Sex Determination of Sika Deer (Cervus nippon yesoensis) Using Nested PCR from Feces Collected in the Field

Yamazaki, Shoki; Motoi, Yuta; Nagai, Kazuya; Ishinazaka, Tsuyoshi; Asano, Makoto; Suzuki, Masatsugu

doi:10.1292/jvms.11-0235

Cited by 8 publications

(8 citation statements)

References 31 publications

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“…Our accuracy rate of sex determination was 80.2% of samples over 2 trials, which is comparable to the success rate observed for the SRY primer (85%) (Yamazaki et al, 2011). Further, the sex ratio detected here (1.27 female:1 male) corresponded to the ratio found in other field studies of blue sheep inhabiting Helan Mountain (Wang et al, 1998;Cao, 2005;Liu et al, 2007).…”

Section: Discussionsupporting

confidence: 88%

Sex identification based on AMEL gene PCR amplification from blue sheep (Pseudois nayaur) fecal DNA samples

et al. 2015

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ABSTRACT. The use of noninvasive genetic sampling to identify the sex of wild animals is an extremely valuable and important tool in molecular ecology and wildlife conservation. Sex determination using the amelogenin gene has been conducted in many species because only a single pair of primers is required to amplify both Xand Y-linked alleles. However, this method has not been used in field research with the feces of wildlife. In this study, we applied this method to 222 fecal samples from wild blue sheep (Pseudois nayaur) using amelogenin primers (SE47/SE48) after testing the effectiveness of sex determination using tissue samples and fecal samples from blue sheep of known sex. We found this method to be highly reliable (80.2%) for blue sheep. Amelogenin can be used to identify the sex of wild animals using fecal samples.

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Section: Discussionsupporting

confidence: 88%

Sex identification based on AMEL gene PCR amplification from blue sheep (Pseudois nayaur) fecal DNA samples

et al. 2015

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“…To date, DNA from fecal samples of sika deer or Japanese serow was obtained using commercial DNA extraction kits (Horino and Nagata 2004;Yamazaki et al 2011) or the DNA extraction method described by Zhang et al (2006) (Yamashiro et al 2010). However, these methods require multiple reagents, expensive equipment, and they are laborious and time-consuming.…”

Section: Dna Extraction From Fecal Pellets and Muscle Tissuesmentioning

confidence: 99%

“…However, these methods require multiple reagents, expensive equipment, and they are laborious and time-consuming. The outer layer of the fecal pellet of sika deer contains a significant amount of DNA (Yamazaki et al 2011). Accordingly, to simplify the DNA extraction from fecal pellets of both animals, we did not use commercial DNA extraction kits or the method described by Zhang et al (2006).…”

Section: Dna Extraction From Fecal Pellets and Muscle Tissuesmentioning

confidence: 99%

A novel and rapid diagnostic method for discriminating between feces of sika deer and Japanese serow by loop-mediated isothermal amplification

2015

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Severe damages to natural vegetation, agriculture, and forestry caused by overpopulation of sika deer (Cervus nippon) have markedly increased in Japan in recent years. To devise a population management plan of sika deer, information on the distribution and population size of the animal in each region is indispensable. An easy and effective method to obtain this information is to count the fecal pellets in the field. However, the habitat of sika deer in Japan overlaps that of Japanese serow (Capricornis crispus). Additionally, it is difficult to discriminate between the feces of both animals. Here, we present a rapid and precise diagnostic method for discriminating between the feces of sika deer and Japanese serow using loop-mediated isothermal amplification (LAMP) targeting cytochrome b gene in the mitochondrial DNA. Our results showed that the LAMP can discriminate between the feces of sika deer and Japanese serow, and the method is simpler and more sensitive than the conventional molecular diagnostic method. Since LAMP method does not require special skills for molecular biology techniques, even the field researchers who have never done a molecular experiment can easily carry out the protocol. In addition, the entire protocol, from DNA extraction from fecal pellet to identification of species, takes only about 75 min and does not require expensive equipment. Hence, this diagnostic method is simple, fast, and accessible to anyone. As such, the method can be a useful tool to estimate distribution and population size of sika deer.

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“…Fecal molecular biotechnology provides a rapid and dependable way of sampling endangered animals [11][12][13][14]. In addition, with the development of molecular biology technology, fecal DNA is extensively used in genetic biology studies for species identification [15][16][17], individual identification [18][19][20], sex identification [21][22][23][24][25], population genetic structure [26][27][28], and genetic diversity evaluation [29]. However, fecal sampling has some problems, such as poor fecal DNA isolation quality and low success rate of PCR amplification [30].…”

Section: Introductionmentioning

confidence: 99%

Fecal DNA Isolation and Degradation in Clam Cyclina sinensis: Noninvasive DNA Isolation for Conservation and Genetic Assessment

Zhang¹,

Wei²,

Dong

et al. 2019

Preprint

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Background To avoid destructive sampling for conservation and genetic assessment, we isolated the DNA of clam Cyclina sinensis from their feces. DNA electrophoresis and PCR amplification were used to determine the quality of fecal DNA. And we analyzed the effects of different conditions on the degradation of feces and fecal DNA.Results The clear fecal DNA bands were detected by electrophoresis, and PCR amplification using clam fecal DNA as template was effective and reliable, suggesting that clam feces can be used as an ideal material for noninvasive DNA isolation. In addition, by analyzing the effects of different environmental temperatures and soaking times on the degradation of feces and fecal DNA, we found that the optimum temperature was 4 °C. In 15 days, the feces maintained good texture, and the quality of fecal DNA was good. At 28 °C, the feces degraded in 5 days, and the quality of fecal DNA was poor.Conclusions The clam feces can be used as an ideal material for noninvasive DNA isolation. Moreover, the quality of fecal DNA is negatively correlated with environmental temperature and soaking time.

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Sex Determination of Sika Deer (Cervus nippon yesoensis) Using Nested PCR from Feces Collected in the Field

Cited by 8 publications

References 31 publications

Sex identification based on AMEL gene PCR amplification from blue sheep (Pseudois nayaur) fecal DNA samples

Sex identification based on AMEL gene PCR amplification from blue sheep (Pseudois nayaur) fecal DNA samples

A novel and rapid diagnostic method for discriminating between feces of sika deer and Japanese serow by loop-mediated isothermal amplification

Fecal DNA Isolation and Degradation in Clam Cyclina sinensis: Noninvasive DNA Isolation for Conservation and Genetic Assessment

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