1.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
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Male
2.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
3.Research on the identification of penis et testis cervi with molecular taxonomy.
Shuang-yan TANG ; Wen FU ; Yong-jiu CHEN ; Jian-yun WANG ; Xu JIANG ; Ya-ping ZHANG
China Journal of Chinese Materia Medica 2002;27(8):573-575
OBJECTIVETo make the kit with witch to identify Penis et Testis Cervi with molecular taxonomy.
METHODThe mtDNA of sika and red deer from different areas was amplified by PCR and sequenced. Compared with the mtDNA of bovine and horse from witch the false medicines were made, characteristic segments of deer were found. We selected one as the species distinctive PCR primer of deer.
RESULTThe kit made up with this primer and related reagents could be used to discern Penis et Testis Cervi from the false medicine.
CONCLUSIONIt is a scientific, steady, accurate and convenient way to identify Penis et Testis Cervi with molecular taxonomy.
Animals ; Cattle ; genetics ; DNA ; genetics ; DNA Primers ; DNA, Mitochondrial ; genetics ; Deer ; classification ; genetics ; Drug Contamination ; Horses ; genetics ; Male ; Materia Medica ; chemistry ; Penis ; chemistry ; Testis ; chemistry
4.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
5.Identification of single-nucleotide polymorphisms of the prion protein gene in sika deer (Cervus nippon laiouanus).
Hyun Jeong JEONG ; Joong Bok LEE ; Seung Yong PARK ; Chang Seon SONG ; Bo Sook KIM ; Jung Rae RHO ; Mi Hyun YOO ; yung Hoon JEONG ; Yong Sun KIM ; In Soo CHOI
Journal of Veterinary Science 2007;8(3):299-301
Polymorphisms of the prion protein gene (PRNP) havebeen detected in several cervid species. In order toconfirm the genetic variations, this study examined theDNA sequences of the PRNP obtained from 33 captivesika deer (Cervus nippon laiouanus) in Korea. A total ofthree single-nucleotide polymorphisms (SNPs) at codons100, 136 and 226 in the PRNP of the sika deer wereidentified. The polymorphic site located at codon 100 hasnot been reported. The SNPs detected at codons 100 and226 induced amino acid substitutions. The SNP at codon136 was a silent mutation that does not induce any aminoacid change. The genotype and allele frequencies weredetermined for each of the SNPs.
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 Variation
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Molecular Sequence Data
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Polymerase Chain Reaction/veterinary
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Polymorphism, Single Nucleotide
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Prions/*genetics
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Sequence Analysis, DNA
6.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
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genetics
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DNA Primers
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genetics
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Deer
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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
7.Mycobacterium bovis Infection in a Farmed Elk in Korea.
Jae Hoon KIM ; Hyun Joo SOHN ; Kyung Il KANG ; Won Il KIM ; Jong Sam AN ; Young Hwa JEAN
Journal of Veterinary Science 2002;3(3):163-166
A case of tuberculosis is reported in an eight-year-old, male, elk (Cervus elaphus nelsoni). The elk showed severe coughing, respiratory distress, abdominal breathing, anorexia, and severe progressive emaciation in the elk farm. At necropsy, the elk appeared in poor body condition. Mild enlargement of retropharyngeal and submandibular lymph node was observed in the head. Diffuse fibrinous pleuritis and purple red lobar pneumonia were found in the thorax. Well demarcated numerous dark yellow discrete or confluent nodules from 0.3 to 2 cm in diameter were scattered in the whole lung. Bronchial and mediastinal lymph nodes were also enlarged. Histopathologically, lungs had typical classical tuberculous granulomas, multiple abscesses, and numerous macrophages and Langhans giant cells infiltration in alveolar lumen. In the lymph nodes, there were small clusters of necrosis and infiltration of numerous macrophages, epithelioid cells, and Langhans giant cells. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. According to this study, there are differences of the histopathologic lesions and the numbers of acid-fast bacilli in the lesions between this elk and cattle. Mycobacterium bovis was confirmed as a causative agent in this elk using bacterial isolation, biochemical characteristics, and PCR technique. The isolate was negative for niacin test, nitrate reductase, and pyrazinamidase. This is a first report for bovine tuberculosis of farmed elk in Asia.
Animals
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DNA, Bacterial/chemistry/genetics
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Deer/*microbiology
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Fatal Outcome
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Korea
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Lung/microbiology/pathology
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Male
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Mycobacterium bovis/genetics/*isolation&purification
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Polymerase Chain Reaction
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Tuberculosis/microbiology/*veterinary
8.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
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Molecular Sequence Data
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Phylogeny
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Polymerase Chain Reaction/veterinary
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Polymorphism, Single Nucleotide
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Prion Diseases/*genetics
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Prions/*genetics
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Scotland
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Sequence Analysis, DNA
9.Molecular fingerprinting of clinical isolates of Mycobacterium bovis and Mycobacterium tuberculosis from India by restriction fragment length polymorphism (RFLP).
Sandeep Kumar SINGH ; Rishendra VERMA ; Devendra H SHAH
Journal of Veterinary Science 2004;5(4):331-335
Forty mycobacterial strains comprising clinical Indian isolates of Mycobacterium tuberculosis (28 field isolates +1H37 Rv) and Mycobacterium bovis (10 field isolates +1 AN5) were subjected to restriction fragment length polymorphism analysis (RFLP) using IS6110 and IS1081 probes. Most of these strains originated from dairy cattle herd and human patients from Indian Veterinary research Institute (IVRI) campus isolated from the period of 1986 to 2000. Our study showed presence of 8 copies of IS6110 in most of the M.tuberculosis (96.6%) strains irrespective of their origin with the exception of one M.tuberculosis strain with presence of an extra copy (3.4%). All M.bovis strains showed a single copy of IS6110 on the characteristic 1.9kb restriction fragment. RFLP analysis with IS1081 invariably showed the presence of 5 copies in all isolates of M.bovis and M.tuberculosis at the same chromosomal location. Similarity of IS6110 RFLP fingerprints of M.tuberculosis strains from animals and human suggested the possibility of dissemination of single M.tuberculosis strain among animals as well as human. It was not possible to discriminate within the isolates of either M.tuberculosis or M.bovis, when IS1081 was used as target sequence. The IS6110 RFLP is a valuable tool for disclosing transmission chain of M. tuberculosis and M. bovis among humans as well as animals
Animals
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Bacterial Typing Techniques
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Cattle
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DNA Fingerprinting/*veterinary
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DNA, Bacterial/*genetics
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Deer
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Humans
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India/epidemiology
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Mycobacterium bovis/classification/*genetics/isolation&purification
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Mycobacterium tuberculosis/classification/*genetics/isolation & purification
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Polymerase Chain Reaction/veterinary
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Polymorphism, Restriction Fragment Length
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Zoonoses/epidemiology
10.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
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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
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United States