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.     

Impacts of sampling location within a faeces on DNA quality in two carnivore species

We investigated the influence of sampling location within a faeces on DNA quality by sampling from both the outside and inside of 25 brown bear (Ursus arctos) scats and the side and the tip of 30 grey wolf (Canis lupus) scats. The outside of the bear scat and side of the wolf scat had significantly lower nuclear DNA microsatellite allelic dropout error rates (U. arctos: P = 0.017; C. lupus: P = 0.025) and significantly higher finalized genotyping success rates (U. arctos: P = 0.017; C. lupus: P = 0.012) than the tip and inside of the scat. A review of the faecal DNA literature indicated that <45% of studies report the sampling location within a faeces indicating that this methodological consideration is currently underappreciated. Based on our results, we recommend sampling from the side of canid scats and the outside portion of ursid scats to obtain higher quality DNA samples. The sampling location within a faeces should be carefully considered and reported as it can directly influence laboratory costs and efficiency, as well as the ability to obtain reliable genotypes.     

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.     

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.     

An efficient method for screening faecal DNA genotypes and detecting new individuals and hybrids in the red wolf (<Emphasis Type="Italic">Canis rufus</Emphasis>) experimental population area

Previously, sequencing of mitochondrial DNA (mtDNA) from non-invasively collected faecal material (scat) has been used to help manage hybridization in the wild red wolf (Canis rufus) population. This method is limited by the maternal inheritance of mtDNA and the inability to obtain individual identification. Here, we optimize the use of nuclear DNA microsatellite markers on red wolf scat DNA to distinguish between individuals and detect hybrids. We develop a data filtering method in which scat genotypes are compared to known blood genotypes to reduce the number of PCR amplifications needed. We apply our data filtering method and the more conservative maximum likelihood ratio method (MLR) of Miller et al. (2002 Genetics 160:357–366) to a scat dataset previously screened for hybrids by sequencing of mtDNA. Using seven microsatellite loci, we obtained genotypes for 105 scats, which were matched to 17 individuals. The PCR amplification success rate was 50% and genotyping error rates ranged from 6.6% to 52.1% per locus. Our data filtering method produced comparable results to the MLR method, and decreased the time and cost of analysis by 25%. Analysis of this dataset using our data filtering method verified that no hybrid individuals were present in the Alligator River National Wildlife Refuge, North Carolina in 2000. Our results demonstrate that nuclear DNA microsatellite analysis of red wolf scats provides an efficient and accurate approach to screen for new individuals and hybrids.     

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.     

Fresh is best: Accurate SNP genotyping from koala scats

Maintaining genetic diversity is a crucial component in conserving threatened species. For the iconic Australian koala, there is little genetic information on wild populations that is not either skewed by biased sampling methods (e.g., sampling effort skewed toward urban areas) or of limited usefulness due to low numbers of microsatellites used. The ability to genotype DNA extracted from koala scats using next‐generation sequencing technology will not only help resolve location sample bias but also improve the accuracy and scope of genetic analyses (e.g., neutral vs. adaptive genetic diversity, inbreeding, and effective population size). Here, we present the successful SNP genotyping (1272 SNP loci) of koala DNA extracted from scat, using a proprietary DArTseq^? protocol. We compare genotype results from two‐day‐old scat DNA and 14‐day‐old scat DNA to a blood DNA template, to test accuracy of scat genotyping. We find that DNA from fresher scat results in fewer loci with missing information than DNA from older scat; however, 14‐day‐old scat can still provide useful genetic information, depending on the research question. We also find that a subset of 209 conserved loci can accurately identify individual koalas, even from older scat samples. In addition, we find that DNA sequences identified from scat samples through the DArTseq^? process can provide genetic identification of koala diet species, bacterial and viral pathogens, and parasitic organisms.     

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.     

Procedures to genotype problematic non-invasive otter (Lutra lutra) samples

Non-invasive genetics is a powerful tool in wildlife research and monitoring, especially when dealing with elusive and rare species such as the Eurasian otter (Lutra lutra). Nevertheless, otter DNA obtained from scats and anal secretions appears to be exposed to very quick degradation processes, and the success rate in DNA amplification is lower than in other carnivores. We collected 191 samples from April to September 2011 along the river Sangro basin (Italy) which was recently re-colonized by the Eurasian otter. Using two sets of microsatellite loci (six Lut and seven OT loci), we investigated the influence of sample type and age, collection time, storage time, temperature and humidity on genotyping success and amplification success. We also tested the efficacy of different DNA extraction kits and storage buffer mediums. Finally, we compared amplification success rate, allelic dropout and false allele rates for each locus. We obtained a mean amplification success rate of 79.0 % and a genotyping success rate of 35.1 %. Fresh pure jellies yielded the highest amplification success and genotyping rate. Six microsatellite loci should be theoretically sufficient to distinguish the individual unrelated otters (PID?=?0.001), while 13 loci were needed to distinguish sibling otters (PID_sibs?=?0.002) in our population. We identified 11 otters, and molecular sexing ascertained the presence of five males, four females and two uncertain individuals. Generalized linear models highlighted a significant influence of sample type and age, temperature and humidity both on genotyping and amplification success.     

A comparison and critique of different scat‐analysis methods for determining carnivore diet

• 1 For terrestrial carnivores, scat analysis is the technique most often used to determine diets. Various methods of interpreting scat‐analysis data exist; however, little is known about how the choice of method affects the results.
• 2 We reviewed 50 scat‐analysis papers to assess the range of methods currently used. Furthermore, we used a large data set from cape fox Vulpes chama and black‐backed jackal Canis mesomelas scats to compare 11 scat‐analysis methods. Techniques tested included five biomass calculation methods, four frequency of occurrence methods, one method that estimated volume in scats, and another that estimated mass of food items in scats.
• 3 Frequency of occurrence methods were used in 94% of reviewed papers, and in 50% of papers they were the sole methods used. However, we conclude that frequency of occurrence has the least ecological significance and results can be misleading. Although biomass calculations probably provide the best approximation to true diets, only 23% of reviewed papers used suitable biomass calculation methods when models were available for the study species.
• 4 Analysis of fox and jackal scats showed that there were significant differences among methods when calculating percent diet composition and niche breadth. Additionally, dietary overlap between species differed considerably among the methods (range of R₀ = 0.29–0.79). We conclude that the choice of method can have a significant impact on the results of dietary analysis, and can lead to very different conclusions about a species' ecology.
• 5 The best approximation of the true diet can be obtained by using a biomass calculation model that was developed for the same species, or for a closely related species with a similar food spectrum. When no such model is available, either the volume or mass of diet components in the scats should be used. To document rare food items, frequency of occurrence data could also be given.

     

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.     

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.     

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.     

Evaluation of Noninvasive Genetic Sampling Methods for Felid and Canid Populations

Abstract: Noninvasive sampling methods provide a means for studying species such as large mammalian carnivores that are difficult to survey using traditional techniques. Focusing on bobcat (Lynx rufus), we compared the effectiveness of noninvasive hair and scat genetic sampling in terms of field sample collection, species identification, and individual identification. We describe a novel hair-snare design and sampling protocol that successfully sampled 4 sympatric carnivore species, bobcat, mountain lion (Felis concolor), coyote (Canis latrans), and gray fox (Urocyon cinereoargenteus), in 3 habitat blocks in coastal southern California, USA. Scat surveys were also successful at sampling bobcats and other carnivores in the area. Hair and scat sampling methods had similar species identification success (81% and 87%, respectively) using mitochondrial DNA amplification and restriction enzyme digestion patterns. Therefore, for studies focused on the distribution and activity of a suite of carnivore species, we recommend a combination of noninvasive methodologies, for example, targeting hair and scat surveys toward species and sites where they are most effective. Because of a higher success rate for scat (85%) than for hair samples (10%) when using 4 microsatellite loci and a multiple-tubes approach to verify individual genotypes, we suggest scat sampling is a better choice for studies that require individual identification of bobcats.     

Preservation of corals in salt-saturated DMSO buffer is superior to ethanol for PCR experiments

Specimen collection is time consuming and expensive, yet few laboratories test preservation methods before setting out on field expeditions. The most common preservation buffer used for coral specimens is >70% EtOH. However, alternatives exist that are less flammable, easier to ship, and are widely used in other taxa. Here, we compare the effects of salt-saturated DMSO (SSD) and EtOH preservation buffers on post-extraction DNA quantity and quality. We found that soft tissue integrity was better maintained and higher quantities of DNA were extracted from EtOH-preserved specimens; however, by all other measures, SSD was a superior preservative to EtOH. Extractions of SSD-preserved specimens resulted in higher molecular weight DNA, higher PCR success, and more efficient amplification than specimens preserved in EtOH. Our results show that SSD is generally a superior preservative to EtOH for specimens destined for PCR studies, but species-specific differences indicate that preservation comparisons should be undertaken before collection and storage of samples.     

Low Genotyping Error Rates and Noninvasive Sampling in Bighorn Sheep

Abstract Noninvasive DNA sampling allows studies of natural populations without disturbing the target animals. Unfortunately, high genotyping error rates often make noninvasive studies difficult. We report low error rates (0.0–7.5%/locus) when genotyping 18 microsatellite loci in only 4 multiplex polymerase chain reaction amplifications using fecal DNA from bighorn sheep (Ovis canadensis). The average locus-specific error rates varied significantly between the 2 populations (0.13% vs. 1.6%; P < 0.001), as did multi-locus genotype error rates (2.3% vs. 14.1%; P < 0.007). This illustrates the importance of quantifying error rates in each study population (and for each season and sample preservation method) before initiating a noninvasive study. Our error rates are among the lowest reported for fecal samples collected noninvasively in the field. This and other recent studies suggest that noninvasive fecal samples can be used in species with pellet-form feces for nearly any study (e.g., of population structure, gene flow, dispersal, parentage, and even genome-wide studies to detect local adaptation) that previously required high-quality blood or tissue samples.     

Towards more efficient large-scale DNA-based detection of terrestrial mammal predators from scats

The DNA detection of wildlife from environmental samples has the potential to contribute significantly to wildlife management and ecological research. In terrestrial ecosystems, much work has focused on the identification of mammal predators from faecal (scat) samples. However, the relatively high time and financial costs of collecting and analysing scat DNA remain barriers to more widespread implementation of such DNA detection methods, especially for high-throughput surveys. Here, we evaluate methods used for DNA extraction from scats, as applied to detection of the Australian red fox, an introduced predator. We compare the relative costs of two approaches: the method previously used to screen thousands of scat samples in surveys over several years, and a modified version which involves swabbing scats at the time of collection and using a mechanised liquid handling platform to extract DNA from the swabs. We demonstrate that mechanised DNA extraction from swabs is more efficient than manual DNA extraction from whole scats, in terms of both time and resources. This provides a means for rapid, high-throughput screening of scats for the presence of mammal predators, enabling time-effective management responses to non-invasive surveys.     

Prey preference of large carnivores in Anamalai Tiger Reserve,India

Prey preferences of large carnivores (tiger (Panthera tigris), leopard (Panthera pardus) and dhole (Cuon alpinus)) in the tropical forest of Anamalai Tiger Reserve (ATR) were evaluated. This was the first study in ATR to estimate the density of prey and the food habits of these large carnivores. The 958-km² intensive study area was found to have a high mammalian prey density (72.1 animals per square kilometre) with wild boar (20.61 animals per square kilometre) and chital (20.54 animals per square kilometre) being the most common species, followed by nilgiri tahr (13.6 animals per square kilometre). When the density figures were multiplied by the average weight of each prey species, a high biomass density of 14,204 kg km⁻² was obtained for the intensive study area. Scat analysis and incidental kill observation were used to determine the dietary composition of these predators. During the study from the period of March 2001 to April 2004, 1,145 tiger scats, 595 leopard scats and 2,074 dhole scats were collected and analysed. Kill data were based on direct observation of 66 tiger kills and 39 leopard kills. Sambar, with a density of 6.54 kg km⁻² was the preferred prey for these carnivores. Sambar constitutes 35% of the overall diet of tiger, whereas it constitutes 17% and 25% in leopard and dhole diets, respectively. Chital was utilized less than sambar in the range of about 7%, 11% and 15% by tiger, leopard and dhole, respectively. Predator diet was estimated more accurately by scat analysis, which reveals 30% of smaller prey species in leopard’s diet, which was not observed by kill data. This study reveals that ATR harbours high prey density, and these large carnivores seem mostly dependent on the wild prey rather than on domestic livestock as in some other areas in the subcontinent. These factors make ATR a potential area for long-term conservation of these endangered carnivores.     

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.     

Non-dietary analytical features of chimpanzee scats

Non-dietary aspects of ape scats such as scat weight and diameter are correlated with age and sex of defaecator for gorillas and orangutans. Defaecation rates of primates, including apes, illuminate their role as primary seed dispersers. We assess if non-dietary features of scats for East African chimpanzees (Pan troglodytes schweinfurthii) reveal such insights for members of the Kanyawara community in Kibale National Park, Uganda. Our objective is to see if such data yield useful perspectives for future census work on unhabituated chimpanzees, that is, what can scats tell us about a wild study population, beyond diet? We followed ten adults from this community, as well as travelling parties, comparing observed vs. unobserved defaecations, and collected data on scat weight and dimensions, defaecation rate, scat encounter rate, and interval between defaecations. Few non-dietary features of chimpanzee scats significantly differentiated sex or age of the defaecator, but total scat length and height distinguished adults from juveniles/infants. Defaecation rates and distance travelled were similar for adult males and females, indicating the importance of both sexes as potential primary seed dispersers. Observed travelling parties vs. non-observed travelling parties yielded similar data, indicating the potential to assess party size from scat encounter rates over a set distance. We provide detailed measurements of scat dimensions for this ape taxon which previously have been lacking. This research builds upon prior work by recording more in-depth data for focal subjects and travelling parties on defaecation and scat encounter rates. The findings presented should assist in the interpretation of scat data when censusing unhabituated chimpanzees.