Detection distance and environmental factors in conservation detection dog surveys

Surveys using conservation detection dogs have grown increasingly popular as an efficient means to gather monitoring data, particularly for elusive and low-density species such as carnivores. Working with dogs can greatly increase the area surveyed for wildlife and the detection rate of survey targets. Due to the confounding effects of scent dispersion and dog movement, however, it can be difficult to estimate the area searched in a survey. Additionally, although detection dogs have been used in studies under a wide range of air temperature, humidity, and wind conditions, little research has examined how environmental factors affect detection dogs' effectiveness for wildlife surveys. Between 2003 and 2005, we trained 2 dogs to assist us with surveys for mammalian carnivore scats in northern California. We conducted controlled search trials to assess how the dogs' scat detection rates were affected by the distance of scats from the transect search line, as well as variation in six environmental factors. Both dogs detected >75% of scats located within 10 m, and the dogs' detection rates decreased with increasing distance of scats from the transect line. Among environmental factors, precipitation was the most important variable explaining variation in scat detection rates for both dogs. Precipitation likely degrades or removes scats from the landscape over time, and detection rates increase as scat begins to accumulate following the last substantial (>5 mm) rain event of the year. If scat accumulation is not controlled for in ecosystems with a strong seasonal pattern of rainfall, it could lead to considerable bias in study results. We recommend that researchers report the conditions under which conservation detection dog surveys took place and analyze how detection rates vary as a function of distance, temperature, precipitation, humidity, wind, and other locally important environmental factors. © 2010 The Wildlife Society.     

Use of Volatile Organic Components in Scat to Identify Canid Species

Abstract: Identification of wildlife species from indirect evidence can be an important part of wildlife management, and conventional methods can be expensive or have high error rates. We used chemical characterization of the volatile organic constituents (VOCs) in scat as a method to identify 5 species of North American canids from multiple individuals. We sampled vapors of scats in the headspace over a sample using solid-phase microextraction and determined VOC content using gas chromatography with a flame ionization detector. We used linear discriminant analysis to develop models for differentiating species with bootstrapping to estimate accuracy. Our method correctly classified 82.4% (bootstrapped 95% CI = 68.8–93.8%) of scat samples. Red fox (Vulpes vulpes) scat was most frequently misclassified (25.0% of scats misclassified); red fox was also the most common destination for misclassified samples. Our findings are the first reported identification of animal species using VOCs in vapor emissions from scat and suggest that identification of wildlife species may be plausible through chemical characterization of vapor emissions of scat. (JOURNAL OF WILDLIFE MANAGEMENT 72(3):792–797; 2008)     

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.     

An Improved Field Method to Obtain DNA for Individual Identification From Wolf Scat

ABSTRACT Sampling of feces for genetic studies of wild populations can be problematic because of the low quality and quantity of template DNA obtained. We used cotton swabs in the field to isolate the mucous layer on the surface of fresh wolf (Canis lupus, C. lycaon, and their hybrids) scats followed by immediate preservation, and compared microsatellite genotyping of DNA from these fresh field swabs (FS) to that of previously frozen laboratory swabs (LS). In single polymerase chain reactions (PCRs) of 2 multiplexes, amplification at 8 loci was higher in the FS samples (FS = 50%, LS = 15%; P = 0.02) because proportion, quantity, and quality of large fragment wolf nuclear DNA from these samples was greater (2.5–25%, 6.25–62.5 ng/swab, 35% amplified at 1,000 base pairs [bp]) than from the LS samples (1.9%–10%, 4.7–25 ng/swab, 10% amplified at 1,000 bp). Paired blood and fresh field-swabbed samples had identical genotypes. In 84 multiplex PCRs we found no evidence of allelic dropout associated with low template quality or quantity. We conclude that field swabbing of fresh wolf scat facilitates field storage and reduces the need for multiple amplifications at single microsatellite loci, thereby reducing the genotyping costs for wildlife projects that use noninvasive samples.     

Non-invasive genetic identification of small mammal species using real-time polymerase chain reaction

DNA identification of non-invasive samples is a potentially useful tool for monitoring small mammal species. Here we describe a novel method for identifying five small mammal species: wood mouse, bank vole, common shrew, pygmy shrew and water shrew. Species-specific real-time polymerase chain reaction primers were designed to amplify fragments of the mitochondrial cytochrome b gene from hair and scat samples. We also amplified nuclear DNA from scats, demonstrating their potential as a source of DNA for population genetic studies.     

Developing noninvasive methodologies to assess koala population health through detecting Chlamydia from scats

Wildlife diseases are a recognized driver of global biodiversity loss, have substantial economic impacts, and are increasingly becoming a threat to human health. Disease surveillance is critical but remains difficult in the wild due to the substantial costs and potential biases associated with most disease detection methods. Noninvasive scat surveys have been proposed as a health monitoring methodology to overcome some of these limitations. Here, we use the known threat of Chlamydia disease to the iconic, yet vulnerable, koala Phascolarctos cinereus to compare three methods for Chlamydia detection in scats: multiplex quantitative PCR, next generation sequencing, and a detection dog specifically trained on scats from Chlamydia‐infected koalas. All three methods demonstrated 100% specificity, while sensitivity was variable. Of particular interest is the variable sensitivity of these diagnostic tests to detect sick individuals (i.e., not only infection as confirmed by Chlamydia‐positive swabs, but with observable clinical signs of the disease); for koalas with urogenital tract disease signs, sensitivity was 78% with quantitative PCR, 50% with next generation genotyping and 100% with the detection dog method. This may be due to molecular methods having to rely on high‐quality DNA whereas the dog most likely detects volatile organic compounds. The most appropriate diagnostic test will vary with disease prevalence and the specific aims of disease surveillance. Acknowledging that detection dogs might not be easily accessible to all, the future development of affordable and portable “artificial noses” to detect diseases from scats in the field might enable cost‐effective, rapid and large‐scale disease surveillance.     

Changes in Kit Fox Defecation Patterns During the Reproductive Season: Implications for Noninvasive Surveys

Abstract: Noninvasive survey methods based on analyzing DNA extracted from feces can be useful for carnivores that are difficult to study by other methods. Changes in fecal deposition patterns associated with reproduction in kit foxes (Vulpes macrotis) might affect results of such surveys. We used a trained dog to collect fresh scats on 2-km transects in the home ranges of 11 radiocollared female kit foxes in January, February, and March 2008 and determined sex of the individual that deposited the scats by amplifying the zinc finger protein gene. Female foxes give birth in mid-February to mid-March. We found a similar number of scats each month. In January, the sex ratio of the scats was not different from the expected 1:1. However, in February there were almost 2 male scats for every female scat and in March there were >8 male scats for every female scat. Comparing March to January, there were more male scats on all 11 transects and fewer female scats on 10 of 11 transects. Around the time pups are born, both sexes appear to show changes in fecal deposition patterns that make it easier to find male scats and harder to find female scats. Effects of these changes on survey results will vary depending on the purpose and design of the survey. Surveys to determine distribution and relative abundance would probably not be negatively affected by these changes. However, if surveys to estimate abundance are conducted during the reproductive season, they could result in an underestimate of population size unless the increased heterogeneity in scat detectability is taken into account.     

Multiple species‐specific molecular markers using nanofluidic array as a tool to detect prey DNA from carnivore scats

Large carnivore feeding ecology plays a crucial role for management and conservation for predators and their prey. One of the keys to this kind of research is to identify the species composition in the predator diet, for example, prey determination from scat content. DNA‐based methods applied to detect prey in predators’ scats are viable alternatives to traditional macroscopic approaches, showing an increased reliability and higher prey detection rate. Here, we developed a molecular method for prey species identification in wolf (Canis lupus) scats using multiple species‐specific marker loci on the cytochrome b gene for 18 target species. The final panel consisted of 80 assays, with a minimum of four markers per target species, and that amplified specifically when using a high‐throughput Nanofluidic array technology (Fluidigm Inc.). As a practical example, we applied the method to identify target prey species DNA in 80 wolf scats collected in Sweden. Depending on the number of amplifying markers required to obtain a positive species call in a scat, the success in determining at least one prey species from the scats ranged from 44% to 92%. Although we highlight the need to evaluate the optimal number of markers for sensitive target species detection, the developed method is a fast and cost‐efficient tool for prey identification in wolf scats and it also has the potential to be further developed and applied to other areas and large carnivores as well.     

Dietary separation of sympatric carnivores identified by molecular analysis of scats

We studied the diets of four sympatric carnivores in the flooding savannas of western Venezuela by analysing predator DNA and prey remains in faeces. DNA was isolated and a portion of the cytochrome b gene of the mitochondrial genome amplified and sequenced from 20 of 34 scats. Species were diagnosed by comparing the resulting sequences to reference sequences generated from the blood of puma (Puma concolor), jaguar (Panthera onca), ocelot (Leopardus pardalus) and crab-eating fox (Cerdocyon thous). Scat size has previously been used to identify predators, but DNA data show that puma and jaguar scats overlap in size, as do those of puma, ocelot and fox. Prey-content analysis suggests minimal prey partitioning between pumas and jaguars. In field testing this technique for large carnivores, two potential limitations emerged: locating intact faecal samples and recovering DNA sequences from samples obtained in the wet season. Nonetheless, this study illustrates the tremendous potential of DNA faecal studies. The presence of domestic dog (Canis familiaris) in one puma scat and of wild pig (Sus scrofa), set as bait, in one jaguar sample exemplifies the forensic possibilities of this noninvasive analysis. In addition to defining the dietary habits of similar size sympatric mammals, DNA identifications from faeces allow wildlife managers to detect the presence of endangered taxa and manage prey for their conservation.     

Molecular scatology as a tool to study diet: analysis of prey DNA in scats from captive Steller sea lions

The DNA of prey present in animal scats may provide a valuable source of information for dietary studies. We conducted a captive feeding trial to test whether prey DNA could be reliably detected in scat samples from Steller sea lions (Eumetopias jubatus). Two sea lions were fed a diet of fish (five species) and squid (one species), and DNA was extracted from the soft component of collected scats. Most of the DNA obtained came from the predator, but prey DNA could be amplified using prey-specific primers. The four prey species fed in consistent daily proportions throughout the trial were detected in more than 90% of the scat DNA extractions. Squid and sockeye salmon, which were fed as a relatively small percentage of the daily diet, were detected as reliably as the more abundant diet items. Prey detection was erratic in scats collected when the daily diet was fed in two meals that differed in prey composition, suggesting that prey DNA is passed in meal specific pulses. Prey items that were removed from the diet following one day of feeding were only detected in scats collected within 48 h of ingestion. Proportions of fish DNA present in eight scat samples (evaluated through the screening of clone libraries) were roughly proportional to the mass of prey items consumed, raising the possibility that DNA quantification methods could provide semi-quantitative diet composition data. This study should be of broad interest to researchers studying diet since it highlights an approach that can accurately identify prey species and is not dependent on prey hard parts surviving digestion.     

Balancing sample accumulation and DNA degradation rates to optimize noninvasive genetic sampling of sympatric carnivores

Noninvasive genetic sampling, or noninvasive DNA sampling (NDS), can be an effective monitoring approach for elusive, wide‐ranging species at low densities. However, few studies have attempted to maximize sampling efficiency. We present a model for combining sample accumulation and DNA degradation to identify the most efficient (i.e. minimal cost per successful sample) NDS temporal design for capture–recapture analyses. We use scat accumulation and faecal DNA degradation rates for two sympatric carnivores, kit fox (Vulpes macrotis) and coyote (Canis latrans) across two seasons (summer and winter) in Utah, USA, to demonstrate implementation of this approach. We estimated scat accumulation rates by clearing and surveying transects for scats. We evaluated mitochondrial (mtDNA) and nuclear (nDNA) DNA amplification success for faecal DNA samples under natural field conditions for 20 fresh scats/species/season from <1–112 days. Mean accumulation rates were nearly three times greater for coyotes (0.076 scats/km/day) than foxes (0.029 scats/km/day) across seasons. Across species and seasons, mtDNA amplification success was ≥95% through day 21. Fox nDNA amplification success was ≥70% through day 21 across seasons. Coyote nDNA success was ≥70% through day 21 in winter, but declined to <50% by day 7 in summer. We identified a common temporal sampling frame of approximately 14 days that allowed species to be monitored simultaneously, further reducing time, survey effort and costs. Our results suggest that when conducting repeated surveys for capture–recapture analyses, overall cost‐efficiency for NDS may be improved with a temporal design that balances field and laboratory costs along with deposition and degradation rates.     

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.     

基于粪便DNA 的青海雪豹种群遗传结构初步研究

雪豹是国际关注的全球濒危物种,由于独特的生活习性,其确切分布区、种群数量和遗传结构信息非常有限,基于粪便DNA 分析技术的发展为该物种的研究提供了新的手段。在青海省都兰县宗加乡、诺木红乡和治多县索加乡收集到106 份疑似雪豹粪便样品,成功地扩增了78 份粪便样品的mtDNA Cyt b 基因片段,并进行测序。经过GenBank 数据库的Blastn 比对,确定了21 份为雪豹粪便,其中宗加乡11 份、诺木红乡5 份、索加乡5份。经过Clustal W 和DNASP 软件比对分析,在21 份雪豹粪便DNA 中,共检测出7 个多态性位点,分为4 个单倍型,单倍型多态性(h)为0. 384,核苷酸多态性(π)为0.35% 。三个样地之间的遗传距离为0.002 ～ 0.005,核苷酸差异为0.200 ～1. 273。结果表明了3 个取样区均有雪豹存在,且存在较丰富的遗传多态性,样地间也存在一定的遗传差异。     

Ageing and environmental factors affect PCR success in wolf (Canis lupus) excremental DNA samples

Individual genotypes determined from noninvasive DNA samples (typically extracted from shed hairs or scats) are used to estimate population size in monitoring projects of elusive species. However, polymerase chain reaction (PCR) success rates usually are lower, and genotyping errors higher than in standard population genetic surveys, due to DNA degradation or contamination in aged field samples. In this study, we evaluate the results of common garden experiments showing that DNA degradation is significant in wolf (Canis lupus) scats older than 3 days, and it is enhanced in scats in direct contact with soil. A storage test showed that samples kept frozen in 95% ethanol performed better compared to other methods. However, variance of PCR success among samples was high, independent on sample age or storage condition. The detrimental consequences of DNA degradation can be avoided by collecting scat samples as fresh as possible, and implementing efficient multitube procedures and stringent quality control of the laboratory results. Efficient multitube procedures can produce reliable data, like in this study, which showed that the consensus genotypes obtained from excremental DNA exactly matched distinct reference genotypes obtained from wolf blood samples.     

Amplification of DNA markers from scat samples of the least weasel<Emphasis Type="Italic">Mustela nivalis nivalis</Emphasis>

To test the feasibility of using field-collected scats as a source of DNA in the study of the least weaselMustela nivalis nivalis Linnaeus, 1766, DNA was extracted from scat samples collected from captive weasels using a modified extraction protocol. Using universal primers, the control region of the mitochondrial genome was successfully amplified from scat-extracted DNA. This amplification resulted in two products; one equivalent in size and sequence to the product obtained from tissue-extracted weasel DNA, and the other slightly larger and equivalent in size and sequence to the domestic house mouseMus musculus, the food source of the captive weasels. This demonstrates the reliability of DNA extraction from scats, as well as the possibility, under favourable circumstances, of identifying the prey species from the same samples. In addition, we attempted to amplify microsatellite loci from both tissue and scat-extracted DNA using six primer pairs designed for other mustelids, the American minkMustela vison and the wolverineGulo gulo. While three loci, Mvi57 (American mink), Ggu216 and Ggu234 (wolverine), were found to be polymorphic in the least weasel, amplification of these loci from the scat extracted DNA was only successful for approximately half of the samples. Although further work is needed, the present results suggest that it is possible to use scats as a source of DNA in field studies of the least weasel.     

Dispersal of Spores of Mycorrhizal Fungi in Scats of Native Mammals in Tropical Forests of Northeastern Australia

One hundred and thirty-eight scat (faecal) samples from 17 mammal species native to forests of northeastern Queensland were examined for the presence of spores of both ectomycorrhizal and arbuscular mycorrhizal fungi. Spores of mycorrhizal fungi were found in 57 percent of scat samples representing 12 animal species (Aepyprymnus rufescens, Antechinus godmani, Bettongia tropica, Hypsiprymnodon moschatus, Isoodon macrourus, Melomys ceruinipes, Perameles nasuta, Rattus fuscipes, R. tunneyi, Thylogale stigmatica, Trichourur uulperula, Uromys caudimaculatus). Spores were absent in scats of Antechinus stuartii, Dasyurus hallucatus, Dendrolagus lumholtzi, Petaurus australis and Mesembriomys gouldii. Spores of ectomycorrhizal fungi occurred in 38 percent of scats, and all but one of these samples were from Eucalyptus-dominated sclerophyll forests. Based on the frequency and abundance of spores in scats, five mammals were considered active consumers of hypogeous mycorrhizal sporocarps in sclerophyll forests (A. rufescens, B. tropica, I. macrourus, P. nasuta, and U. caudimaculatus). Individual scats of these animals generally contained a range of distinctive spore types. Spores of arbuscular mycorrhizal fungi were found in low abundance in almost 40 percent of scat samples collected, from both sclerophyll forest and rainforest habitats. We suggest that the majoriry of these spores were acquired incidentally through ingestion of soil during foraging activities on the forest floor. Glasshouse inoculation experiments in which seedlings of Eucalyptus grandis and Sorghum bicolor were inoculated with scat material from several species of mammal demonstrated that the spores of ectomycorrhizal and arbuscular mycorrhizal fungi retained some viability and colonized the roots of host-plant seedlings. Insufficient information is known of the ecology of mycorrhizal fungi in Australia's tropical forests to speculate as to the implications of these findings for forest conservation and rehabilitation.     

Assessing the effectiveness of long-term monitoring of the Broad-toothed Rat in the Barrington Tops National Park,Australia

Biodiversity monitoring is crucial for effective conservation efforts. Effective monitoring allows managers to determine the status and trends of biodiversity, as well as the success of conservation actions. The population of the Broad-toothed Rats (Mastacomys fuscus) in the Barrington Tops National Park New South Wales, Australia has been monitored since 1999 via scat and live-trapping surveys. We reviewed the methods used and analysed the data produced with the aim of describing patterns of population change over time using a range of outcome variables and identifying different climate correlates. A secondary aim was to explore the use of population statistics that account for imperfect detection by comparing naïve occupancy, with an index of relative abundance based on trap effort, the latency to find scats during scat surveys and an occupancy model based on trapping surveys. Neither of these three methods accounts for detectability variation. Naïve occupancy decreased slightly over time, while the relative abundance based on trap effort revealed no evidence of change. Additionally, naïve occupancy decreased with increasing temperature while temperature had no clear impact on relative abundance. Finally, precipitation had no impact on either naïve occupancy or relative abundance. We found no evidence of a relationship between the latency to find scats and the index of relative abundance, suggesting that one or neither is related to actual abundance. Finally, a multi-season occupancy model found occupancy probability to be 0.78 ± 0.23 (standard error); detection probability as 0.51 ± 0.06; seasonal colonisation rate as 0.36 ± 0.13 and seasonal extinction rate at 0.44 ± 0.13. We conclude that despite significant investment in monitoring, this historical data set does not allow managers to ascertain whether population change has occurred and to identify potential drivers of change. Careful consideration of future methods, in particular, whether there is imperfect detection in scat surveys, will help to inform future monitoring.     

Scent-Matching Dogs Determine Number of Unique Individuals From Scat

ABSTRACT Noninvasive scat sampling methods can generate large samples sizes, collected over vast landscapes, ideal for addressing wildlife conservation and management questions. However, the cost of genotyping scat samples limits the accessibility of these techniques. We describe detection-dog methods for matching large numbers of scat samples to the individual, reducing or eliminating the need for sample genotyping. Three dogs correctly matched 25 out of 28 samples from 6 captive maned wolves (Chrysocyon brachyurus) of known identity. Sample scent-matching can increase overall accessibility and breadth of applications of noninvasive scat-collection methods to important landscape scale problems in wildlife sciences.     

River otter population size estimation using noninvasive latrine surveys

Across much of North America, river otter (Lontra canadensis) populations were extirpated or greatly reduced by the early 20th century. More recently, reintroductions have resulted in restored populations and the recommencement of managed trapping. Perhaps the best example of these river otter reintroductions occurred in Missouri, regarded as one of the most successful carnivore recovery programs in history. However, abundance estimates for river otter populations are difficult to obtain and often contentious when used to underpin management activities. We assessed the value of latrine site monitoring as a mechanism for quantifying river otter abundance. Analyses of fecal DNA to identify individual animals may result in an improved population estimate and have been used for a variety of mammal species. We optimized laboratory protocols, redesigned existing microsatellite primers, and calculated genotyping error rates to enhance genotyping success for a large quantity of river otter scat samples. We also developed a method for molecular sexing. We then extracted DNA from 1,421 scat samples and anal sac secretions (anal jelly) collected during latrine site counts along 22–34-km stretches representing 8–77% of 8 rivers in southern Missouri in 2009. Error rates were low for the redesigned microsatellites. We obtained genotypes at 7–10 microsatellite loci for 24% of samples, observing highest success for anal jelly samples (71%) and lowest for fresh samples (collected within 1 day of defecation). We identified 63 otters (41 M, 22 F) in the 8 rivers, ranging from 2 to 14 otters per river. Analyses using program CAPWIRE resulted in population estimates similar to the minimum genotyping estimate. Density estimates averaged 0.24 otters/km. We used linear regression to develop and contrast models predicting population size based on latrine site and scat count indices, which are easily collected in the field. Population size was best predicted by a combination of scats per latrine and latrines per kilometer. Our results provide methodological approaches to guide wildlife managers seeking to initiate similar river otter fecal genotyping studies, as well as to estimate and monitor river otter population sizes. © 2011 The Wildlife Society.     

Efficient species identification of pine marten (<Emphasis Type="Italic">Martes martes</Emphasis>) and red fox (<Emphasis Type="Italic">Vulpes vulpes</Emphasis>) scats using a 5′ nuclease real-time PCR assay

Monitoring wildlife species by DNA identification of samples collected non-invasively is an important tool in conservation management. DNA identification of species from faecal (scat) samples is problematic due to the small quantities and poor quality of the DNA isolated from such samples. This study demonstrates the use of real-time PCR technology in the identification of red fox (Vulpes vulpes) and pine marten (Martes martes). It is shown that real-time PCR can be used to identify fox and pine marten by either melting curve analysis (Tm determination) with SYBR Green 1 detection or by the use of species specific fluorogenic probes. The technique is shown to work efficiently with scat DNA.