A practive faeces collection protocol for multidisciplinary research in wildlife science

Abstract

24Faecal samples have become important non-invasive source of information in wildlife 25 biology and ecological research. Despite regular use of faeces, there is no universal protocol 26 available for faeces collection and storage to answer various questions in wildlife biology. 27 We collected 1408 faeces from ten different species using a dry sampling approach, and 28 achieved 94.87% and 86.02% success rate in mitochondrial and nuclear marker 29 amplifications. We also suggest a universal framework to use the same samples for different 30 use. This protocol provides an easy, quick and cheap option to collect non-invasive samples 31 from species living at different environmental conditions to answer multidisciplinary 32 questions in wildlife biology. 33 34 35 Keywords: Non-invasive wildlife research, species biology, dry sampling, variable habitat, 36 field logistics. 37 38 39 40 41 42 43 44 45 46 Non-invasive samples, in particular faeces have become a regular choice in wildlife biology, 47 population monitoring and ecological research globally. Advantages of faecal sample-based 48 wildlife research include easy sample collection, access to large sample size and spatio-49 temporal coverage. Historically, large scale use of faeces in wildlife biology started with 50 dietary analysis of animals 1 but the introduction of advanced molecular tools added a new 51 dimension to non-invasive research. These molecular tools have allowed biologists to 52 investigate questions regarding population genetics 2,3 , species distribution 4 , demography 5,6 , 53 evolutionary biology 7 and wildlife forensics 8 . In more recent time, faecal samples have been 54 used in addressing various questions related to wildlife physiology including endocrinology 55 and reproductive capacity 9,10 , along with parasitology 11,12 , disease dynamics 13 and 56 conservation genomics 14 . The sampling and storage demands of various questions in non-57 invasive wildlife research have led to a gradual development of faecal sampling and storage 58 protocols. A number of logistical factors including collector's safety, storage in the field, 59 shipping samples from remote field areas with different environmental conditions etc. have 60 been considered while gradual development of these protocols. 61Over the years, a number of faeces collection and storage approaches has been used in 62 wildlife research that are broadly classified into three categories: a) dry sampling (for 63 example simple drying 15 , silica preservation 16 ); b) wet sampling (ethanol collection 17 ; TNE 64 and DMSO buffer 18 ; DETs solution 19 ; RNA later 20 ) and c) two-step approach 21 (see Table 65 1 for details). While all of these approaches have been used in wildlife research, they have 66 several logistical limitations making their implementation in the field challenging. For 67 example, sampling with silica beads has advantages in post-collection sample transport and 68 storage 22 but is not cost effective as it requires large amount of silica beads to keep samples 6...

Standardization and validation of a panel of cross-species microsatellites to individually identify the Asiatic wild dog (Cuon alpinus)

“…not covered by dust, soil etc.) then it was swabbed with phosphate buffer saline soaked sterile cotton swab and was stored in sterile Eppendorf tube at −20 °C (Biswas et al, in press). However, if the mucous layer was covered then the top layer was scraped using sterile blades and stored in similar conditions (Biswas et al, in press).…”

“…then it was swabbed with phosphate buffer saline soaked sterile cotton swab and was stored in sterile Eppendorf tube at −20 °C (Biswas et al, in press). However, if the mucous layer was covered then the top layer was scraped using sterile blades and stored in similar conditions (Biswas et al, in press). Subsequently, faecal samples collected by both methods were lysed overnight in 300/600 μl of lysis buffer for swabs and scraped samples, respectively and 20 μl proteinase K followed by extraction using the kit’s protocol.…”

Effects of faecal inorganic contents in accurate measures of stress and nutrition hormone in large felines: implications for physiological assessments in free-ranging animals

“…Faecal samples were stored in −20 0 C freezer till further processing. To identify tiger faeces we used DNA-based approaches described in Biswas et al (2019). In brief, we swabbed each sample twice and digested the swabs overnight at 56 0 C with 30μl proteinase K and 300μl ATL buffer.…”

Peer Review #1 of "Standardization and validation of a panel of cross-species microsatellites to individually identify the Asiatic wild dog (Cuon alpinus) (v0.4)"

“…not covered by dust, soil etc.) then it was swabbed with phosphate buffer saline soaked sterile cotton swab and was stored in sterile Eppendorf tube at -20 0 C (Biswas et al, 2019). However, if the mucous layer was 2018:10:32285:4:0:NEW 7 Jul 2019) Manuscript to be reviewed covered then the top layer was scraped using sterile blades and stored in similar conditions (Biswas et al, 2019).…”

“…then it was swabbed with phosphate buffer saline soaked sterile cotton swab and was stored in sterile Eppendorf tube at -20 0 C (Biswas et al, 2019). However, if the mucous layer was 2018:10:32285:4:0:NEW 7 Jul 2019) Manuscript to be reviewed covered then the top layer was scraped using sterile blades and stored in similar conditions (Biswas et al, 2019). Subsequently, faecal samples collected by both methods were lysed overnight in 300/600 μl of lysis buffer for swabs and scraped samples, respectively and 20 μl proteinase K followed by extraction using the kit's protocol.…”