental shelf ecosystems result in a three-dimensionally patchy distribution of prey available to upper-trophic level predators. The association of bio-physical conditions with movement patterns of large marine predators has been demonstrated in diverse taxa. However, obtaining subsurface data that are spatio-temporally relevant to the decisions made by benthically-foraging species can be challenging. METHODS: Between 2009 and 2015, grey seals were captured on Sable Island, Nova Scotia, Canada during summer and fall and instrumented with high-resolution archival GPS tags. These tags recorded location data as well as depth (m), temperature (°C), and light level measurements during dives, until animals returned to the haulout site to breed. Hidden Markov models were used to predict apparent foraging along movement tracks for 79 individuals (59 females, 20 males) every 3 h. In situ measurements were used to estimate chlorophyll-a concentration (mg m- 3) and temperature within the upper-water column (50 m) and temperature and depth at the bottom of dives. As chlorophyll-a could only be estimated from 10:00 to 14:00 AST for dive depths ≥50 m, we formulated two generalized linear mixed-effects models to test the association of predicted grey seal behavioural states with oceanographic conditions and phytoplankton biomass: the first representing conditions of the upper-water column likely to influence primary productivity, and a second model including environmental conditions encountered by grey seals at the bottom of dives, when seals were more likely to be foraging. RESULTS: Predicted grey seal behavioural states were associated with fine-scale chlorophyll-a concentrations and other environmental conditions they encountered across the continental shelf. In the Water Column Model, season had no influence on the probability of observing apparent foraging, but chlorophyll-a, upper-water column temperature, and sex did, with females having a greater probability of foraging than males. In the Bottom Conditions Model, again season had no influence on the probability of apparent foraging, but females were over twice as likely as males to be foraging. CONCLUSIONS: The results of this study highlight the value of in situ measurements of oceanographic properties that can be collected at high temporal resolution by animal-borne data loggers. These data provide insight into how inferred behavioural decisions made by large marine predators, such as the grey seal, may be influenced by fine-scale oceanographic conditions.

Sex-differences in fine-scale home-range use in an upper-trophic level marine predator.

BACKGROUND: The distribution of prey in the ocean is spatially and temporally patchy. How predators respond to this prey patchiness may have consequences on their foraging success, and thus physical condition. The recent ability to record fine-scale movements of marine animals combined with novel home-range analyses that incorporate the dimension of time should permit a better understanding of how individuals utilise different regions of space and the consequences on their foraging success. METHODS: Over a six-year study, we used T-LoCoH (Time-Local Convex Hull) home-range software to model archival GPS (Global Positioning System) data from 81 grey seals to investigate the fine-scale spatio-temporal use of space and the distribution of apparent foraging effort. Regions of home-ranges were classified according to the frequency of return visits (site fidelity) and duration of visits (intensity of use). Generalized linear mixed -effects models were used to test hypotheses on seasonal changes in foraging distribution and behaviour and the role of space-use and state on determining foraging success. RESULTS: Male grey seals had larger home-ranges and core areas than females, and both sexes showed a contraction in home-range and core area in fall leading up to the breeding season compared with summer. Heavier individuals had smaller core areas than lighter ones, suggesting access to higher quality habitat might be limited to those individuals with greater foraging experience and competitive ability. The size of the home-range or core area was not an important predictor of the rate of mass gain. A fine-scale spatio-temporal analysis of habitat use within the home-range provided evidence of intra-annual site fidelity at presumed foraging locations, suggesting predictably in prey distribution. Neither sex nor season were useful predictors for classifying behaviour. Rather, individual identity explained much of the variation in fine-scale behaviour. CONCLUSIONS: Understanding how upper-trophic level marine predators use space provides opportunities to explore the consequences of variation in foraging tactics and their success on fitness. Having knowledge of the drivers that shape this intraspecific variation can contribute toward predicting how these predators may respond to both natural and man-made environmental forcing.

Stable Isotope Models Predict Foraging Habitat of Northern Fur Seals (Callorhinus ursinus) in Alaska.

We developed models to predict foraging habitat of adult female northern fur seals (Callorhinus ursinus) using stable carbon (δ13C) and nitrogen (δ15N) isotope values from plasma and red blood cells. Binomial generalized linear mixed models were developed using blood isotope samples collected from 35 adult female fur seals on three breeding colonies in Alaska during July-October 2006. Satellite location and dive data were used to define habitat use in terms of the proportion of time spent or dives made in different oceanographic/bathymetric domains. For both plasma and red blood cells, the models accurately predicted habitat use for animals that foraged exclusively off or on the continental shelf. The models did not perform as well in predicting habitat use for animals that foraged in both on- and off-shelf habitat; however, sample sizes for these animals were small. Concurrently collected scat, fatty acid, and dive data confirmed that the foraging differences predicted by isotopes were associated with diet differences. Stable isotope samples, dive data, and GPS location data collected from an additional 15 females during August-October 2008 validated the effective use of the models across years. Little within year variation in habitat use was indicated from the comparison between stable isotope values from plasma (representing 1-2 weeks) and red blood cells (representing the prior few months). Constructing predictive models using stable isotopes provides an effective means to assess habitat use at the population level, is inexpensive, and can be applied to other marine predators.

Anatomical patterns of colonization of pets with staphylococcal species in homes of people with methicillin-resistant Staphylococcus aureus (MRSA) skin or soft tissue infection (SSTI).

Methicillin-resistant strains of Staphylococcus aureus (MRSA), Staphylococcus pseudintermedius (MRSP), and other pathogenic staphylococci can cause infections in companion animals and humans. Identification of colonized animals is fundamental to research and practice needs, but harmonized methods have not yet been established. To establish the optimal anatomic site for the recovery of methicillin-resistant coagulase positive staphylococci (CPS), survey data and swabs were collected from 196 pets (dogs, cats, reptiles, birds, fish and pocket pets) that lived in households with an MRSA-infected person. Using broth-enrichment culture and PCR for speciation, S. aureus was identified in 27 of 179 (15%) pets sampled at baseline and 19 of 125 (15%) pets sampled at a three-month follow-up home visit. S. pseudintermedius was isolated from 33 of 179 (18%) pets sampled at baseline and 21 of 125 (17%) of pets sampled at follow-up. The baseline MRSA and MRSP prevalence was 8% and 1% respectively from 145 mammalian pets. The follow-up MRSA and MRSP prevalence was 7% and <1% respectively from 95 mammalian pets. The mouth was the most sensitive single site sampled for isolation of S. aureus and S. pseudintermedius in mammals. In a subset of pets, from which all available isolates were identified, dual carriage of S. aureus and S. pseudintermedius was 22% at baseline and 11% at follow-up. These results identify the mouth as the most sensitive site to screen for pathogenic staphylococci and suggest that it should be included in sampling protocols.

The comparison of manual vs automated disc margin delineation using spectral-domain optical coherence tomography.

AIMS: To examine the impact of manual vs automated disc margin delineation on optic nerve head (ONH) and retinal nerve fiber layer (RNFL) parameters using spectral-domain optical coherence tomography (SDOCT). METHODS: A prospective cohort study consisting of normal, glaucoma suspect (GS) and glaucoma patients who underwent ONH and RNFL measurements using SDOCT technology (RTVue; Optovue Inc.). The retinal pigment epithelium/Bruch's membrane (RPE/BM) complex end points were automatically determined first, and were manually redefined subsequently. Analysis of variance, coefficient of variation (COV), concordance correlation coefficient (CCC), and Bland-Altman plots were used for the analyses. RESULTS: Ninety-nine eyes of 50 subjects (age 68±10 years) consisting of 36 glaucoma, 56 GS, and 7 normal eyes were included. The RNFL thickness measurements were similar (P>0.05) between the two methods of demarcation, except for the inferior-nasal sector (P=0.04). For the ONH measurements, the cup-to-disc (C/D) ratio and rim area showed significant differences between the two methods (P<0.001). COV/CCC values for the ONH parameters were as follows: cup area 17.6%/0.88; cup volume 7.4%/0.91; average C/D ratio 18.1%/0.78; rim area 25.3%/0.69; and rim volume 42.6%/0.71, respectively. CCC/COV values for the RNFL parameters were as follows: average 2.1%/0.98; inferior-temporal quadrant 8.1%/0.79; inferior-nasal quadrant INQ quadrant 12.6%/0.67; SNQ quadrant 7.8%/0.83; and STQ quadrant 7.8%/0.88, respectively. CONCLUSION: An overall high agreement and moderate-substantial concordance was observed between the demarcation methods. Automated disc margin delineation of SDOCT can be used reliably in clinical practice.

Detection of progressive macular thickness loss using optical coherence tomography in glaucoma suspect and glaucomatous eyes.

AIMS: To examine the rate of macular thickness loss using time-domain optical coherence tomography (OCT) in functionally progressing versus non-progressing eyes, determined by standard automated perimetry (SAP). METHODS: Glaucoma suspects (GS) and glaucomatous (G) eyes underwent SAP and OCT imaging every 6 months. Functional progression was determined using pointwise linear regression, defined as 2 contiguous locations losing ≥1.0 dB/year at P<1.0% in the same hemifield. The annual rate of macular thickness loss was calculated from inner and outer regions of the macular map. RESULTS: 72 eyes (43 GS and 29G) with ≥30 months of follow-up were enrolled. Fourteen eyes demonstrated SAP progression. The annual rate of macular thickness loss (µm/year) in progressing eyes was faster (all P<0.05) than non-progressing eyes in temporal outer (-1.90±2.97 vs 0.33±2.77), nasal inner (-1.70±2.66 vs 0.14±2.76), superior inner (-2.15±4.57 vs 0.51±2.99), temporal inner quadrants (-2.58±5.05 vs -0.38±2.34), and the average of inner macular quadrants (-1.84±2.90 vs 0.03±2.10). The rate of loss in the nasal inner (P=0.02) and temporal outer (P=0.02) macular regions was associated with optic disc haemorrhage. CONCLUSIONS: Eyes with SAP progression have significantly greater rates of macular thickness loss consistent with glaucomatous retinal ganglion cell atrophy, as compared with non-progressing eyes.

Repeatability in lactation performance and the consequences for maternal reproductive success in gray seals.

In mammals, the most significant maternal effect on offspring growth during lactation is the ability of females to efficiently transfer milk energy to their neonates. However, despite the importance of the transfer of milk energy to both maternal and offspring fitness, nothing is known about the extent to which variation among females may be attributed to differences in individual quality or environmental variation in natural populations. We measured repeatability over multiple lactation periods in components of lactation performance in free-ranging, multiparous gray seal (Halichoerus grypus) females to examine to what extent variation among females in pup weaning mass may be attributed to inherent differences in their physiological capacity to deliver milk energy. Levels of repeatability were high for milk composition (r = 0.38-0.50), daily milk output (r = 0.46), and the duration of lactation (r = 0.57), demonstrating that there are consistent differences among females in these characters across lactations and that the overall capacity of gray seal females to deliver milk energy to their pups is characteristic of individuals. The repeatability in pup weaning mass (r = 0.48) was consistent with the values for the components of total milk energy output and suggests that, over a large proportion of their reproductive life, individual gray seal females will consistently wean pups with greater or lesser probabilities of survival. Our results suggest that inherent differences among females in their physiological capacity to deliver milk energy may be an important component of variation in individual quality and, thus, lifetime reproductive success in mammals. High levels of repeatability also suggest that components of milk energy transfer may have a significant heritable genetic basis.

Tracing carbon flow in an arctic marine food web using fatty acid-stable isotope analysis.

Global warming and the loss of sea ice threaten to alter patterns of productivity in arctic marine ecosystems because of a likely decline in primary productivity by sea ice algae. Estimates of the contribution of ice algae to total primary production range widely, from just 3 to >50%, and the importance of ice algae to higher trophic levels remains unknown. To help answer this question, we investigated a novel approach to food web studies by combining the two established methods of stable isotope analysis and fatty acid (FA) analysis--we determined the C isotopic composition of individual diatom FA and traced these biomarkers in consumers. Samples were collected near Barrow, Alaska and included ice algae, pelagic phytoplankton, zooplankton, fish, seabirds, pinnipeds and cetaceans. Ice algae and pelagic phytoplankton had distinctive overall FA signatures and clear differences in delta(13)C for two specific diatom FA biomarkers: 16:4n-1 (-24.0+/-2.4 and -30.7+/-0.8 per thousand, respectively) and 20:5n-3 (-18.3+/-2.0 and -26.9+/-0.7 per thousand, respectively). Nearly all delta(13)C values of these two FA in consumers fell between the two stable isotopic end members. A mass balance equation indicated that FA material derived from ice algae, compared to pelagic diatoms, averaged 71% (44-107%) in consumers based on delta(13)C values of 16:4n-1, but only 24% (0-61%) based on 20:5n-3. Our estimates derived from 16:4n-1, which is produced only by diatoms, probably best represented the contribution of ice algae relative to pelagic diatoms. However, many types of algae produce 20:5n-3, so the lower value derived from it likely represented a more realistic estimate of the proportion of ice algae material relative to all other types of phytoplankton. These preliminary results demonstrate the potential value of compound-specific isotope analysis of marine lipids to trace C flow through marine food webs and provide a foundation for future work.

Reproductive performance in grey seals: age-related improvement and senescence in a capital breeder.

1. Three hypotheses have been advanced to account for age-related improvement in performance: the selection hypothesis predicts improved due to the loss of lower quality phenotypes, the constraint hypothesis predicts individuals improve function, and the restraint hypothesis predicts younger individuals forego or reduce effort because of mortality risks. A decline in age-related performance (i.e. senescence) is predicted by mutation accumulation, antagonistic pleiotropy and disposable soma (wear and tear) hypotheses. 2. Using five measures of performance - birth rate, maternal and pup birth mass, pup weaning mass, weaning success and lactation length - we tested these hypotheses concerning age-related change in reproduction in 279 female grey seals (Halichoerus grypus), ages 4-42 years, over a 23-year period between 1983 and 2005 on Sable Island, Nova Scotia. These females produced 2071 pups. 3. Although body mass of primiparous females increased with age (4-7 years) birth mass of their pups did not, but pup weaning mass did. Second- and third-parity females of the same age as primiparous females gave birth to and weaned heavier pups. However, parity and age were dropped from models when maternal body mass was included. 4. The proportion of females giving birth varied significantly with maternal age, increasing in young females and then declining late in life. Weaning success rate also increased rapidly to about 8 years and subsequently declined in females > 32 years. 5. Generalized additive models indicated nonlinear changes in 3 day body mass (i.e. approximately birth mass) and weaning mass of pups as a function of maternal age, after accounting statistically for the effects of maternal body mass. Mixed-effects, repeated-measures models fitted to longitudinal data further supported the conclusion that pup birth mass and weaning mass vary nonlinearly with maternal age and indicated nonlinear changes in lactation duration. 6. We found some support for the constraint hypothesis, but our findings were not consistent with the selection hypothesis or the restraint hypothesis as the basis for improvement in reproductive performance. 7. Senescence was evident in multiple female and offspring traits, indicating the degeneration in function of several physiological systems as predicted by the disposable soma hypothesis.

Weaning mass affects changes in body composition and food intake in harbour seal pups during the first month of independence.

In phocid seals, the transition to nutritional independence is abrupt, with females abandoning their offspring after weaning and returning to sea. We hypothesized that body size at weaning may play an important role in the nature of this transition. We studied the changes in body composition and water flux of newly weaning harbour seals over the first 4-6 wk postweaning. Thirty-three pups were dosed with deuterium oxide to estimate total body water (TBW) and a subset of 24 was dosed twice to estimate changes in body composition and water flux. All pups lost body mass over the study period, but TBW increased during the period of mass loss, indicating continued lean tissue growth. Combined data from this and our early study indicated that heavy (>median mass) pups were relatively fatter (41.0% vs. 37.1%) and had significantly greater total body energy at weaning than did light (< or = median mass) pups. Percentage TBW declined linearly over time in light pups but was constant in heavy pups for the first 19 d postweaning and then declined linearly. Both the temporal pattern and composition of mass loss differed between light and heavy pups. Estimated food intake increased in the second 2 wk of study compared to the first 2 wk, in both light and heavy pups, reflecting increased foraging success but at levels still insufficient to meet daily expenditures of most individuals.

High concentrations of isovaleric acid in the fats of odontocetes: variation and patterns of accumulation in blubber vs. stability in the melon.

Isovaleric acid (iso5:0) is an unusual fatty acid that is important for echolocation and hearing in acoustic tissues of some odontocetes, but its functional significance in blubber is unknown. We examined patterns of accumulation of this compound in blubber in 30 species of odontocetes ( n=299). Iso5:0 concentrations in blubber varied with phylogeny, ontogeny and body topography. Iso5:0 accumulated in greater quantities in superficial/outer blubber than in deep/inner blubber. In the outer blubber of northern right whale and Hector's dolphins, iso5:0 accounted for one-third to one-half of all fatty acids. Total blubber burden of iso5:0 in harbour porpoises represented up to 15 times the amount deposited in the melon. The composition of the melon does not change during starvation in harbour porpoises, supporting the hypothesis that lipids in melon are conserved for a specific function. Some odontocetes continually deposit iso5:0 in blubber after levels in melon have reached asymptotic levels, suggesting independent control of iso5:0 synthesis and storage in these compartments. Dolphins and porpoises inhabiting cold waters possess higher concentrations of iso5:0 in their outer blubber layers than species from warmer regions. We propose that this relationship represents an adaptive secondary role for iso5:0 in maintaining blubber flexibility in cold environments.

Predicting drug-induced agranulocytosis: characterizing neutrophil-generated metabolites of a model compound, DMP 406, and assessing the relevance of an in vitro apoptosis assay for identifying drugs that may cause agranulocytosis.

DMP 406 is a clozapine analogue developed by Dupont-Pharma for the treatment of schizophrenia. Unfortunately it caused agranulocytosis in dogs during preclinical studies. Clozapine also causes agranulocytosis and this is believed to be due to a reactive nitrenium ion metabolite produced by neutrophils. We studied the oxidation of DMP 406 by activated neutrophils and found that the major reactive species that is produced is not a nitrenium ion but rather an imine. This metabolite is similar to the reactive metabolite that has been proposed to be responsible for mianserin-induced agranulocytosis. Therefore we also studied the oxidation of mianserin by activated neutrophils and found that, although the major species is an iminium ion, it also bears a lactam moiety in the piperazine ring resulting from further oxidation. We usually find that HOCl is a good model system for the production of reactive metabolites of drugs that are formed by activated neutrophils, but in the case of both DMP 406 and mianserin, the products produced were significantly different than those formed by activated neutrophils. In contrast, the combination of horseradish peroxidase and hydrogen peroxide (HRP/H(2)O(2)) formed a very similar pattern of products, and this system was used to produce sufficient quantities of metabolites to allow for identification. The reactive metabolites of both DMP 406 and mianserin reacted with a range of nucleophiles, but in many cases the reaction was reversible. The best nucleophile for trapping these reactive metabolites was cyanide. It has been demonstrated that the products of clozapine oxidation by HRP/H(2)O(2), presumably the nitrenium ion, induced apoptosis in neutrophils at therapeutic concentrations of clozapine. It has been suggested that this process is involved in the mechanism of clozapine-induced agranulocytosis. We tested DMP 406 and mianserin in this system to see if the ability of a reactive metabolite of a drug to cause apoptosis could predict the ability of that drug to cause agranulocytosis. We used clozapine as a positive control and we also tested olanzapine, a drug that forms a reactive metabolite similar to that of clozapine but is given at a lower dose and does not cause agranulocytosis. We found that DMP 406 did not increase apoptosis at concentrations below 50 microM, and although mianserin did increase apoptosis at 10 microM this is above the therapeutic concentration. Olanzapine caused an increase in apoptosis at the same concentration as clozapine (1 microM), but because its therapeutic concentration is lower, this concentration was above the pharmacological range. There was no increase in apoptosis with any drug in the absence of HRP/H(2)O(2). These results indicate that this assay is unable to reliably predict the ability of different types of drugs to cause agranulocytosis. This is not a surprising result given that different drugs may induce agranulocytosis by different mechanisms.

Immunotoxicity of explosives-contaminated soil before and after bioremediation.

Soils from the Yorktown Naval Base contaminated with trinitrotoluene (TNT) and other explosives were used to prepare eluates before and after bioremediation using microbial growth amendments in the presence (P1 eluates) or absence (P2 eluates) of exogenous white rot fungus. Effectiveness of bioremediation was examined by several immunotoxicity assays-viability/growth of lymphocytes, cytokine production, and expression of the interleukin-2 (IL-2) receptor-using human peripheral blood mononuclear cells exposed to the eluates. Although TNT concentrations decreased in both P1 and P2 eluates relative to untreated baseline soil (BL) eluates, a recovery in lymphocyte growth/viability and IL-2 secretion was seen with P2 but not P1 eluates relative to BL eluates. IL-2 receptor levels were higher in cells exposed to BL and P2 eluates than when exposed to P1 eluates. Interferon-gamma, tumor necrosis factor-beta, and IL-10 levels were highest in BL and P2 eluates and lowest in P1 eluates. Taken together, these results suggest that treatment of the soil with microbial growth amendments in the absence but not the presence of exogenous white rot fungi lead to partial bioremediation as assessed by lymphocyte functions.

Identification of a reactive metabolite of terbinafine: insights into terbinafine-induced hepatotoxicity.

Oral terbinafine treatment for superficial fungal infections of toe and fingernails is associated with a low incidence (1:45000) of hepatobiliary dysfunction. Due to the rare and unpredictable nature of this adverse drug reaction, the mechanism of toxicity has been hypothesized to be either an uncommon immunological or metabolically mediated effect. However, there is little evidence to support either mechanism, and toxic metabolites of terbinafine have not been identified. We incubated terbinafine with both rat and human liver microsomal protein in the presence of GSH and were able to trap an allylic aldehyde, 7,7-dimethylhept-2-ene-4-ynal (TBF-A), which corresponds to the N-dealkylation product of terbinafine. TBF-A was also prepared synthetically and reacted with excess GSH to yield conjugates with HPLC retention times and mass spectra identical to those generated in the microsomal incubations. The major GSH conjugate, characterized by (1)H NMR, corresponds to addition of GSH in a 1,6-Michael fashion. There remains a second electrophilic site on this metabolite, which can bind either to a second molecule of GSH or to cellular proteins via a 1,4-Michael addition mechanism. Moreover, we demonstrated that the formation of the GSH conjugates was reversible. We speculate that this allylic aldehyde metabolite, formed by liver enzymes and conjugated with GSH, would be transported across the canalicular membrane of hepatocytes and concentrated in the bile. The mono-GSH conjugate, which is still reactive, could bind to hepatobiliary proteins and lead to direct toxicity. Alternatively, it could modify canalicular proteins and lead to an immune-mediated reaction causing cholestatic dysfunction.

Interspecific and intraspecific variation in proximate, mineral, and fatty acid composition of milk in Old World fruit bats (Chiroptera: Pteropodidae).

We examine the effect of body mass on milk composition among Old World fruit bats, including Pteropus pumilus (0.175 kg), Pteropus rodricensus (0.265 kg), Pteropus hypomelanus (0.571 kg), and Pteropus vampyrus (1.133 kg). We describe intra- and interspecific differences in the proximate composition of milk among these four species and the minerals and fatty acids in the milk of the latter two species. There were no differences between species in the concentrations of dry matter, fat, or lactose in milk. However, there were significant, although small, differences in the protein content of milk among species, with protein being significantly greater in P. rodricensus than in P. pumilus and P. hypomelanus and protein being significantly less in P. hypomelanus than in P. rodricensus and P. vampyrus. There were no differences in mineral content between P. hypomelanus and P. vampyrus in milk minerals, but minor differences were evident in fatty acids 12:0, 14:0, 18:0, 18:1n11, and 18:2n6. Our findings suggest that milk composition is relatively constant across lactation for most proximate, mineral, and fatty acid components. We found a significant increase in dry matter and energy across lactation in the concentration of dry matter and energy in P. pumilus and fat in P. hypomelanus. In P. hypomelanus, we found a significant increase in the concentration of fatty acids 10:0 and 20:1n9 and a significant decrease in Iso15 and 20:1n7. No other differences associated with day of lactation were found. These findings suggest that milk composition is generally similar within the genus Pteropus, despite a 6.5-fold difference in body mass between species that we evaluated.

Comparison of the Bligh and Dyer and Folch methods for total lipid determination in a broad range of marine tissue.

For many studies, it is important to measure the total lipid content of biological samples accurately. The Bligh and Dyer method of extraction was developed as a rapid but effective method for determining total lipid content in fish muscle. However, it is also widely used in studies measuring total lipid content of whole fish and other tissues. Although some investigators may have used modified Bligh and Dyer procedures, rarely have modifications been specified nor has their effectiveness been quantitatively evaluated. Thus, we compared this method with that of the classic Folch extraction in determining total lipid content of fish samples ranging from 0.5 to 26.6% lipid. We performed both methods as originally specified, i.e., using the chloroform/methanol/water ratios of 1:2:0.8 and 2:2:1.8 (before and after dilution, respectively) for Bligh and Dyer and of 8:4:3 for Folch, and with the initial solvent/sample ratios of (3+1):1 (Bligh and Dyer) and 20:1 (Folch). We also compared these with several other solvent/sample ratios. In samples containing <2% lipid, the results of the two methods did not differ. However, for samples containing >2% lipid, the Bligh and Dyer method produced significantly lower estimates of lipid content, and this underestimation increased significantly with increasing lipid content of the sample. In the highest lipid samples, lipid content was underestimated by up to 50% using the Bligh and Dyer method. However, we found a highly significant linear relationship between the two methods, which will permit the correction of reported lipid levels in samples previously analyzed using an unmodified Bligh and Dyer extraction. In the future, modifications to procedures and solvent/sample ratios should be described.

Body composition in mink (Mustela vison) kits during 21-42 days postpartum using estimates of hydrogen isotope dilution and direct carcass analysis.

We compared carcass analysis and hydrogen isotope dilution methods to measure total body water (TBW) and body composition in a small altricial carnivore, the mink. Dilution space (D) of mink at 21-42 days of age (n=20), was determined after subcutaneous administration of tritiated water. The same animals were then used to determine TBW and body composition by carcass analysis and to derive predictive empirical relationships between TBW and total body fat, protein and energy. A separate validation set of 27 kits was used to test the accuracy of predicting body composition from TBW. D overestimated TBW by a consistent and predictable 4.1% (R(2)=0.999, P<0.001). Our estimates of fat, protein and energy content, using equations derived from TBW, were not significantly different than those obtained from direct carcass analysis (P>0.980) in either the initial or validation set of mink. TBW was shown to decrease from 81 to 76% and total body protein to increase from 14 to 19% of LBM of the kits from 21 to 42 days of age. Although a rapidly changing hydration state was apparent in neonates, we conclude that when this is taken into account, accurate estimates of body composition can be obtained from hydrogen isotope dilution.

Metabolic compensation during high energy output in fasting, lactating grey seals (Halichoerus grypus): metabolic ceilings revisited.

Lactation is the most energetically expensive period for female mammals and is associated with some of the highest sustained metabolic rates (SusMR) in vertebrates (reported as total energy throughput). Females typically deal with this energy demand by increasing food intake and the structure of the alimentary tract may act as the central constraint to ceilings on SusMR at about seven times resting or standard metabolic rate (SMR). However, demands of lactation may also be met by using a form of metabolic compensation such as reducing locomotor activities or entering torpor. In some phocid seals, cetaceans and bears, females fast throughout lactation and thus cannot offset the high energetic costs of lactation through increased food intake. We demonstrate that fasting grey seal females sustain, for several weeks, one of the highest total daily energy expenditures (DEE; 7.4 x SMR) reported in mammals, while progressively reducing maintenance metabolic expenditures during lactation through means not explained by reduction in lean body mass or behavioural changes. Simultaneously, the energy-exported in milk is progressively increased, associated with increased lipoprotein lipase activity in the mammary gland, resulting in greater offspring growth. Our results suggest that females use compensatory mechanisms to help meet the extraordinary energetic costs of lactation. Additionally, although the concepts of SusMR and ceilings on total DEE may be somewhat different in fasting lactating species, our data on phocid seals demonstrate that metabolic ceilings on milk energy output, in general, are not constrained by the same kind of peripheral limitations as are other energy-consuming tissues. In phocid seals, the high ceilings on DEE during lactation, coupled with metabolic compensation, are undoubtedly important factors enabling shortened lactation.