Abstract
Edible bird’s nest (EBN) is a popular delicacy in the Asian Pacific region originating from Indonesia, Malaysia, Thailand and Vietnam, which consist of various potential medicine value in Traditional Chinese Medicine (TCM). Thailand is one of the main exporters of EBN. However, the genetic information of EBN, a key part of molecular biology, has yet to be reported in Thailand. It is necessary to explore the genetic information of EBN in Thailand based on a quick and simple method to help protect the rights and interests of consumers. This research aimed to systematically evaluate different methods of extracting EBN DNA to improve the efficiency of the analysis of cytochrome b (Cytb) and NADH dehydrogenase subunit 2 (ND2) gene sequences, the establishment of phylogenetic trees, and the genetic information of EBN in Thailand. Additionally, we aimed to develop a quick and simple method for identifying EBN from different species based on the genetic information and amplification-refractory mutation system PCR (ARMS-PCR). By comparing the four methods [cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), kit and guanidinium isothiocyanate methods] for EBN extraction, we found that the guanidinium isothiocyanate method was the optimal extraction method. Phylogenetic trees generated on the basis of Cytb and ND2 gene analyses showed that 26 samples of house EBN and 4 samples of cave EBN came from Aerodramus fuciphagus and Aerodramus maximus, respectively. In addition, to distinguish different samples from different species of Apodiformes, we designed 4 polymerase chain reaction (PCR) amplification primers based on the ND2 gene sequences of A. fuciphagus and A. maximus. The ARMS-PCR results showed band lengths for A. fuciphagus EBN of 533, 402, and 201 bp, while those for A. maximus EBN were 463, 317, and 201 bp. Collectively, the results showed that ARMS-PCR is a fast and simple method for the genetic identification of EBN based on designing specific original identification primers.
Highlights
IntroductionEdible bird’s nest (EBN), produced by swiftlets of the Aerodramus genus (mainly Aerodramus fuciphagus and Aerodramus maximus) is constructed from viscous, sticky secretions of mucin glycoprotein from a pair of sublingual glands beneath the tongue of swiftlets (Looi et al, 2017)
Edible bird’s nest (EBN), produced by swiftlets of the Aerodramus genus is constructed from viscous, sticky secretions of mucin glycoprotein from a pair of sublingual glands beneath the tongue of swiftlets (Looi et al, 2017)
The results obtained from the four methods showed that all EBN DNA samples obtained via the guanidine isothiocyanate method were amplified successfully with a clear single band (Figure 1)
Summary
Edible bird’s nest (EBN), produced by swiftlets of the Aerodramus genus (mainly Aerodramus fuciphagus and Aerodramus maximus) is constructed from viscous, sticky secretions of mucin glycoprotein from a pair of sublingual glands beneath the tongue of swiftlets (Looi et al, 2017). EBN has been esteemed as a nutritious food since the Yuan dynasty in China, which consist of various potential medicine value in Traditional Chinese Medicine (TCM). It is widely distributed in Indonesia, Malaysia, Thailand, Huaiji, and other Southeast Asian regions along the Pacific. There are two classification of EBN: house nests (found in swiftlet houses) and cave nests (found in natural caves) (Liu et al, 2020). The EBN market value is estimated to range from $1000 to $50 000 per kg according to the swiftlet species (Liu et al, 2020). Some unethical suppliers introduce cheaper adulterants into EBN, including high-protein and high-carbohydrate substances (Huang et al, 2019)
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