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Mini Review
Volume 6 Issue 2 - May 2018
DOI: 10.19080/JCMAH.2018.06.555683
J Complement Med Alt Healthcare
Copyright © All rights are reserved by Gallant KL Chan
Searching for Active Ingredients in Edible
Bird’s Nest
Gallant KL Chan1,2*, Kevin QY Wu1,2, Aster HY Fung2, Karmen KM Poon2, Caroline Y Wang2, Elizaveta Gridneva2,
Rena RH Huang1, Sisley YZ Fung1,2, YT Xia1,2, Winnie WH Hu1, Zack CF Wong1, 2 and Karl WK Tsim1,2
1Shenzhen Key Laboratory of Edible and Medicinal Bioresources, Shenzhen Research Institute, China
2Division of Life Science and Center for Chinese Medicine, The Hong Kong University of Science and Technology, China
Submission: April 03, 2018; Published: May 14, 2018
*Corresponding author: Gallant KL Chan, Division of Life Science and Center for Chinese Medicine, The Hong Kong University of Science and
Technology, Clear Water Bay, Hong Kong, China, Email:
J Complement Med Alt Healthcare J 6(2): JCMAH.MS.ID.555683 (2018) 001
Introduction
Edible bird’s nest (EBN; cubilose), a well-known traditional
food, used in Asia, has various nutritious and/or medicinal
      
        
secretion from the species of swiftlets, i.e. Aerodramusfuciphagus
and Aerodramus maximus 
white, yellow and red. White EBN accounts for over 90% of EBN
supply. The formation of Yellow and Red EBN are proposed to be
          
most expensive than others. According to the historical record in
AD 1700 (Qing dynasty in China), e.g. “Ben Cao Feng Yuan” and
“Ben Cao Gang Mu Shi Yi”, Red EBN possesses additional function
of replenish blood, as compared to that of White EBN. Due to the
restricted supply in the market, EBN has a high price with a range
from US$ 1,000 - 15,000 per kilogram in the food market.
N-Acetylneuraminic Acid - An Indicative Quality
Control Marker for EBN
EBN consists of 40 - 60% of protein, 20 - 30 % carbohydrate,
10 - 15% water and trace amount and minerals. Sialic acid, with
its major form as N-acetylneuraminic acid (NANA), comprises
       
of NANA present in EBN: they are either loosely attached on the
surface of EBN, namely free form of NANA, or covalently bound to
glycan molecules and linked to protein mass, namely conjugated
form of NANA. Due to the abundance of N-linked glycans present
          
form, and thus in general only a trace amount of free NANA was
detected on EBN. The amount of free form of NANA has been
proposed to be a quality control marker for authentication and
         
         
commonly used strategies for EBN authentication. However, none
of those methods were able to differentiate different grading of
EBN.
NANA was proposed to be a major component for anti-
        

study, both mushroom and human tyrosinase inhibition assay
were applied to determine the skin whitening function of
different EBN. White and Red EBN showed obvious inhibition
effect on tyrosinase activity, while the inhibition effect of Grass
        
activity for all adulterants of EBN. Moreover, NANA was able to
inhibit both mushroom and human tyrosinase activities in dose-
dependent manner, and which showed a mixed type of inhibition
         

Glycoprotein - A Longmystery on EBN
Protein comprised the largest part of EBN, over half of the nest
made of protein. However, the proteomics study was hindered
by several limitations. The heavily glycosylated chains of peptide
         

report of protein on EBN, published in 1975 by a French group,

         
Abstract
                   
Aerodramusfuciphagus), which has been consumed for several hundred years. Until now, EBN is still a popular luxurious food supplement for
women in oriental regions. According to traditional Chinese medicinal descriptions, EBN can promote the “Qi”, corresponding to the lung/
respiratory functions; and hence which improves the healthiness of skin. However, the underlying mechanism of this medicinal theory is still

How to cite this article: Gallant KL C, Kevin QY W, Aster HY F, Karmen KM P, Caroline Y W,et.al. Searching for Active Ingredients in Edible Bird’s Nest.
J Complement Med Alt Healthcare .2018; 6(2): 555683. DOI: 10.19080/JCMAH.2018.06.555683.
002
Journal of Complementary Medicine & Alternative Healthcare
     

came up after 10 years, a mitogenic factor was reported in EBN,
which expressed epithelial growth factor (EGF)-like activity via
      
       
had been achieved for the following 25 years. Since 2012, an
increasing number of reports about the proteomic study of EBN



       
extension in knowledge of protein in EBN, new authentication
methods targeting protein identities of EBN were developed as

        
        

Table 1:
No. Identified Protein Year of Published Library origin
1 2017 Chaetura pelagic
2 2017 Chaetura pelagic
3 2017 Chaetura pelagic
4 2017 Chaetura pelagic
5 2017 Chaetura pelagic
6 2017 Chaetura pelagic
7 2015 Gallus gallus
8 2015 Gallus gallus
9 2015 Gallus gallus
10  2013 Swiss-Prot
11  2001 Swiss-Prot
12  1987 Unknown
13  1977 Unknown
        
         
Trolox standards. The protein of EBN was found to be responsible
for the anti-oxidation function, which eventually led to anti-


Nevertheless, the mechanism for such peptide to execute its
function on anti-ageing was still a mystery.
Nitrite - A Hidden reat in EBN
The discovery of nitrite contamination in 2011, reported by
Chinese authority in Zhejiang province, aroused public concern
on the safety of EBN consumption. The highest amount of nitrite
on EBN (Red blood EBN) could reached 11,000ppm and resulted
in a ban on importing EBN immediately. Nitrite is widely used as a

cured meats and other food products. Nearly all processed meats
contain nitrite; however, the usage is under a strict regulation.
    
      
breathing. Syndromes of nitrite poisoning could be chronic
        

A market survey of nitrite content in EBN, supported by
Hong Kong Merchant Association of Chinese Medicine, was held
immediately after the incident. The median nitrite content of
different types of EBN was measured, which ranged from 100-
600ppm. Furthermore, under standard processing method, up
to 98% of nitrite could be removed from EBN, and subsequently
nitrite was not detected in stewed EBN. On the other hand, the
origin of nitrite contamination was still a mystery. To search the
source of nitrite, droppings from swiftlets and water samples
were collected from the production sites of EBN in Malaysia and



by mass spectroscopy, which converted nitrate to nitrite in EBN.
         
developing EBN, successfully abolished the nitrate reduction
        
content of nitrite in EBN. This phenomenon was successfully

result of the contaminating and nitrate and the microbial nitrate

How to cite this article: Gallant KL C, Kevin QY W, Aster HY F, Karmen KM P, Caroline Y W,et.al. Searching for Active Ingredients in Edible Bird’s Nest.
J Complement Med Alt Healthcare .2018; 6(2): 555683. DOI: 10.19080/JCMAH.2018.06.555683.
003
Journal of Complementary Medicine & Alternative Healthcare
Figure 1: Proposed origin and pathway for nitrite contamination onEBN.
(1): The root nodules of plant on island type habitat contain abundant nitrogen xing bacteria. Also, fertilizers applied on the crops land
contain high amount of nitrate.
(2): Nitrate and nitrogen xing bacteria accumulate in swiftlet through the intakes of contaminated ying insects.
(3): High content of nitrate has been found in swiftlet’s droppings, and nitrate reductase deriving nitrogen xing bacteria has been
identied in EBN.
(4): The nest is made of the salivary secretion of the swiftlet and embedded with droppings at the centre of the nest.
(5): Under favourable environment, nitrate reductase convert nitrate to nitrite efciently.
(6): EBN is changed from white to yellow and then red due to the accumulation of nitrite.
Although the illustration of safe consumption method for
EBN, the industry still suffers from dramatic economic loss
until 2014. The global market value of EBN was evaporated by
thirty percent after the nitrite incident in 2011, but which was
          
certain that what we achieved and described above was just a
beginning. More innovative applications of using EBN materials
will become notable in the near future.
How to cite this article: Gallant KL C, Kevin QY W, Aster HY F, Karmen KM P, Caroline Y W,et.al. Searching for Active Ingredients in Edible Bird’s Nest.
J Complement Med Alt Healthcare .2018; 6(2): 555683. DOI: 10.19080/JCMAH.2018.06.555683.
004
Journal of Complementary Medicine & Alternative Healthcare
Acknowledgement
This study was supported by Hong Kong RGC Theme-
    
     

Committee Research Grant (JCYJ 20160229205726699, JCYJ
20160229205812004, JCYJ 20160229210027564, CKFW
2016082916015476, JCYJ 20170413173747440, ZDSYS 201
707281432317 and 20170326). Z.W. received a scholarship from
HKJEBN Limited.
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How to cite this article: Gallant KL C, Kevin QY W, Aster HY F, Karmen KM P, Caroline Y W,et.al. Searching for Active Ingredients in Edible Bird’s Nest.
J Complement Med Alt Healthcare .2018; 6(2): 555683. DOI: 10.19080/JCMAH.2018.06.555683.
005
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DOI: 10.19080/JCMAH.2018.06.555683
... EBN products exported from Malaysia and Indonesia need to be cleaned in accordance with standard operating procedures to reach the EBN safety level required by China. The industry still suffers huge economic losses, and the price of bird's nest remained stable after 2016 (Chan et al., 2018). ...
... According to the report of the Chinese government in Zhejiang Province, the discovery of nitrite pollution in 2011 has aroused public concerns about the safety of EBN consumption. It also aroused the public's suspicion whether these edible EBNs are really "edible" (Paydar et al., 2013;Quek et al., 2015;Chan et al., 2018). Nitrate (NO 3 ) consists of one nitrogen atom and three oxygen atoms; while nitrite (NO 2 ) consists of one nitrogen atom and two oxygen atoms. ...
... They have successfully proved this possibility in their published study. Therefore, the nitrite on EBN may be the result of environmental pollution and nitrate and microbial nitrate reductase (Chan 2013;Chan et al., 2013;Chan et al., 2018). Swiftlet dropping/guano is source of nitrite content in EBNs, MS 2273:2012 has suggested that farmers should apply Effective Microorganism (EM) on the guano and mine the guano frequently and not allow it to build up. ...
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Edible bird’s nest (EBN) is recognized as a nourishing food among Chinese people. The efficacy of EBN was stated in the records of traditional Chinese medicine and its activities have been reported in many researches. Malaysia is the second largest exporter of EBNs in the world, after Indonesia. For many years, EBN trade to China was not regulated until August 2011, when a safety alert was triggered for the consumption of EBNs. China banned the import of EBNs from Malaysia and Indonesia due to high level of nitrite. Since then, the Malaysia government has formulated Malaysia Standards for swiftlet farming (MS 2273:2012), edible bird’s nest processing plant design and management (MS 2333:2010), and edible bird’s nest product quality (MS 2334:2011) to enable the industry to meet the specified standards for the export to China. On the other hand, Indonesia's EBN industry formulated a standard operating procedure (SOP) for exportation to China. Both countries can export EBNs to China by complying with the standards and SOPs. EBN contaminants may include but not limited to nitrite, heavy metals, excessive minerals, fungi, bacteria, and mites. The possible source of contaminants may come from the swiftlet farms and the swiftlets or introduced during processing, storage, and transportation of EBNs, or adulterants. Swiftlet house design and management, and EBN processing affect the bird’s nest color. Degradation of its optical quality has an impact on the selling price, and color changes are tied together with nitrite level. In this review, the current and future prospects of EBNs in Malaysia and Indonesia in terms of their quality, and the research on the contaminants and their effects on EBN color changes are discussed.
... In the food market, red blood EBN is the most expensive type with cost ranging from US$ 1000 to US$ 15 000 per kilogram [4]. According to historical records from 1700 (Qing dynasty in China), red blood EBN provides more health benefits than white EBN [23]. However, this special EBN has a hidden threat. ...
... In 2011, Chinese authorities reported nitrite contamination in Zhejiang Province, thus raising public concern about the safety of EBN consumption. The highest reported nitrite concentration in red blood EBN has reached 11 000 ppm, which led to the immediate ban on importing EBNs [23]. The color of cave EBN is associated with its nitrite and nitrate contents [6]. ...
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The color of edible bird's nest is associated with its nitrite concentration, but this relationship remains inconclusive. This investigation aimed to evaluate the nitrite content in edible bird's nest of four different colors: white, yellow, orange, and red blood. Fifty-eight edible bird's nest samples were obtained from five swiftlet farmhouses in Borneo Island, Indo-nesia and analyzed for nitrite content using Genesys 30 visible spectrophotometer. Results showed that the dark-colored edible bird's nests (yellow, orange, and red blood) had higher nitrite concentrations of 304, 317, and 309 ppm, respectively , compared with the white-colored one (15 ppm). Therefore, the color of edible bird's nest was associated with its nitrite concentration. This study provided updated information about the nitrite concentration in edible bird's nest of various colors.
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Edible bird's nest (EBN) swiftlet existed naturally 48,000 years ago in caves as their natural dwellings. Nowadays, edible bird's nest has become a very important industry due to its high nutritional, medicinal and economic value. Additionally, edible bird's nest has a long quality guarantee period. Obviously, the nutritional components and medicinal functions vary depending on geographical origins. Recently, the global demand for edible bird's nest has markedly increased, accompanied by the increasing attention of all key players of the global food trade system, i.e., producers, consumers, traders and the authorities to obtain safe and high-quality edible bird's nest. Hence, this target can be accomplished via the enforcement of an efficient and universal geo-tracing technique. Current methods of the geo-tracking of edible bird's nest, i.e., automation, physical and analytical techniques have several limitations and all of them fail to discriminate different quality grades of edible bird's nest. Meanwhile, in many studies and applications, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) has proven to be a “cutting edge” technique for greatly enhance food traceability from field to fork through its ability in distinguishing the food products in terms of their quality and safety. This article provides an overview of (1) edible bird's nest as a multiuse strategic food product, (2) quality issues associated with edible bird’s nest including implications that the site of acquisition of the edible bird’s nest has food safety implications, (3) current regulations and geo-tracking approaches to ensure the safety and quality of edible bird’s nest with the special focus on polymerase chain reaction-denaturing gradient gel electrophoresis technique as a vigorous and universal geo-tracing tool to be suggested for edible bird's nest geo-traceability.
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Background Edible swiftlet's nest (ESN) is dried gelatinized saliva secreted by swiftlets during the breeding season. The ESN has been widely consumed as a food and medicine since the ancient dynasty of China, particularly in the practice of Traditional Chinese Medicine (TCM). As a food with health-promoting effects, this made ESN a potential functional food. Whereby, functional food is food that can be consumed in the daily diet which then enhanced human health through nutritional aspect, but not as the cure of a disease. Scope and approach In this review, we discussed ESN as a potential functional food through the bioactive nutritional components and health benefits of ESN in promoting good health. Scientific evidence has proven that ESN consists of the unique glycoprotein of great value which provides high nutritional and functional properties for human health benefits. These include anti-ageing, anti-hypertension, immunity and neurological enhancement contributed by not only the unique glycoprotein but also sialic acid, epidermal growth factor (EGF) and other bioactive compounds. Key findings and conclusions The ESN appeared to be categorized as a functional food, with various functionality, applications, nutritional composition and therapeutic benefits committed by its components. This review also pronounces recent development of ESN as hydrolysate in the form of bioactive glycopeptide with improvements in terms of solubility, functional and nutritional benefits that broaden its applications in various industries.
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Considered a widespread gamebird found across the northern Afrotropical region, the Stone Partridge Ptilopachus petrosus is used in magical rituals and for traditional medicine in Africa. Although considered to be a species of Least Concern by the IUCN, loss of their habitat and hunting has caused them to be locally threatened. The aim of this ethno-ornithological study was to assess the utilization of Stone Partridge through surveys conducted in traditional communities. Some 384 people across 13 ethnic groups in Benin Republic were interviewed. Standardised ethnozoological indices were used to evaluate the importance of Stone Partridge as a source of food and a socio-cultural aspect in Benin per ethnic groups, gender and age class. Some 91.15% of respondents cited food as the major use category of Stone Partridge, followed by traditional medicinal and magical ritual. Responses varied by ethnic group and gender, but not age. Respondents reported that hunting is a major threat, leading to population declines. Stone Partridge plays an important role in rural communities in the study area, but a national conservation management strategy is lacking for this and other gamebird species.
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In order to reveal the color formation mechanism of blood-red edible bird’s nests (EBNs) and develop a quick and specific strategy to distinguish the artificial fake one, multiple methods of UPLC-TOF/MS, UV, NMR, FT-IR and 2D IR were used to detect the chemical markers of the reddening reaction, the results showed that the reddening substances were C9H10N2O5 and C9H9NO6, which were verified as products of a phenol-keto tautomerism evolved from L-tyrosine. Moreover, natural and artificial red EBNs with varying degrees of chemical fumigation also can be successfully distinguished using the chemical markers, and the protein variation in SDS-PAGE gel could also support the distinction. This work established a systematic method of chemical identification for both natural and artificial blood-red EBNs, and provided a new identification strategy for food safety control that can promote the development of a healthier market of EBNs.
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Edible bird’s nest (EBN) is widely consumed as a precious food and traditional medicine amongst Chinese. In present study, for the first time, the antioxidant properties of EBN pepsin-trypsin hydrolysate of swiftlet species Aerodramus fuciphagus and its ultrafiltration fractions were investigated. Thirteen peptides with molecular weights between 514.29 to 954.52 Da were identified in EBN fraction with the aim of mass spectrometry. Two novel pentapeptides Pro-Phe-His-Pro-Tyr and Leu-Leu-Gly-Asp-Pro, corresponding to f134-138 and f164-168 of cytochrome b of A.fuciphagus indicated the highest ORAC values of 14.95 and 14.32 µM of TE µM-1 peptide, respectively. Both purified peptides showed resistance against simulated gastrointestinal proteases. In addition, both peptides had no in vitro cytotoxicity in human lung MRC-5 cells and prevented human liver carcinoma HepG2 cellular damage caused by hydroxyl radicals. Therefore, it is suggested that EBN protein hydrolysates are good source of natural antioxidants and could be applied as nutraceutical compounds.
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Background Edible Bird’s Nest (EBN) as a popular traditional Chinese medicine is believed to have health enhancing and antiviral activities against influenza A virus (IAV); however, the molecular mechanism behind therapeutic effects of EBN is not well characterized. Methods In this study, EBNs that underwent different enzymatic preparation were tested against IAV infected cells. 50% cytotoxic concentration (CC50) and 50% inhibitory concentration (IC50) of the EBNs against IAV strain A/Puerto Rico/8/1934(H1N1) were determined by HA and MTT assays. Subsequently, the sialic acid content of the used EBNs were analyzed by fluorometric HPLC. Western Blotting and immunofluorescent staining were used to investigate the effects of EBNs on early endosomal trafficking and autophagy process of influenza virus. ResultsThis study showed that post inoculations of EBNs after enzymatic preparations have the highest efficacy to inhibit IAV. While CC50 of the tested EBNs ranged from 27.5–32 mg/ml, the IC50 of these compounds ranged between 2.5–4.9 mg/ml. EBNs could inhibit IAV as efficient as commercial antiviral agents, such as amantadine and oseltamivir with different mechanisms of action against IAV. The antiviral activity of these EBNs correlated with the content of N-acetyl neuraminic acid. EBNs could affect early endosomal trafficking of the virus by reducing Rab5 and RhoA GTPase proteins and also reoriented actin cytoskeleton of IAV infected cells. In addition, for the first time this study showed that EBNs can inhibit intracellular autophagy process of IAV life cycle as evidenced by reduction of LC3-II and increasing of lysosomal degradation. Conclusions The results procured in this study support the potential of EBNs as supplementary medication or alternative to antiviral agents to inhibit influenza infections. Evidently, EBNs can be a promising antiviral agent; however, these natural compounds should be screened for their metabolites prior to usage as therapeutic approach.
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Background: Edible Bird's nest (EBN) is an antioxidant-rich supplement that is popular in many parts of Asia. Its antioxidant and anti-inflammatory properties have been reported using in vitro system. This paper aimed to determine the antioxidant and anti-inflammatory effects of EBN in in high fat diet induced rats model. Methods: We evaluate if those properties can be translated in rats. High fat diet (HFD) was fed to rats for 12 weeks to determine its effects on oxidative stress and inflammation, and compared with HFD + Simvastatin and HFD + EBN (2.5 or 20 %). Weights were measured weekly, while serum and hepatic markers of oxidative stress (total antioxidant status and TBARS) and inflammation (interleukin 6 [IL-6], C-reactive protein [CRP] and tumor necrosis factor alpha [TNF-α]) were determined at the end of the intervention. In addition, transcriptional changes in hepatic antioxidant (superoxide dismutase, glutathione reductase, glutathione peroxidase) and inflammation (C-reactive protein, chemokine [C-C] motif 2, nuclear factor kappa beta 1 and tumor necrosis factor alpha) genes were evaluated. Results: The results showed increases in oxidative stress (raised TBARS and lowered total antioxidant status) and inflammatory markers (raised CRP, IL-6 and TNF-α) in HFD induced rats with corresponding attenuation of antioxidant gene expression and potentiation of inflammatory gene expression. EBN on the other hand attenuated the HFD-induced inflammation and oxidative stress and produced overall better outcomes in comparison with simvastatin. Conclusions: In aggregate, the results support the evidence-based utilization of EBN as a supplement for preventing obesity-related inflammation and oxidative stress in rats. These promising results can open up opportunities for translating the benefits of EBN to humans.
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Abbreviations: NANA: N-acetylneuraminic acid, EBN: Edible bird's nest, LC-MS/MS, QQQ: Triplequadrupoles liquid chromatography tandem mass spectrometry Abstract Edible bird nest (EBN) is a common health food consumed in Asia. The exaggerated skincare functions resulted in the abnormally high market value. Fake EBN started to appear in the market for the sake of its profits. A reliable authenticate method is in urgent need. Here, the determination of a free form of sialic acid, N-acetylneuraminic acid (NANA), was developed to distinguish EBN according to their grading. High amount of free NANA was revealed in red and yellow EBN: both white and grass EBN contained free NANA at low level. Unlike total NANA content measurement, fake EBN did not show detectable amount of free NANA. Moreover, the water extract of EBN showed anti-tyrosinase activity, and which was in line to amount of free NANA. Thus, the amount of free NANA could differentiate fake EBN from the genuine one, as well as its grading. In conclusion, the quantitation of free NANA by triplequadrupoles liquid chromatography tandem mass spectrometry (LC-MS/MS QQQ) was shown to be the best method in EBN authentication. In addition, the amount of free NANA in EBN was in accord to anti-tyrosinase activity of EBN.
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Edible bird's nest (EBN) is a glue-like substance deriving from salivary secretion by specific swiftlets, and protein is considered as the main component of EBN. Accounting over 50% by weight, the exact identities of EBN proteins are still not well understood, due to difficulties of extraction, purification and identification. By using EBN proteins as antigens, 31 monoclonal antibodies specifically against the proteins were generated. The proteins of EBN were subjected to identification by shotgun proteomics. Six protein identities were revealed, including acidic mammalian chitinase (AMCase)-like, mucin 5AC-like and ovoinhibitor-like proteins. In parallel, the monoclonal antibodies were used to immunoprecipitate proteins from EBN extract, and subsequently the precipitated product(s) was identified. AMCase-like protein was most frequently precipitated by the antibodies. The existence of AMCase-like protein in EBN was further verified by: (i) recognition of chicken AMCase by our anti-EBN antibodies; and (ii) recognition of EBN AMCase-like protein by a commercial anti-AMCase antibody. The antibody was highly sensitive and selective to AMCase-like protein in EBN products, with limit of detection at 0.01 μg/mL in ELISA test. Thus, AMCase-like protein, or its antibody, could be used as a new quality control marker for EBN.
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Porcine gelatine is a common adulterant found in edible bird's nests (EBN) used to increase the net weight prior to sale. This study aimed to develop indirect enzyme-linked immunosorbent assays (ELISAs) for porcine gelatine adulteration using anti-peptide polyclonal antibodies. Three indirect ELISAs were developed (PAB 1, 2 and 3), which had limits of detection (LODs) of 0.12, 0.10 and 0.11 µg/g, respectively. When applied to standard solutions of porcine gelatine, the inter- and intra-assays showed coefficient of variation (CVs) less than 20% and were able to detect at least 0.5 ng/µg (0.05%) porcine gelatine in spiked samples. The proposed ELISA offers attractions for quality control in the EBN industry.
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Edible bird’s nest (EBN) is a functional food constructed with swiftlets’ salivary glue. Counterfeit EBN products have been found in the market due to limited supply and high price of genuine EBN. In this article, a method for genetic identification of EBN was developed. The technique is based on sequence of cytochrome b gene in mitochondrial DNA. The sample sequences together with the sequences of swiftlets in GenBank were used to construct phylogenetic trees for genetic identification of samples. This method was applied to 11 EBN samples, one instant EBN soup product from Indonesia, and Huaiji EBN, a counterfeit EBN in some regions of China. Results showed that all the EBN samples and the instant EBN soup were from Aerodramus fuciphagus while the Huaiji EBN sample was from Apus nipalensis. This was consistent with identification based on morphology of the samples. Therefore, this method is a promising tool to identify the species of bird producing a given sample of EBN, and thus could be used to authenticate—that is, distinguish authentic from counterfeit—EBN.