Abstract
ABSTRACT Introduction: Advances in the development of nucleic acid-based methods have dramatically facilitated studies of host–microbial interactions. Fecal DNA analysis can provide information about the host’s microbiota and gastrointestinal pathogen burden. Numerous studies have been conducted in mammals, yet birds are less well studied. Avian fecal DNA extraction has proved challenging, partly due to the mixture of fecal and urinary excretions and the deficiency of optimized protocols. This study presents an evaluation of the performance in avian fecal DNA extraction of six commercial kits from different bird species, focusing on penguins. Material and methods: Six DNA extraction kits were first tested according to the manufacturers’ instructions using mallard feces. The kit giving the highest DNA yield was selected for further optimization and evaluation using Antarctic bird feces. Results: Penguin feces constitute a challenging sample type: most of the DNA extraction kits failed to yield acceptable amounts of DNA. The QIAamp cador Pathogen kit (Qiagen) performed the best in the initial investigation. Further optimization of the protocol resulted in good yields of high-quality DNA from seven bird species of different avian orders. Conclusion: This study presents an optimized approach to DNA extraction from challenging avian fecal samples.
Highlights
Advances in the development of nucleic acid-based methods have dramatically facilitated studies of host–microbial interactions
Six different DNA extraction kits were tested for fecal DNA extraction using feces from mallards
The complex matrix of mallard feces made DNA extraction challenging with very poor DNA yield, indicating the need of pretreatment and/or optimization, see Figure 1(a)
Summary
Advances in the development of nucleic acid-based methods have dramatically facilitated studies of host–microbial interactions. This study presents an evaluation of the performance in avian fecal DNA extraction of six commercial kits from different bird species, focusing on penguins. The interest in microbial ecosystems of humans and other animals has increased tremendously in recent years Many of these studies have focused on the microbiota of the gut. Due to culture-dependent analysis techniques, Levin struggled to identify gut microorganisms and concluded that the gut of most Arctic animals was sterile. In retrospect, this can be viewed as an example highlighting the crucial importance of using appropriate sample storage and analysis techniques to come to the most accurate conclusion
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
