1.Prevalence of Anaplasma and Bartonella spp. in Ticks Collected from Korean Water Deer (Hydropotes inermis argyropus).
Jun Gu KANG ; Sungjin KO ; Heung Chul KIM ; Sung Tae CHONG ; Terry A KLEIN ; Jeong Byoung CHAE ; Yong Sun JO ; Kyoung Seong CHOI ; Do Hyeon YU ; Bae Keun PARK ; Jinho PARK ; Joon Seok CHAE
The Korean Journal of Parasitology 2016;54(1):87-91
Deer serve as reservoirs of tick-borne pathogens that impact on medical and veterinary health worldwide. In the Republic of Korea, the population of Korean water deer (KWD, Hydropotes inermis argyropus) has greatly increased from 1982 to 2011, in part, as a result of reforestation programs established following the Korean War when much of the land was barren of trees. Eighty seven Haemaphysalis flava, 228 Haemaphysalis longicornis, 8 Ixodes nipponensis, and 40 Ixodes persulcatus (21 larvae, 114 nymphs, and 228 adults) were collected from 27 out of 70 KWD. A total of 89/363 ticks (266 pools, 24.5% minimum infection rate) and 5 (1.4%) fed ticks were positive for Anaplasma phagocytophilum using nested PCR targeting the 16S rRNA and groEL genes, respectively. The 16S rRNA gene fragment sequences of 88/89 (98.9%) of positive samples for A. phagocytophilum corresponded to previously described gene sequences from KWD spleen tissues. The 16S rRNA gene fragment sequences of 20/363 (5.5%) of the ticks were positive for A. bovis and were identical to previously reported sequences. Using the ITS specific nested PCR, 11/363 (3.0%) of the ticks were positive for Bartonella spp. This is the first report of Anaplasma and Bartonella spp. detected in ticks collected from KWD, suggesting that ticks are vectors of Anaplasma and Bartonella spp. between reservoir hosts in natural surroundings.
Anaplasma/genetics/*physiology
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Animals
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Arachnid Vectors/microbiology
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Bartonella/genetics/*physiology
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Chaperonin 60/genetics
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Deer/parasitology
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Disease Reservoirs/veterinary
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RNA, Ribosomal, 16S/genetics
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Republic of Korea/epidemiology
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Ticks/*microbiology
2.Genetic Diversity of Toxoplasma gondii Strains from Different Hosts and Geographical Regions by Sequence Analysis of GRA20 Gene.
Hong Rui NING ; Si Yang HUANG ; Jin Lei WANG ; Qian Ming XU ; Xing Quan ZHU
The Korean Journal of Parasitology 2015;53(3):345-348
Toxoplasma gondii is a eukaryotic parasite of the phylum Apicomplexa, which infects all warm-blood animals, including humans. In the present study, we examined sequence variation in dense granule 20 (GRA20) genes among T. gondii isolates collected from different hosts and geographical regions worldwide. The complete GRA20 genes were amplified from 16 T. gondii isolates using PCR, sequence were analyzed, and phylogenetic reconstruction was analyzed by maximum parsimony (MP) and maximum likelihood (ML) methods. The results showed that the complete GRA20 gene sequence was 1,586 bp in length among all the isolates used in this study, and the sequence variations in nucleotides were 0-7.9% among all strains. However, removing the type III strains (CTG, VEG), the sequence variations became very low, only 0-0.7%. These results indicated that the GRA20 sequence in type III was more divergence. Phylogenetic analysis of GRA20 sequences using MP and ML methods can differentiate 2 major clonal lineage types (type I and type III) into their respective clusters, indicating the GRA20 gene may represent a novel genetic marker for intraspecific phylogenetic analyses of T. gondii.
Animals
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Base Sequence
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Brazil
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China
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Deer
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*Genetic Variation
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Genotype
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Goats
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Humans
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Molecular Sequence Data
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Phylogeny
;
Protozoan Proteins/*genetics/metabolism
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Sheep
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Swine
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Toxoplasma/classification/*genetics/isolation & purification/parasitology/physiology
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Toxoplasmosis/*parasitology
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Toxoplasmosis, Animal/*parasitology
;
United States
3.Sequence Diversity in MIC6 Gene among Toxoplasma gondii Isolates from Different Hosts and Geographical Locations.
Zhong Yuan LI ; Hui Qun SONG ; Jia CHEN ; Xing Quan ZHU
The Korean Journal of Parasitology 2015;53(3):341-344
Toxoplasma gondii is an opportunistic protozoan parasite that can infect almost all warm-blooded animals including humans with a worldwide distribution. Micronemes play an important role in invasion process of T. gondii, associated with the attachment, motility, and host cell recognition. In this research, sequence diversity in microneme protein 6 (MIC6) gene among 16 T. gondii isolates from different hosts and geographical regions and 1 reference strain was examined. The results showed that the sequence of all the examined T. gondii strains was 1,050 bp in length, and their A + T content was between 45.7% and 46.1%. Sequence analysis presented 33 nucleotide mutation positions (0-1.1%), resulting in 23 amino acid substitutions (0-2.3%) aligned with T. gondii RH strain. Moreover, T. gondii strains representing the 3 classical genotypes (Type I, II, and III) were separated into different clusters based on the locus of MIC6 using phylogenetic analyses by Bayesian inference (BI), maximum parsimony (MP), and maximum likelihood (ML), but T. gondii strains belonging to ToxoDB #9 were separated into different clusters. Our results suggested that MIC6 gene is not a suitable marker for T. gondii population genetic studies.
Amino Acid Sequence
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Animals
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Base Sequence
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Cats
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Cell Adhesion Molecules/chemistry/*genetics/metabolism
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Deer
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*Genetic Variation
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Genotype
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Goats
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Humans
;
Molecular Sequence Data
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Phylogeny
;
Protozoan Proteins/chemistry/*genetics/metabolism
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Sequence Alignment
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Sheep
;
Swine
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Toxoplasma/classification/*genetics/isolation & purification/physiology
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Toxoplasmosis/*parasitology
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Toxoplasmosis, Animal/*parasitology
4.Research progress on molecular genetics of forest musk deer.
Hang JIE ; Cheng-li ZHENG ; Jian-ming WANG ; Xiao-lan FENG ; De-jun ZENG ; Gui-jun ZHAO
China Journal of Chinese Materia Medica 2015;40(22):4319-4323
Forest musk deer is one of the large-scale farming musk deer animals with the largest population at the same time. The male musk deer can secrete valuable medicines, which has high medicinal and economic value. Due to the loss of habitat and indiscriminate hunting, the numbers of wild population specie and the distribution have been drastically reduced. Therefore, in-depth understanding of the molecular genetics progress of forest musk deer will pave a way for musk deer protection and breeding. In this review, the progress associated with the molecular marker, genetic classification, artificial breeding, musk secretion and disease in past decades were reviewed, in order to provide a theoretical basis for subsequent molecular genetic researches in forest musk deer.
Animals
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Breeding
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Deer
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classification
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genetics
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growth & development
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metabolism
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Ecosystem
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Fatty Acids, Monounsaturated
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chemistry
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metabolism
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Female
;
Male
5.Molecular identification of hairy antler by analysis of high resolution melting.
Kang CHEN ; Chao JIANG ; Yuan YUAN ; Lu-qi HUANG ; Yan JIN
China Journal of Chinese Materia Medica 2015;40(4):619-623
High resolution melting (HRM) , an important technology for genotyping and mutation scanning, has broad prospects in the authenticity of traditional Chinese medicine. This paper selected universal CO I primers and used HRM to establish a new method for authenticity of Hairy Antler. PCR was conducted at the annealing temperature of 60 °C and 45 cycles. The range of the DNA template concentration, the primer concentration and the Mg2+ ion concentration were further optimized. The results showed that the Tm values of Cervus nippon were (81.96 ± 0.07), (84.51 ± 0.03) °C and Cervus elaphus was(82.58 ± 0.13), (85.95 ± 0.05) °C with 10-100 mg · L(-1) DNA template, 0.2 µLmol · L(-1) primer, 2.0 mmol · L(-1) Mg2+. This method can authenticate of hairy antler and is simple, fast, high-throughput, visualization.
Animals
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Antlers
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chemistry
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DNA
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chemistry
;
genetics
;
DNA Primers
;
genetics
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Deer
;
classification
;
genetics
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Genotype
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Medicine, Chinese Traditional
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standards
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Polymerase Chain Reaction
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Transition Temperature
6.Quality evaluation on different specifications of cervi cornu pantotrichum with its effect on ovariectomized osteoporosis model rats.
China Journal of Chinese Materia Medica 2014;39(12):2326-2329
OBJECTIVETo study the effect of eight specifications of Cervi Cornu Pantotrichum on the osteoporosis of ovariectomized rats and grade the eight different specifications of Cervi Cornu Pantotrichum.
METHODTotally 100 SD female rats were divided randomly into 10 groups, namely the normal group, the model group and eight Cervi Cornu Pantotrichum groups of different specifications. Their bilateral ovaries were excised to reproduce the osteoporosis model. Meanwhile, the rats were given the eight different specifications of Cervi Cornu Pantotrichum for consecutively 12 weeks. Subsequently, the effects of the different specifications of Cervi Cornu Pantotrichum on bone mineral density and serum biochemical indicators of rats were observed. A clustering analysis was made for the eight specifications of Cervi Cornu Pantotrichum, with the serum content of ALP, BMP-2 and BGP as influencing factors.
RESULTAfter 12 weeks, the eight different specifications of Cervi Cornu Pantotrichum could significantly improve ALP, BMP-2, BGP in serum and bone mineral density of ovariectomized rats. And the cluster analysis showed similar results to the quality classification of traditional commercial herbs Cervi Cornu Pantotrichum.
CONCLUSIONDifferent specifications of Cervi Cornu Pantotrichum can antagonize the osteoporosis of ovariectomized rats, and their effects are related to the quality of commercial herbs.
Animals ; Bone Density ; drug effects ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Deer ; Disease Models, Animal ; Female ; Horns ; chemistry ; Humans ; Osteoporosis, Postmenopausal ; drug therapy ; genetics ; metabolism ; physiopathology ; Ovariectomy ; Rats ; Rats, Sprague-Dawley
7.Genotype-4 hepatitis E in a human after ingesting roe deer meat in South Korea.
Ja Yoon CHOI ; Jeong Mi LEE ; Yun Won JO ; Hyun Ju MIN ; Hyun Jin KIM ; Woon Tae JUNG ; Ok Jae LEE ; Haesun YUN ; Yeong Sil YOON
Clinical and Molecular Hepatology 2013;19(3):309-314
The recent increase in the number of cases of indigenous hepatitis E virus (HEV) infection highlights the importance of identifying the transmission routes for the prevention of such infections. Presented herein is the first case of acute HEV infection after ingesting wild roe deer meat in South Korea. A 43-year-old male presented with abdominal discomfort and jaundice. He had not recently traveled abroad, but had eaten raw roe-deer meat 6-8 weeks before the presentation. On the 7th day of hospitalization the patient was diagnosed with acute viral hepatitis E. Phylogenetic analysis of his serum revealed genotype-4 HEV. This case supports the possibility of zoonotic transmission of HEV because the patient appears to have been infected with genotype-4 HEV after ingesting raw deer meat.
Adult
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Alanine Transaminase/blood
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Animals
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Bilirubin/blood
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Deer/virology
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Genotype
;
Hepatitis E/*diagnosis/transmission/virology
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Hepatitis E virus/classification/*genetics/isolation & purification
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Humans
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Male
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Phylogeny
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RNA, Viral/analysis
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Republic of Korea
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Travel
8.Sequencing and analysis of complete genome of rabies viruses isolated from Chinese Ferret-Badger and dog in Zhejiang province.
Yong-Liang LEI ; Xiao-Guang WANG ; Xiao-Yan TAO ; Hao LI ; Sheng-Li MENG ; Xiu-Ying CHEN ; Fu-Ming LIU ; Bi-Feng YE ; Qing TANG
Chinese Journal of Virology 2010;26(1):45-52
Based on sequencing the full-length genomes of four Chinese Ferret-Badger and dog, we analyze the properties of rabies viruses genetic variation in molecular level, get the information about rabies viruses prevalence and variation in Zhejiang, and enrich the genome database of rabies viruses street strains isolated from China. Rabies viruses in suckling mice were isolated, overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses from Chinese Ferret-Badger, dog, sika deer, vole, used vaccine strain were determined. The four full-length genomes were sequenced completely and had the same genetic structure with the length of 11, 923 nts or 11, 925 nts including 58 nts-Leader, 1353 nts-NP, 894 nts-PP, 609 nts-MP, 1575 nts-GP, 6386 nts-LP, and 2, 5, 5 nts- intergenic regions(IGRs), 423 nts-Pseudogene-like sequence (psi), 70 nts-Trailer. The four full-length genomes were in accordance with the properties of Rhabdoviridae Lyssa virus by BLAST and multi-sequence alignment. The nucleotide and amino acid sequences among Chinese strains had the highest similarity, especially among animals of the same species. Of the four full-length genomes, the similarity in amino acid level was dramatically higher than that in nucleotide level, so the nucleotide mutations happened in these four genomes were most synonymous mutations. Compared with the reference rabies viruses, the lengths of the five protein coding regions had no change, no recombination, only with a few point mutations. It was evident that the five proteins appeared to be stable. The variation sites and types of the four genomes were similar to the reference vaccine or street strains. And the four strains were genotype 1 according to the multi-sequence and phylogenetic analyses, which possessed the distinct district characteristics of China. Therefore, these four rabies viruses are likely to be street viruses already existing in the natural world.
Animals
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China
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Deer
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Disease Reservoirs
;
virology
;
Dogs
;
virology
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Ferrets
;
virology
;
Genome, Viral
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Humans
;
Mice
;
Molecular Sequence Data
;
Phylogeny
;
Rabies
;
virology
;
Rabies virus
;
chemistry
;
classification
;
genetics
;
isolation & purification
;
Sequence Analysis, DNA
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Sequence Homology, Amino Acid
;
Viral Proteins
;
chemistry
;
genetics
9.Establishment of allele-specific diagnostic PCR method for identification of antlers.
Xueyong WANG ; Chunsheng LIU ; Rong ZHANG ; Luqi HUANG ; Guanghong CUI
China Journal of Chinese Materia Medica 2009;34(23):3013-3016
OBJECTIVETo establish a convenient, quick and accurate molecular method for the identification of crude drugs of antlers due to the difficult discrimination between the genuine antler and its adulterants.
METHODAccording to the alignment analysis of full length sequences of Cyth gene from closely relate species of Cervus, one pair of allele-specific diagnostic PCR primers was designed. Factors such as annealing temperature, dosage of polymerase, times of cycles and dosage of template DNA that influence the PCR results were also investigated.
RESULTBased on the study mentioned above, about 323 bp positive band was amplified under the annealing temperature of 65 degrees C in the total volume of 25 microL PCR reaction using the genuine antler DNA as the template. Sequencing results proved that the positive band was the fragment of Cytb gene from both C. elaphus Linnaeus and C. nippon Temminck.
CONCLUSIONThe established method, with higher specificity and reproducibility, could accurately differentiate genuine antler from its adulterants and would be widely used in Cervus related Chinese crude drugs' identification.
Alleles ; Animals ; Antlers ; chemistry ; China ; DNA Primers ; genetics ; Deer ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Species Specificity
10.Genetic variability of the prion protein gene (PRNP) in wild ruminants from Italy and Scotland.
Simone PELETTO ; Matteo PERUCCHINI ; Cristina ACIN ; Mark P DALGLEISH ; Hugh W REID ; Roberto RASERO ; Paola SACCHI ; Paula STEWART ; Maria CARAMELLI ; Ezio FERROGLIO ; Elena BOZZETTA ; Daniela MELONI ; Riccardo ORUSA ; Serena ROBETTO ; Silvia GENNERO ; Wilfred GOLDMANN ; Pier Luigi ACUTIS
Journal of Veterinary Science 2009;10(2):115-120
The genetics of the prion protein gene (PRNP) play a crucial role in determining the relative susceptibility to transmissible spongiform encephalopathies (TSEs) in several mammalian species. To determine the PRNP gene variability in European red deer (Cervus elaphus), roe deer (Capreolus capreolus) and chamois (Rupicapra rupicapra), the PRNP open reading frame from 715 samples was analysed to reveal a total of ten single nucleotide polymorphisms (SNPs). In red deer, SNPs were found in codons 15, 21, 59, 78, 79, 98, 136, 168 and 226. These polymorphisms give rise to 12 haplotypes, and one of which is identical to the PRNP of American wapiti (Rocky Mountain elk, Cervus elaphus nelsoni). One silent mutation at codon 119 was detected in chamois and no SNPs were found in roe deer. This analysis confirmed that European wild ruminants have a PRNP genetic background that is compatible with TSE susceptibility, including chronic wasting disease.
Animals
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Base Sequence
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DNA/chemistry/genetics
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Deer/*genetics
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Genetic Predisposition to Disease
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Genetic Variation
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Haplotypes
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Italy
;
Molecular Sequence Data
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Phylogeny
;
Polymerase Chain Reaction/veterinary
;
Polymorphism, Single Nucleotide
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Prion Diseases/*genetics
;
Prions/*genetics
;
Scotland
;
Sequence Analysis, DNA

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