Optimization of the Cetyltrimethylammonium bromide (CTAB) DNA extraction protocol using forest elephant dung samples.

MethodsX

Laboratoire de Biotechnologie, Agriculture et Valorisation des Ressources Biologiques, Université Félix Houphouët-Boigny d'Abidjan-Cocody, Côte d'Ivoire; 22, BP 582 Abidjan, Côte d'Ivoire.

Published: September 2022

Among non-invasive biological samples, feces offer an important source of DNA and can easily be collected. However, working with fecal DNA from highly vegetarians species such as elephant is more challenging because plant secondary compounds have an inhibitory effect on PCR reactions. Working with forest elephant dung samples, we tested and adapted a protocol of DNA extraction developed on plants based on the Cetyltrimethylammonium bromide (CTAB) protocol. The protocol is relatively simple and yields a high DNA concentration. It is five-time less expensive compared to the methods of Benbouza et al. The extracted DNA is of good quality and easily amplified by PCR. The high-amplification percentage of mitochondrial genes in fecal DNA and subsequent sequencing of PCR products indicate that the proposed optimized method is reliable for molecular analysis of forest elephant dung samples.•Our optimized CTAB protocol has been adjusted by the addition of Sodium Dodecyl Sulfate (SDS) and proteinase K during the lysis phase. The combined effect of these reagents was capable of lysing cell walls and removing proteins efficiently.•Moreover, the prolonged time of incubation (overnight incubation at room temperature followed by 3 hours of incubation in a water bath) enhanced the increase of DNA yield but make the optimized protocol more time-consuming.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9558105PMC
http://dx.doi.org/10.1016/j.mex.2022.101867DOI Listing

Publication Analysis

Top Keywords

forest elephant
12
elephant dung
12
cetyltrimethylammonium bromide
8
bromide ctab
8
dna
8
dna extraction
8
dung samples
8
fecal dna
8
ctab protocol
8
protocol
6

Similar Publications

Roles of host and environment in shift of primary anthrax host species in Kruger National Park.

PLoS One

December 2024

Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa.

Environmental and climatic factors, as well as host demographics and behaviour, significantly influence the exposure of herbivorous mammalian hosts to pathogens such as Bacillus anthracis, the causative agent of anthrax. Until the early 1990s in Kruger National Park (KNP), kudu (Tragelaphus strepsiceros) was the host species most affected by anthrax, with outbreaks occurring predominantly in the dry season, particularly during drought cycles. However, the most affected host species has shifted to impala (Aepyceros melampus), with more frequent anthrax outbreaks during the wet season.

View Article and Find Full Text PDF
Article Synopsis
  • The growing human population is causing more interactions with wildlife, leading to increased conflicts, particularly with Asian elephants in the Western Ghats of India.
  • The study analyzed six years of human-elephant conflict (HEC) data alongside ecological factors to identify causes and patterns of crop damage, property damage, and human casualties.
  • Key findings indicate that HEC is influenced by habitat conditions such as grass biomass and human activities, suggesting that reducing anthropogenic pressures could alleviate conflicts.
View Article and Find Full Text PDF

Insect herbivory plays a crucial role in shaping plant communities in many terrestrial ecosystems. However, in African savannas, insect herbivory has been relatively understudied compared to large mammalian herbivory. In this study, we examined the impact of insect herbivory, focusing on leaf chewers and miners, in a South African savanna-forest mosaic (including patches of forest, thicket and savanna) in Hluhluwe iMfolozi Park, South Africa.

View Article and Find Full Text PDF

Survey-based inference of continental African elephant decline.

Proc Natl Acad Sci U S A

November 2024

Department of Fish, Wildlife, and Conservation Biology, Colorado State University, Fort Collins, CO 80523.

Long-term quantification of temporal species trends is fundamental to the assignment of conservation status, which in turn is critical for planning and targeting management interventions. However, monitoring effort and methodologies can change over the assessment period, resulting in heterogeneous data that are difficult to interpret. Here, we develop a hierarchical, random effects Bayesian model to estimate site-level trends in density of African elephants from geographically disparate survey data.

View Article and Find Full Text PDF
Article Synopsis
  • The symptoms of the disease start with small brown leaf spots that enlarge over time, and researchers collected samples to analyze the causative agent, revealing the presence of Fusarium species.
  • The study utilized molecular verification techniques, including PCR amplification, and confirmed the identity of the isolated fungi as Fusarium asiaticum, based on high sequence similarity in multiple genes across samples.
View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!