IDENTIFICATION OF CARNIVORE FECES 961 Identification of carnivore feces by local peoples and molecular analyses Gerald L. Zuercher,Philip S. Gipson, and GeorgeC. Stewart Abstract Twoemerging research techniques offer alternatives to historicallyinvasive and expensive carnivorestudies: molecular analysis and use of traditionalecological knowledge (TEK) of indigenousand local peoples. We used a molecularanalysis of the cytochrome-bgene from fecal DNA to validate identificationof mammaliancarnivore feces collected by indigenous people and local people on the Reserva Naturaldel Bosque Mbaracayu (RNBM)in easternParaguay. At least 16 mammaliancarnivore species, severalof which are endangered, are known to occur within RNBM. We found a 100% agreement between identificationdetermined through the molecular analysis and identifications providedby indigenousand local peoples. Our resultssuggest that indigenousand local knowledge may help advance scientific knowledge about natural resources. By validatingTEK through molecular techniques, we show that both indigenousand local people can contributecredibly to studiesof the naturalresources on which they depend. Key words canids, carnivores,cytochrome-b, feces, felids, molecularanalyses, scat, traditionaleco- logical knowledge Mammalian carnivores present unique study and with their environment." Several qualities of challenges to researchers. First, carnivores often TEK have led to resistance of its acceptance by sci- are elusive and sparsely distributed, requiring large entists, including that TEK is primarily qualitative, investments of time and money to adequately sam- heavily dependent on intuition, largely holistic, and ple a given population. Second, carnivores are by spiritual (Berkes 1993). Despite these differences, nature dangerous to handle. Third, traditional study the International Union for the Conservation of methods involve capture, restraint, and tissue sam- Nature (1986) predicted that new scientific knowl- pling. Animal handling increases risks to both edge could be gained from perceptive studies of researchers and carnivores and may influence the traditional environmental knowledge systems. Use behavior of study animals. Two emerging research of TEK is increasing, as documented in a special techniques offer alternatives to more traditional section of Ecological Applications (October 2000, invasive and expensive carnivore studies: the use of Volume 10, Number 5). traditional ecological knowledge (TEK) of indige- Involvement of indigenous and local peoples can nous and local peoples and molecular analyses. be particularly important in studies of endangered While there is no single definition of TEK, a syn- or elusive species or disappearing habitats where thesis of definitions has been proposed (Berkes conservation is a goal. Indigenous and local peo- 1993:3): "TEK is a cumulative body of knowledge ples may be directly affected by changes in man- and beliefs, handed down through generations by agement of threatened habitats since often they cultural transmission, about the relationship of liv- depend on the land for their subsistence. Involving ing things (including humans) with one another them helps bridge the gap between western scien- Addressfor GeraldL. Zuercherand PhilipS. Gipson:Kansas Cooperative Fish and WildlifeResearch Unit, UnitedStates Geolog- ical Survey,Division of Biology,Kansas State University,205 LeasureHall, Manhattan,KS 66506, USA;e-mail for Zuercher: [email protected] address for Zuercher:Department of Naturaland AppliedSciences, Universityof Dubuque,2000 UniversityAvenue, Dubuque, IA 52001. Addressfor GeorgeC. Stewart:Department of DiagnosticMedicine and Pathobiology, College of VeterinaryMedicine, Kansas State University, Manhattan, KS 66506, USA. WildlifeSociety Bulletin2003, 31(4):961-970 Peer refereed 962 Wildlife Society Bulletin 2003, 31(4):961-970 tists and indigenous and local peoples, which is a common frustration (Byers 1999). When indigenous people work in a collaborative manner with western scientists, they may better under- stand the goals and out- comes of research and monitoring programs in their communities, thus enhancing the long-term relationship and improv- ing the chances of reach- ing mutually beneficial outcomes (Byers 1999). The Ache of the Mbara- cayu Reserve in eastern onca)feces on a in the Reserva Natural del ParaguayParaguay were studied Jaguar(Panthera ; path Bosque Mbaracay6, Paraguay, August1999. A pockettool I measuring 11 cm is next to the feces as a reference for size. (Hawkes et al. 1982, Hill et al. 1984, Hill and Hurtado 1996) and subsequently incorporated into moni- a molecular diagnostic technique for identifying toring and research programs (Hill et al. 1997, canid and felid feces collected within Mbaracayu 2000). The TEK of the Ache is considered central Reserve, and 2) evaluate the skill of Ache and local to management of the Mbaracayu Reserve and sur- rangers in identifying feces by comparing results of rounding buffer zone (Fundaci6n Moises Bertoni their visual field identifications with our molecular 1997). The Ache also participate in planning for identifications of the same feces. future management. They presently collect most field data for human dimensions research in the Methods reserve (Hill et al. 1997, 2000). The experiences and perceptions that Ache and local people have Study site about the forest and wildlife are extensive, but the Reserva Natural del Bosque Mbaracayui (RNBM; level of credibility associated with their beliefs is Mbaracayui Forest Nature Reserve) was a 64,400- unknown. One way to assess the accuracy and pre- hectare reserve located within the Department of cision of information provided by indigenous and Canideyu in eastern Paraguay (Figure 1). It was local peoples is to compare their reports to results managed by the Fundaci6n Moises Bertoni, a con- from modern analytical methods including molec- servation organization in Paraguay. The reserve was ular scatology. located at approximately 55? west and 24? south, Molecular scatology (term coined by Reed et al. with elevations between 150-450 m above sea 1997) helps to eliminate subjectivity in identifica- level. Rainfall was unpredictable but averaged tion of feces. The method is based on identification approximately 1,800 mm per year. Winter in east- of feces to the species level by analysis of host DNA ern Paraguay was typically dry and lasted from May (Foran et al. 1997, Paxinos et al. 1997, Hansen and to September (Sanchez 1973). Marked temperature Jacobsen 1999) from sloughed colonic epithelial fluctuations occurred seasonally, with average daily cells (Albaugh et al. 1992). The conserved regions high-low temperatures of 14-25?C in July and of the mitochondrial cytochrome-b and D-Loop 22-34?C in January. The reserve protects the regions are particularly useful to discriminate largest remaining fragment of Interior Atlantic between closely related mammalian carnivores Forest in Paraguay. It is a habitat mosaic of 19 nat- (i.e., within sympatric Canidae: Paxinos et al. 1997, ural communities and is home to at least 16 species Pilgrim et al. 1998). of mammalian carnivores (Fundaci6n Moises There were 2 objectives for our study: 1) develop Bertoni 1997). Identificationof carnivorefeces * Zuercheret al. 963 km _0' i:' F' S5 Figure 1. Location of Reserva Natural del Bosque Mbaracayu (RNBM) in eastern Paraguay,where we studied carnivore scat iden- tification in 1999-2001. Map of Paraguayis courtesy of Fundaci6n Moises Bertoni. Sites 1-7 refer to ranger stations located with RNBM: 1) Jejui'mi-reserve headquarters, 2) Lagunita, 3) Horqueta'mi biological station, 4) Aguara Nu guard station, 5) INanduro- cai guard station, 6) Carapa guard station, 7) La Morena guard station. Mammalian carnivore scats were collected from sites 1-7 within the reserve. Site 8 was the location of the village where the Ach6 collaborators live. Collection of mammalian carnivore scats same code as written on the collection bag. We We collected mammalian predator scats in RNBM gave a preliminary species identification for each beginning in July 1999 and continued through sample by one of the 6 different rangers during this September 2000. We conducted organized scat col- project. The most experienced ranger made a final lections during 2 field seasons (July-August 1999 examination and species identification for each and July-August 2000), and we opportunistically sample. We stored all samples in the field inside a collected scats found at other times. Coleman? (The Coleman Company, Inc., Wichita, We accomplished scat collection by 2 methods. Kans.) cooler and surrounded samples with ice First, we selected 7 areas within RNBM (ejui'mi, packs. We took samples to the laboratory at the Lagunita, Horqueta'mi, Aguara Nu, Nandurocai, reserve headquarters within 72 hours of collection Carapa, and La Morena; Figure 1) based on the avail- and stored in a freezer at -20?C. ability of a guard station. A total of 6 rangers par- The second collection effort involved at least 10 ticipated in collecting and identifying feces. At indigenous Ache opportunistically collecting scats each site at least one reserve ranger spent several from mammalian carnivores during their hunting days walking through the forest or cerrado in and foraging trips into the reserve from the Arroyo search of game trails and other signs. We followed Bandera Reservation (Figure 1). We provided them fresh animal trails until we encountered scats. with WhirlPak? collection bags and instructed