The identification of species primordium has been one of the hot issues in the identification of traditional Chinese medicine. Sea snake is one of the most valuable Chinese medicinal materials in China. In order to understand the origin and varieties of sea snake in the market, we studied the molecular identification of 46 sea snakes by cytochrome B(Cytb). After comparison and manual correction, the sequence length was 582 bp, and the content of A+T(58.9%) was higher than that of G+C(41.1%). There exist 197 variable sites and 179 parsimony-informative sites of the sequence. There are 44 kinds of sequence alignment with consistency equal to 100%, and 2 kinds equal to 96%. A total of 408 Cytb effective sequences were downloaded from GenBank database, with a total of 68 species. Phylogenetic tree of a total of 454 sea snake sequences with the samples in this study were constructed by neighbor-joining trees and Bayesian inference method, respectively, which can identify 42 samples of medicinal materials, while 4 samples can not be identified because of their low node support. The results showed that the species of the sea snake medicine were at least from 2 genera and 5 species, namely, Aipysurus eydouxii, Hydrophis curtus, H. caerulescen, H. curtus, H. ornatus and H. spiralis. This study suggested that the original species of commercial sea snake are very complex and can provide insight into the identification of sea snakes.
Animals ; Bayes Theorem ; China ; Cytochromes b/genetics* ; Hydrophiidae ; Medicine, Chinese Traditional ; Phylogeny

Taenia pisiformis is one of the most important parasites of canines and rabbits. T. pisiformis cysticercus (the larval stage) causes severe damage to rabbit breeding, which results in huge economic losses. In this study, the genetic variation of T. pisiformis was determined in Sichuan Province, China. Fragments of the mitochondrial cytochrome b (cytb) (922 bp) gene were amplified in 53 isolates from 8 regions of T. pisiformis. Overall, 12 haplotypes were found in these 53 cytb sequences. Molecular genetic variations showed 98.4% genetic variation derived from intra-region. F(ST) and Nm values suggested that 53 isolates were not genetically differentiated and had low levels of genetic diversity. Neutrality indices of the cytb sequences showed the evolution of T. pisiformis followed a neutral mode. Phylogenetic analysis revealed no correlation between phylogeny and geographic distribution. These findings indicate that 53 isolates of T. pisiformis keep a low genetic variation, which provide useful knowledge for monitoring changes in parasite populations for future control strategies.
Animals ; China ; Cytochromes b/*genetics ; *Genetic Variation ; Helminth Proteins/*genetics ; Humans ; Mitochondria/*genetics ; Molecular Sequence Data ; Phylogeny ; Rabbits ; Taenia/classification/genetics/*isolation & purification ; Taeniasis/*parasitology

We report here a human case of Taenia asiatica infection which was confirmed by genetic analyses in Dali, China. A patient was found to have symptoms of taeniasis with discharge of tapeworm proglottids. By sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene, we observed nucleotide sequence identity of 99% with T. asiatica and 96% with T. saginata. Using the cytochrome b (cytb) gene, 99% identity with T. asiatica and 96% identity with T. saginata were found. Our findings suggest that taeniasis of people in Dali, China may be mainly caused by T. asiatica.
Adult ; Animals ; China ; Cytochromes b/genetics ; Electron Transport Complex IV/genetics ; Humans ; Male ; Phylogeny ; Sequence Homology, Nucleic Acid ; Taenia/classification/genetics/isolation & purification/*physiology ; Taeniasis/*parasitology

The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.
Animals ; Bungarus ; classification ; genetics ; Cytochromes b ; genetics ; DNA Primers ; genetics ; Drug Contamination ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Alignment ; Sequence Analysis, DNA

Hydrophidae, one of the precious traditional Chinese medicines, is generally drily preserved to prevent corruption, but it is hard to identify the species of Hydrophidae through the appearance because of the change due to the drying process. The identification through analysis on gene barcode, a new technique in species identification, can avoid the problem. The gene barcodes of the 6 species of Hydrophidae like Lapemis hardwickii were aquired through DNA extraction and gene sequencing. These barcodes were then in sequence alignment and test the identification efficency by BLAST. Our results revealed that the barcode sequences performed high identification efficiency, and had obvious difference between intra- and inter-species. These all indicated that Cyt b DNA barcoding can confirm the Hydrophidae identification.
Animals ; Base Sequence ; China ; Cytochromes b ; genetics ; DNA Barcoding, Taxonomic ; Elapidae ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Reptilian Proteins ; genetics ; Sequence Analysis, DNA

This study was conducted to define the taxonomic status of Spermophilus in the plague area of Dingbian County in Shaanxi Province, China, through the two-factor variance analysis of morphological characteristics, DNA barcoding, and chromosome karyotype analysis. The Spermophilus samples collected from Dingbian and Zhengxiang Baiqi Counties exhibited significant differences in their morphological measurements. All Spermophilus samples form two distinct branches in neighbor-joining (NJ) tree. One branch included the Spermophilus samples collected from Inner Mongolia, and the other branch included samples collected from the plague foci of Shaanxi Province and the Ningxia Region. The Spermophilus samples collected from Dingbian County had a chromosome number of 2n = 38 in 84.40% of all their cells. The Spermophilus species collected from the plague area of Dingbian County was categorized as Spermophilus alashanicus (S.alashamicus). The findings reported in this study are epidemiologically significant for monitoring plague in this region of west-central China.
Animals ; China ; Cytochromes b ; analysis ; DNA Barcoding, Taxonomic ; Electron Transport Complex IV ; analysis ; Karyotype ; Plague ; microbiology ; Sciuridae ; anatomy & histology ; classification ; genetics

<b>OBJECTIVEb>To explore the correlation of the mutation of MTCYB and MTATP6 genes in sperm mitochondria with asthenospermia.

<b>METHODSb>We extracted mtDNA from 80 semen samples of asthenospermia and 20 of normal sperm motility, amplified the MTCYB and MTATP6 genes by PCR, and analyzed their mutation by sequencing and BLAST matching.

<b>RESULTSb>The deletion of both MTCYB and MTATP6 were detected in 20 of the 80 asthenospermia samples, MTCYB deletion in 16 and MTATP6 deletion in 4, accounting for 20% and 5% respectively. Sequencing and BLAST matching revealed G8887A mutation in the MTATP6 gene in the asthenospermia samples, with a mutation rate of 20%, while no regular mutation was noted in MTCYB. Neither significant deletion nor mutation was observed in any of the 20 samples of normal sperm motility.

<b>CONCLUSIONb>Both the deletion and mutation of MTCYB and MTATP6 genes in sperm mitochondria might affect sperm motility in adults.

Adult ; Asthenozoospermia ; genetics ; pathology ; Base Sequence ; Cytochromes b ; genetics ; DNA, Mitochondrial ; genetics ; Humans ; Male ; Mitochondrial Proteins ; genetics ; Mitochondrial Proton-Translocating ATPases ; genetics ; Molecular Sequence Data ; Mutation ; Sequence Homology, Nucleic Acid ; Sperm Count ; Spermatozoa ; metabolism ; pathology

<b>OBJECTIVEb>To develop a convenient and effective method for the identification of Bungarus multicinctus.

<b>METHODb>Based on the sequence of Cyt b gene fragment of B. multicinctus and its adulterants, a pair of highly specific primer (HJL- and HJH-) were designed for distinguishing B. ulticinctus from other species of snake. To establish specific PCR reaction condition, the primers were employed to amplify the DNA templates extracted from B. multicinctus and 6 other species of snake, under different annealing temperature. Using this method, B. multicinctus was identified from 18 samples bought from many drugstores.

<b>RESULTb>A 230 bp DNA fragment was amplified from B. multicinctus in PCR with annealed temperature at 67 degrees C, whereas no DNA fragment was amplified from other snake samples under the same reaction condition, B. multicinctus could be clearly distinguished from others by PCR reaction with the highly specific primers. In the present study, 18 sample, bought from different drugstores, were also identified by the highly specific PCR with the primers. The results indicated that 14 samples were B. multicinctus and the other 4 were adulterant, which was consistent with the conclusion of authentication based on morphological.

<b>CONCLUSIONb>The primers designed in the present study were highly specific for B. multicinctus.

Animals ; Base Sequence ; Bungarus ; classification ; genetics ; Cytochromes b ; genetics ; DNA ; genetics ; DNA Primers ; Drug Contamination ; Materia Medica ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; Snakes ; classification ; genetics ; Species Specificity

OBJECTIVE: To establish a fluorescent multiple amplification system of 16S rRNA and Cytb genes located in mitochondrial DNA for species identification. METHODS: A pair of primers of 16S rRNA gene and Cytb gene of the mitochondrial DNA was designed with the software Primer 5.0 to construct a multiple amplification system. The amplified products from human and five species of animals, including cattle, pig, dog, chicken and grass carp were analyzed by 310 Genetic Analyzer. RESULTS: The amplified products of these samples showed two peaks. The common one was 358bp and the specific one different in unique species was between 231bp and 256bp. CONCLUSION The multiplex amplification system can exactly distinguish the species of human from five common animals.
Animals ; Base Sequence ; Carps ; Cattle ; Chickens ; Cytochromes b/genetics* ; DNA, Mitochondrial/genetics* ; Dogs ; Female ; Forensic Genetics ; Gene Amplification ; Humans ; Male ; Molecular Sequence Data ; RNA, Ribosomal, 16S/genetics* ; Species Specificity ; Swine

We designed the specific primers and TaqMan probes targeting cytochrome b genes of mitochondrial DNA from bovine, goat and sheep. We used different fluorescents to label the probes. After optimization of reaction conditions, we set up a multiplex fluorescent real-time PCR method to detect bovine, goat and sheep derived materials, simultaneously. We finished the detection tests of 17 kinds of animal DNA and 200 DNA samples from different sources with the developed method and the National Standard GB/T 20190Y-2006 routine PCR method. The coincidence rate of these two methods was 100%. Without electrophoresis or restriction digestion, the developed method could reduce the test time to one third as routine PCR and identify three kinds of animal derived materials including bovine, goat and sheep in one reaction. The developed method was approximately 10 times more sensitive than routine PCR, and was applicable to identifications of bovine, goat and sheep derived materials in feed stuff, meat, milk, pelt and grease, etc. The study showed that the developed real-time PCR method is a rapid, sensitive and efficacious detection assay for bovine, goat and sheep derived materials in animal products.
Animal Feed ; analysis ; Animals ; Cattle ; Cytochromes b ; genetics ; DNA Primers ; DNA, Mitochondrial ; analysis ; genetics ; Food Contamination ; analysis ; Genes, Mitochondrial ; Goats ; Meat ; analysis ; Mitochondria, Muscle ; enzymology ; genetics ; Polymerase Chain Reaction ; methods ; Sheep