Diet estimation based on an integrated mixed prey feeding experiment using Arctocephalus seals

Food web models depend on identifying which taxa are eaten and in what proportion they are consumed. Arctocephalus seals are generalist foragers and are an ongoing focus of Southern Hemisphere marine ecosystem research. This is the first feeding experiment to use Arctocephalus spp. to assess the utility of hard part scat analysis for diet estimation, based on mixed prey diets integrated over several days. Recovery rates of otoliths were extremely low for all taxa (0-9%). Although we could not collect scats produced during a 90 min period each day, during which the seals had access to a large pool, this result could not be attributed to otolith robustness, pinniped species or class, activity level, meal size or frequency, or fat content of the diet. We conclude that the unusually low recovery rates in this study may be due to unaccounted scats produced during 90 min of each day, if they contained otolith numbers an order of magnitude greater than all otoliths retrieved from scats produced during the other 22.5 h of each day, and/or may be related to the digestive processing of a mixed prey diet. Our study demonstrates the inadequacy of using otoliths in field collected scats for diet estimation due to the high level of unexplained variability of otolith occurrence in scats. We also identify two new potential sources of this variability. These are variability in numbers of otoliths per scat depending on activity level when a scat is excreted, and variability in recovery rates of otoliths as a function of the complexity of the diet.     

INVESTIGATING THE BIASES IN THE USE OF HARD PREY REMAINS TO IDENTIFY DIET COMPOSITION USING ANTARCTIC FUR SEALS (ARCTOCEPHALUS GAZELLA) IN CAPTIVE FEEDING TRIALS

The analysis of pinniped scats has been used to quantify their diet, using prey remains to identify species and to estimate the numbers and sizes of prey consumed. There are, however, potential biases involved with scat analysis and, for this method to be used successfully, these biases need to be quantified. Thirty-six Antarctic fur seals ( Arctocephalus gazella ) were fed meals of exclusively either fish, squid, or krill and their scats were collected and analyzed. The major sources of error in the analysis of prey remains from scats were the differential erosion and passage rate of items in relation to their size. However, using simple correction functions, such as those which model otolith erosion, it is possible to reduce these errors. Using plastic beads as dietary markers showed recovery rates were negatively related to their size. Larger squid beaks had lower recovery rates than smaller beaks, but there was no size-related bias in the recovery rates of krill carapaces. Only 33% of the squid beaks and 27% of the otoliths originally fed were recovered from the scats. Recovery rates were greater for squid (77%) and fish (50%) eye lenses and these structures gave a better estimate of numbers eaten. Differences found between experimentally derived and published regression equations (used to calculate prey sizes eaten from prey remains) highlights the need for regression equations based on local prey characteristics, if these are to be used with any success to describe the prey eaten.     

Assessment of errors associated with harbour seal (Phoca vitulina) faecal sampling

Six harbour seals, ages 4–8 years, were held as pairs in a 10 times 20 times 2 m tank filled with sea water, and on 60 occasions were fed a meal of a specific species of fish or cephalopod of known size. The tank was drained periodically, and harbour seal faeces were collected on a 0.5 mm sieve. Number and size of otoliths and beaks found in faeces were determined. Fifty-eight percent of 670 fish and 37% of 36 cephalopods fed to harbour seals were represented by their otoliths or beaks in faeces. Estimated number of prey consumed was determined from the greatest number of left or right otoliths or upper or lower beaks collected in faeces. Estimated length ofprey was determined from measurements of otoliths and beaks recovered in the tank and relationships of otolith and beak measurements to prey length. Estimated number of fish eaten was not significantly different among pairs of harbour seals, but was different among species of fishes. Only 24–35% of fish species with small otoliths were represented in faeces, whereas more robust otoliths from other species were less apt to be completely dissolved. Estimated length of fishes was significantly less than lengths of fishes fed to harbour seals in 39 (76.5%) of 51 trials. Cephalopod beaks were not affected by passage through the harbour seal digestive tract. Amount of otolith dissolution was not related to species of fish; estimated fish length was underestimated by an average 27.5%. Although some (7.4%) of the otoliths were collected within 100 h after the fish were ingested, more than 90% were recovered within 24 h after the fish was eaten. Correction factors were developed which will allow researchers to estimate more reliably number and size of fish and cephalopod prey eaten by harbour seals.     

A captive feeding study with the Pacific harbor seal (Phoca vitulina richardii): Implications for scat analysis

Seven prey species ( n _total > 2,700) were fed to seven captive male Pacific harbor seals ( Phoca vitulina richardii ) in 177 experimental meals to quantify biases associated with scat analysis and current consumption models. Hard parts from an individual meal were recovered in an average of 3.8 ± 1.8 scats (range 1–10; mean ± SD). Overall, 57.7 ± 33.2% of otoliths and 89.5 ± 15.5% of squid beaks were recovered. Recovery rates varied, and prey with smaller, fragile otoliths were recovered in lesser quantities than prey with larger, robust otoliths. Recovery rates of all prey except pink salmon were improved by a mean of 31.7% when all diagnostic structures were included in estimates. Estimated recovery of pink salmon was 9.5 times that fed seals based on the all-structure technique. Mean length reduction of recovered otoliths was 20.4 ± 10.1%. Correction factors calculated from average length reduction improved length estimates for all fish species. Grade-specific length correction factors (gLCFs) reduced variability in all of the estimates and significantly improved estimates of prey with highly eroded otoliths including Pacific hake and shortbelly rockfish. The Biomass Reconstruction (BR) model accurately predicted biomass consumption within 4% of known consumption, whereas estimates based on frequency of occurrence were inaccurate.     

FEEDING METHOD AFFECTS OTOLITH DIGESTION IN CAPTIVE GRAY SEALS: IMPLICATIONS FOR DIET COMPOSITION ESTIMATION

Analysis of prey structures from scats has become widely used for assessing the diet of free-ranging seals. Captive feeding experiments have played an important role in the development of scat analysis but methodology has varied. We investigated whether using an experimental otolith-carrier species or in situ otolith experimental prey to feed haddock, plaice, and sand eel otoliths to two captive gray seals affected the amount of otolith digestion, and therefore our ability to estimate fish size and diet composition. Otolith recovery rates varied among the three prey species, but were not affected by feeding method. Otoliths were more digested in carrier experiments; resulting digestion coefficients were greater than those from in situ experiments. As a result, while the lengths of whole fish fed experimentally could be accurately predicted by applying in situ digestion coefficients, the lengths of these fish could not be accurately predicted using carrier digestion coefficients. Consequently, the estimated proportion by weight of each species in the diet can change markedly when carrier, as opposed to in situ , digestion coefficients are used. The implications for diet composition estimation are significant.     

Large‐scale molecular barcoding of prey DNA reveals predictors of intrapopulation feeding diversity in a marine predator

Predator–prey interactions are critical in understanding how communities function. However, we need to describe intraspecific variation in diet to accurately depict those interactions. Harbor seals (Phoca vitulina) are an abundant marine predator that prey on species of conservation concern. We estimated intrapopulation feeding diversity (variation in feeding habits between individuals of the same species) of harbor seals in the Salish Sea. Estimates of feeding diversity were examined relative to sex, month, and location using a novel approach that combined molecular techniques, repeated cross‐sectional sampling of scat, and a specialization metric (within‐individual consistency in diet measured by the Proportional Similarity Index ()). Based on 1,083 scat samples collected from five haul‐out sites during four nonsequential years, we quantified diet using metabarcoding techniques and determined the sex of the scat depositor using a molecular assay. Results suggest that intrapopulation feeding diversity was present. Specialization was high over short periods (24–48 hr,  = 0.392, 95% CI = 0.013, R = 100,000) and variable in time and space. Females showed more specialization than males, particularly during summer and fall. Additionally, demersal and benthic prey species were correlated with more specialized diets. The latter finding suggests that this type of prey likely requires specific foraging strategies and that there are trade‐offs between pelagic and benthic foraging styles for harbor seals. This differential feeding on prey species, as well as between sexes of harbor seals, indicates that predator–prey interactions in harbor seals are complex and that each sex may have a different impact on species of conservation concern. As such, describing intrapopulation feeding diversity may unravel hitherto unknown complex predator–prey interactions in the community.     

EFFECTS OF MEAL SIZE ON OTOLITH RECOVERY FROM FECAL SAMPLES OF GRAY AND HARBOR SEAL PUPS

Recovered otoliths from pinniped feces provide valuable information on diet composition and prey size. We studied the effect of meal size on otolith recovery from the feces of one harbor and eight gray seal pups. Each of 11 experiments comprised a half-ration meal, a period of fecal collection, a 1.5-or double-ration meal again followed by a period of fecal collection. A significantly lower percentage of Atlantic herring otoliths were recovered from half-ration meals (25%± 12.5% in the harbor seal, 8.6%± 6.9% in eight gray seals) than from 1.5- or double-ration meals (62.5%± 3.1 % in the harbor seal, 32.8%± 23.5% in gray seals). Meal size also significantly affected the percentage of Atlantic cod otoliths recovered from gray seal feces (65.0%± 26.3% from half ration, 98.3%± 2.9% from 1.5 ration). For both size meals, recovered cod otoliths were more significantly eroded than herring otoliths. The development of correction factors to account for the effects of digestion will need to consider the distribution of meal sizes of free-ranging pinnipeds.     

Fish diet of the Antarctic fur seal,Arctocephalus gazella,in summer,at Stranger Point,King George Island,South Shetland Islands

 In February 1992, 34 faecal samples from non-breeding male Antarctic fur seals, Arctocephalus gazella, were collected at Stranger Point, King George Island, South Shetlands. Fish constituted an important part of the diet, occurring in 90% of those scats containing prey remains. From 1162 otoliths found in the faeces, 1086 were identified to at least family level. Myctophids and nototheniids represented together almost 90% of the fish eaten. The dominant species were Gymnoscopelus nicholsi, Pleuragramma antarcticum and Electrona antarctica, contributing 33.3%, 30.8% and 12.0% of the otoliths respectively. The standard length of these three species was estimated from otoliths with little or no signs of erosion. This study showed that fur seals fed mainly on pelagic fish species that are often associated with krill. These findings are corroborated by fur seal diving patterns. Received: 7 October 1994/Accepted: 5 October 1995     

Quantifying sequence proportions in a DNA‐based diet study using Ion Torrent amplicon sequencing: which counts count?

A goal of many environmental DNA barcoding studies is to infer quantitative information about relative abundances of different taxa based on sequence read proportions generated by high‐throughput sequencing. However, potential biases associated with this approach are only beginning to be examined. We sequenced DNA amplified from faeces (scats) of captive harbour seals (Phoca vitulina) to investigate whether sequence counts could be used to quantify the seals’ diet. Seals were fed fish in fixed proportions, a chordate‐specific mitochondrial 16S marker was amplified from scat DNA and amplicons sequenced using an Ion Torrent PGM?. For a given set of bioinformatic parameters, there was generally low variability between scat samples in proportions of prey species sequences recovered. However, proportions varied substantially depending on sequencing direction, level of quality filtering (due to differences in sequence quality between species) and minimum read length considered. Short primer tags used to identify individual samples also influenced species proportions. In addition, there were complex interactions between factors; for example, the effect of quality filtering was influenced by the primer tag and sequencing direction. Resequencing of a subset of samples revealed some, but not all, biases were consistent between runs. Less stringent data filtering (based on quality scores or read length) generally produced more consistent proportional data, but overall proportions of sequences were very different than dietary mass proportions, indicating additional technical or biological biases are present. Our findings highlight that quantitative interpretations of sequence proportions generated via high‐throughput sequencing will require careful experimental design and thoughtful data analysis.     

Investigating a simplified method for noninvasive genetic sampling in East African mammals using silica dried scat swabs

Swabbing scat has proved to be an effective noninvasive method to collect DNA from mammals in the field. Previously, this method has relied on preservative liquids or freezing to preserve the DNA collected on swabs. In this study, we determine the effectiveness of using silica to simply dry the swab in field as an alternative way to prevent DNA degredation. Four species were included in the study; reticulated giraffe, impala, fringe‐eared oryx, and lion. Swabs were taken at multiple time points for giraffe and impala scat samples, with the lion and oryx sampled opportunistically. Mitochondrial DNA was successfully amplified and sequenced from scat swabs from all species; however, effectiveness varied between species, with 81.8% amplification success rate from swabs taken from impala scat compared to 25% amplification success rate in giraffe. This variation in success rate was overcome by taking multiple swabs, thus increasing the probability of a successful amplification. The true merit of this method is in its simplicity and cheapness; no preservative liquids were required to be brought into the field, at no stage in the 2 weeks of field sampling were samples frozen, and no commercial kits were used for DNA extraction.     

THE DISTRIBUTION, ABUNDANCE AND SELECTED PREY OF THE HARBOR SEAL, PHOCA VITULINA CONCOLOR, IN SOUTHERN NEW ENGLAND

The seasonal distribution and abundance of harbor seals occurring south of Maine were documented by counting the number of seals at traditional haulout locations. The average number of seals counted during each survey in Massachusetts and New Hampshire was 3,560 ± 255 (95% CI), 1983–1987. The maximum number of seals counted on any individual survey was 4,736 individuals. Fifty percent of all the surveys since January 1985 have resulted in counts greater than 4,000 seals reflecting a 27% increase in the abundance of seals in our study area since that date. Seventy-five percent of the seals in southern New England are located at haulout sites on Cape Cod and Nantucket Island. The largest aggregation of seals in the eastern United States occurs mid-winter at Monomoy Island and adjacent shoals. A single high count of 1,672 seals occurred at this site during the study period. An additional 271–374 seals were also counted in Rhode Island, Connecticut and eastern Long Island Sound during surveys conducted in March 1986 and 1987. The American sandlance Ammodytes americanus was the single dominant prey item of harbor seals in waters adjacent to Cape Cod based on the modified frequency of occurrence of each prey species in scat samples collected from three haulout sites on Cape Cod between 1984–1987. During January and February sandlance was the near exclusive prey item at Monomoy (99%, n= 80). During March and April, the frequency of Atlantic herring Clupea harengus increased in the scat samples at this site. Regional differences in the diet of seals reflect distinct prey communities throughout the study area. Since 1986, the percent occurrence and importance of sandlance in the diet of seals has decreased, reflecting an overall decrease in abundance of this prey species in waters adjacent to Cape Cod. In spite of fluctuations in abundance, and regional differences in the diet of seals throughout the study area, sandlance still comprised a minimum 55% of the total prey species of harbor seals throughout the study area.     

Novel and canine genotypes of Giardia duodenalis in harbor seals ( Phoca vitulina richardsi)

Feces of harbor seals (Phoca vitulina richardsi) and hybrid glaucous-winged/western gulls (Larus glaucescens / occidentalis) from Washington State's inland marine waters were examined for Giardia and Cryptosporidium spp. to determine if genotypes carried by these wildlife species were the same genotypes that commonly infect humans and domestic animals. Using immunomagnetic separation followed by direct fluorescent antibody detection, Giardia spp. cysts were detected in 42% of seal fecal samples (41/97). Giardia-positive samples came from 90% of the sites (9/10) and the prevalence of positive seal fecal samples differed significantly among study sites. Fecal samples collected from seal haulout sites with over 400 animals were 4.7 times more likely to have Giardia spp. cysts than samples collected at smaller haulout sites. In gulls, a single Giardia sp. cyst was detected in 4% of fecal samples (3/78). Cryptosporidium spp. oocysts were not detected in any of the seals or gulls tested. Sequence analysis of a 398 bp segment of G. duodenalis DNA at the glutamate dehydrogenase locus suggested that 11 isolates originating from seals throughout the region were a novel genotype and 3 isolates obtained from a single site in south Puget Sound were the G. duodenalis canine genotype D. Real-time TaqMan PCR amplification and subsequent sequencing of a 52 bp small subunit ribosomal DNA region from novel harbor seal genotype isolates showed sequence homology to canine genotypes C and D. Sequence analysis of the 52 bp small subunit ribosomal DNA products from the 3 canine genotype isolates from seals produced mixed sequences at could not be evaluated.     

Feeding habits of three seal species at the Danco Coast, Antarctica: a re-assessment

The analysis of prey overlap among Weddell, Antarctic fur and leopard seals was conducted using fecal samples collected at the Danco Coast, Antarctic Peninsula, in 1998 and 2000. The re-occurrence of prey species was moderate in samples collected in 1998, and low in 2000, and reflects resource partitioning among seal species. Prey species that mostly co-occurred in seals’ diet were the Antarctic krill Euphausia superba, bivalves, and the myctophids Gymnoscopelus nicholsi and Electrona antarctica. A dietary similarity index of prey overlap has been calculated and demonstrates evident fluctuations in pairwise comparisons between the seal species. The highest and lowest values of prey overlap were observed between Antarctic fur seals and leopard seals, and between Weddell seals and leopard seals, respectively. Prey overlap between Antarctic fur seals and Weddell seals was moderate in both seasons.     

Molecular scatology: the use of molecular genetic analysis to assign species, sex and individual identity to seal faeces

Seals and commercial fisheries are potential competitors for fish and cephalopods. Research into the diet of British seal species has been based on conventional dietary analyses, but these methods often do not allow assignment of species identity to scat samples. We present a protocol for obtaining DNA from seal scat (faecal) samples which can be used in polymerase chain reactions to amplify both nuclear and mitochondrial DNA. This can provide a method of identifying the species, sex and individual identity of the seal, from a particular scat sample. Combined with conventional dietary analyses these techniques will allow us to assess sources of variation in seal diet composition.
Scat samples have been collected from intertidal haul-out sites around the inner Moray Firth, north-east Scotland. We have assessed methods to extract and purify faecal DNA, a combination of DNA from the individual seal, prey items, and gut bacteria, for use in PCR. Controls using faecal and blood samples from the same individual have enabled microsatellite primer sets from four pinniped species to be tested. Approximately 200 scat samples have been examined for species identity and individual matches. This study will provide essential information for the assessment of interactions between seals and commercial or recreational fisheries.     

Environmental DNA from Residual Saliva for Efficient Noninvasive Genetic Monitoring of Brown Bears (Ursus arctos)

Noninvasive genetic sampling is an important tool in wildlife ecology and management, typically relying on hair snaring or scat sampling techniques, but hair snaring is labor and cost intensive, and scats yield relatively low quality DNA. New approaches utilizing environmental DNA (eDNA) may provide supplementary, cost-effective tools for noninvasive genetic sampling. We tested whether eDNA from residual saliva on partially-consumed Pacific salmon (Oncorhynchus spp.) carcasses might yield suitable DNA quality for noninvasive monitoring of brown bears (Ursus arctos). We compared the efficiency of monitoring brown bear populations using both fecal DNA and salivary eDNA collected from partially-consumed salmon carcasses in Southeast Alaska. We swabbed a range of tissue types from 156 partially-consumed salmon carcasses from a midseason run of lakeshore-spawning sockeye (O. nerka) and a late season run of stream-spawning chum (O. keta) salmon in 2014. We also swabbed a total of 272 scats from the same locations. Saliva swabs collected from the braincases of salmon had the best amplification rate, followed by swabs taken from individual bite holes. Saliva collected from salmon carcasses identified unique individuals more quickly and required much less labor to locate than scat samples. Salmon carcass swabbing is a promising method to aid in efficient and affordable monitoring of bear populations, and suggests that the swabbing of food remains or consumed baits from other animals may be an additional cost-effective and valuable tool in the study of the ecology and population biology of many elusive and/or wide-ranging species.     

Fish prey of Antarctic fur seals,Arctocephalus gazella,during the summer-autumn period at Laurie Island,South Orkney Islands

The fish component in the diet of Antarctic fur seals, Arctocephalus gazella, was investigated at Laurie Island, South Orkney Islands from mid January to April 1988. Fish otoliths occurred in 78.8% of faecal droppings. Most of the otoliths extracted from scats belonged to Myctophid fish (93.4%), mainly Electrona antarctica and Gymnoscopelus nicholsi. These two fish species thus constituted the bulk of the diet. A greater percentage of the otoliths from G. nicholsi (31.1%) in this study were considered suitable for measurement in comparison with 10.3% from scats at Heard Island in 1990 and 11.4% at Macquarie Island in 1988/89. This lesser degree of erosion would suggest that foraging areas of fur seals during this study were closer to land than during previous studies. During the period studied, the commercial fishery around the South Orkneys was not based on Myctophid fish so there was no direct competition for fish between the fur seals and fisheries.     

Evaluation of Scat Deposition Transects versus Radio Telemetry for Developing a Species Distribution Model for a Rare Desert Carnivore,the Kit Fox

Development and evaluation of noninvasive methods for monitoring species distribution and abundance is a growing area of ecological research. While noninvasive methods have the advantage of reduced risk of negative factors associated with capture, comparisons to methods using more traditional invasive sampling is lacking. Historically kit foxes (Vulpes macrotis) occupied the desert and semi-arid regions of southwestern North America. Once the most abundant carnivore in the Great Basin Desert of Utah, the species is now considered rare. In recent decades, attempts have been made to model the environmental variables influencing kit fox distribution. Using noninvasive scat deposition surveys for determination of kit fox presence, we modeled resource selection functions to predict kit fox distribution using three popular techniques (Maxent, fixed-effects, and mixed-effects generalized linear models) and compared these with similar models developed from invasive sampling (telemetry locations from radio-collared foxes). Resource selection functions were developed using a combination of landscape variables including elevation, slope, aspect, vegetation height, and soil type. All models were tested against subsequent scat collections as a method of model validation. We demonstrate the importance of comparing multiple model types for development of resource selection functions used to predict a species distribution, and evaluating the importance of environmental variables on species distribution. All models we examined showed a large effect of elevation on kit fox presence, followed by slope and vegetation height. However, the invasive sampling method (i.e., radio-telemetry) appeared to be better at determining resource selection, and therefore may be more robust in predicting kit fox distribution. In contrast, the distribution maps created from the noninvasive sampling (i.e., scat transects) were significantly different than the invasive method, thus scat transects may be appropriate when used in an occupancy framework to predict species distribution. We concluded that while scat deposition transects may be useful for monitoring kit fox abundance and possibly occupancy, they do not appear to be appropriate for determining resource selection. On our study area, scat transects were biased to roadways, while data collected using radio-telemetry was dictated by movements of the kit foxes themselves. We recommend that future studies applying noninvasive scat sampling should consider a more robust random sampling design across the landscape (e.g., random transects or more complete road coverage) that would then provide a more accurate and unbiased depiction of resource selection useful to predict kit fox distribution.     

Improved estimates of digestion correction factors and passage rates for harbor seal (Phoca vitulina) prey in the northeast Atlantic

Diet composition in pinnipeds is widely estimated using hard prey remains recovered from feces. To estimate the size and number of prey represented in fecal samples accurately, digestion correction factors (DCFs) must be applied to measurements and counts of fish otoliths and cephalopod beaks. In this study, 101 whole prey feeding trials were conducted with six harbor seals (Phoca vitulina) and 18 prey species. We derived species‐ and grade‐specific estimates of digestion coefficients (DCs) and species‐specific recovery rates (RRs) to account for partial and complete digestion, respectively. Greater than 98% of otoliths were passed within three days of consumption. RRs were smallest for Atlantic salmon smolts (RR = 0.306, SE = 0.031) and increasingly larger for sandeels (RR = 0.494, SE = 0.017), flatfish (RR = 0.789, SE = 0.033), and large gadoids (RR = 0.944, SE = 0.034). Species‐specific otolith width DCs were smallest for Trisopterus species (DC = 1.14, SE = 0.015) and increasingly larger for flatfish (DC = 1.27, SE = 0.045), large gadoids (DC = 1.32, SE = 0.067) and sandeels (DC = 1.57, SE = 0.035). RRs were similar to those from gray seals (Halichoerus grypus), but harbor seal species‐ and grade‐specific DCs were generally smaller. Differences in partial and complete digestion rates among prey species and between seal species highlight the importance of applying DCFs when reconstructing diet.     

Evaluation of fecal DNA preservation techniques and effects of sample age and diet on genotyping success

Optimal collection and preservation protocols for fecal DNA genotyping are not firmly established. We evaluated 3 factors that influence microsatellite genotyping success of fecal DNA extracted from coyote (Canis latrans) scats: 1) age of scat, 2) preservative, and 3) diet content. We quantified genotyping success by comparing rates of allelic dropout, false alleles, and failed amplifications among consensus genotypes. We used a panel of 6 microsatellite loci to genotype 20 scat samples, each of which was subjected to 3 age (1 day, 5 days, and 10 days post-deposition) and 3 preservation (DET buffer, 95% ethanol [EtOH], and lysis buffer) treatments. Both sample age and storage buffer had a significant effect on success and reliability. Ethanol and DET buffer preserved fecal samples with similar efficiency, and both were superior to lysis buffer. Our analysis of DNA degradation rates revealed that samples collected as early as 5 days of age yielded DNA that was highly degraded relative to samples collected on day 1. We tested the influence of dietary remains on microsatellite genotyping by using scat samples consisting predominantly of insect prey (n = 5), mammalian prey (n = 9), or the remains of juniper (Juniperus spp.) berries (n = 6) and compared EtOH and DET buffer preservation efficacy. We observed a significant interaction effect between storage buffer and diet for the probability of a false allele in a polymerase chain reaction (PCR), suggesting that the optimal preservation technique depended on the food remains comprising the scat. Scats comprised of juniper berry remains were more reliably genotyped when preserved in DET than EtOH. Mammalian prey-based scats were more reliable when stored in EtOH than DET buffer. Insect-predominant scats were preserved in EtOH and DET buffer with similar efficiency. Although accurate and reliable results can be obtained from scats collected at ≥5 days of age, we suggest sampling design to include collection of scats <5 days of age to minimize field and laboratory expenses. We suggest EtOH preservation for scats of obligate carnivores and of facultative carnivores with a diet consisting primarily of mammals. We suggest DET buffer preservation for animals with a diet consisting of plant-derived foods. Lysis buffer protocols that we employed should not be used for fecal DNA preservation. © 2011 The Wildlife Society.     

The diet of Weddell seals in McMurdo Sound, Antarctica as determined from scat collections and stable isotope analysis

The diet of adult and juvenile Weddell seals (Leptonychotes weddellii) in McMurdo Sound, Antarctica, was determined from both scat and stable isotope analyses, to ascertain if foraging behavior varied with age, season, or diving pattern. Scats were collected over 6 years and recovered hard parts identified. Stable carbon and nitrogen isotope values were determined for seal blood samples and potential prey items and used to identify primary prey species and assess trophic interactions. Pleuragramma antarcticum remains were recovered from between 70 and 100% of the scats, and there was little evidence for inter-annual or age-specific variation in foraging behavior. However, stable isotope and dive data analyses indicated that while most seals foraged predominantly on pelagic fish and squid, some juveniles concentrated on shallow benthic Trematomus spp. Combining these three methods permitted firm conclusions about diet and foraging behavior to be drawn. Received: 10 June 1997 / Accepted: 8 November 1997