1.Validation of Reference Genes for Quantitative Real-Time PCR in Bovine PBMCs Transformed and Non-transformed by Theileria annulata.
Hongxi ZHAO ; Junlong LIU ; Youquan LI ; Congshan YANG ; Shuaiyang ZHAO ; Juan LIU ; Aihong LIU ; Guangyuan LIU ; Hong YIN ; Guiquan GUAN ; Jianxun LUO
The Korean Journal of Parasitology 2016;54(1):39-46
Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (β-actin), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.
Animals
;
B-Lymphocytes/parasitology
;
Cattle
;
Cell Line
;
Cells/*parasitology
;
Cells, Cultured
;
Gene Expression Profiling
;
Host-Parasite Interactions/*genetics
;
Real-Time Polymerase Chain Reaction/*veterinary
;
Reproducibility of Results
;
Signal Transduction/*genetics
;
Theileria annulata/physiology
;
Theileriasis/*physiopathology
2.Prevalence and Associated Risk Factors of Toxocara vitulorum Infections in Buffalo and Cattle Calves in Three Provinces of Central Cambodia.
Pierre DORNY ; Brecht DEVLEESSCHAUWER ; Valerie STOLIAROFF ; Meas SOTHY ; Rortana CHEA ; Bunthon CHEA ; Hor SOURLOING ; Sum SAMUTH ; Seth KONG ; Koemseang NGUONG ; San SORN ; Davun HOLL ; Jozef VERCRUYSSE
The Korean Journal of Parasitology 2015;53(2):197-200
The prevalence and associated risk factors of Toxocara vitulorum infection in buffalo and cattle calves was studied in 3 provinces in central Cambodia. Fecal samples were collected from 517 calves between the age of 1-15 weeks and processed for nematode egg counts by a modified McMaster method. A total of 64 calves were found to excrete T. vitulorum eggs in their feces (12.4%; 95% exact CI: 9.7-15.5). The mean fecal egg count was 2,798 EPG (SD=16,351; range=0-224,400). A multivariable generalized linear mixed model showed higher odds of T. vitulorum infection for buffalo versus cattle, for animals aged 4-8 weeks versus younger and older ones, and for animals with strongyle infection. There was no association with fecal consistency. Farmers should be aware of the potential impact of T. vitulorum, and treat their calves at the age of 2-3 weeks with anthelmintics such as benzimidazoles or pyrantel.
Animals
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Buffaloes
;
Cambodia/epidemiology
;
Cattle
;
Cattle Diseases/*epidemiology/*parasitology/physiopathology
;
Prevalence
;
Toxocara/isolation & purification/physiology
;
Toxocariasis/epidemiology/*parasitology/physiopathology
3.Ixodid Tick Infestation in Cattle and Wild Animals in Maswa and Iringa, Tanzania.
You Shine KWAK ; Tae Yun KIM ; Sung Hyun NAM ; In Yong LEE ; Hyung Pyo KIM ; Simon MDUMA ; Julius KEYYU ; Robert FYUMAGWA ; Tai Soon YONG
The Korean Journal of Parasitology 2014;52(5):565-568
Ticks and tick-borne diseases are important in human and livestock health worldwide. In November 2012, ixodid ticks were collected and identified morphologically from cattle and wild animals in the Maswa district and Iringa urban, Tanzania. Amblyomma gemma, A. lepidum, and A. variegatum were identified from Maswa cattle, and A. variegatum was the predominant species. A. marmoreum, Hyalomma impeltatum, and Rhipicephalus pulchellus were identified from Iringa cattle in addition to the above 3 Amblyomma species, and A. gemma was the most abundant species. Total 4 Amblyomma and 6 Rhipicephalus species were identified from wild animals of the 2 areas. A. lepidum was predominant in Maswa buffaloes, whereas A. gemma was predominant in Iringa buffaloes. Overall, A. variegatum in cattle was predominant in the Maswa district and A. gemma was predominant in Iringa, Tanzania.
Animals
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Animals, Wild
;
Cattle
;
Cattle Diseases/epidemiology/*parasitology
;
*Ixodidae
;
Tanzania/epidemiology
;
Tick Infestations/epidemiology/*veterinary
4.Recent Situation of Taeniasis in Mongolia (2002-2012).
Anu DAVAASUREN ; Temuulen DORJSUREN ; Tetsuya YANAGIDA ; Yasuhito SAKO ; Kazuhiro NAKAYA ; Abmed DAVAAJAV ; Gurbadam AGVAANDARAM ; Tsatsral ENKHBAT ; Battsetseg GONCHIGOO ; Nyamkhuu DULMAA ; Gantigmaa CHULUUNBAATAR ; Akira ITO
The Korean Journal of Parasitology 2014;52(2):211-214
Epidemiological situation of taeniasis in Mongolia was assessed based on mitochondrial DNA identification of the parasite species. Multiplex PCR was used on a total of 194 proglottid specimens of Taenia species and copro-PCR and loop-mediated isothermal amplification (LAMP) assays were utilized for detection of copro-DNA of 37 fecal samples from taeniasis patients submitted to the Mongolian National Center for Communicable Diseases (NCCD) from 2002 to 2012. In addition, 4 out of 44 calcified cysts in beef kept in formalin since 2003 were evaluated for histopathological confirmation of cattle cysticercosis. All proglottid specimens and stool samples were confirmed to be Taenia saginata by multiplex PCR and by copro-PCR and LAMP, respectively. Cysts collected from cattle were morphologically confirmed to be metacestodes of Taenia species. T. saginata taeniasis was identified from almost all ages from a 2-year-old boy up to a 88-year-old woman and most prominently in 15-29 age group (37%, 74/198) followed by 30-44 age group (34.8%, 69/198 ) from 15 of Mongolia's 21 provinces, while cattle cysticerci were found from 12 provinces. The highest proportion of taeniasis patients was in Ulaanbaatar, the capital of Mongolia.
Adolescent
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Adult
;
Aged
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Aged, 80 and over
;
Animals
;
Cattle/parasitology
;
Child
;
Child, Preschool
;
Cysticercosis/*epidemiology/parasitology
;
DNA, Helminth/*genetics
;
DNA, Mitochondrial/*genetics
;
Feces/parasitology
;
Female
;
Geography
;
Humans
;
Male
;
Meat/parasitology
;
Middle Aged
;
Mitochondria/genetics
;
Mongolia/epidemiology
;
Neglected Diseases/epidemiology
;
Nucleic Acid Amplification Techniques/veterinary
;
Questionnaires
;
Taenia saginata/*genetics
;
Taenia solium/genetics
;
Taeniasis/*epidemiology/parasitology
;
Young Adult
5.Sequence Analysis of cytb Gene in Echinococcus granulosus from Western China.
Xiuqin ZHONG ; Ning WANG ; Dandan HU ; Jiahai WANG ; Tianyu LIU ; Xiaobin GU ; Shuxian WANG ; Xuerong PENG ; Guangyou YANG
The Korean Journal of Parasitology 2014;52(2):205-209
Echinococcus granulosus is the causative agent of cystic echinococcosis with medical and veterinary importance in China. Our main objective was to discuss the genotypes and genetic diversity of E. granulosus present in domestic animals and humans in western China. A total of 45 hydatid cyst samples were collected from sheep, humans, and a yak and subjected to an analysis of the sequences of mitochondrial cytochrome b (cytb) gene. The amplified PCR product for all samples was a 1,068 bp band. The phylogenetic analysis showed that all 45 samples were identified as E. granulosus (genotype G1). Ten haplotypes were detected among the samples, with the main haplotype being H1. The haplotype diversity was 0.626, while the nucleotide diversity was 0.001. These results suggested that genetic diversity was low among our samples collected from the west of China based on cytb gene analysis. These findings may provide more information on molecular characteristics of E. granulosus from this Chinese region.
Animals
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Animals, Domestic/parasitology
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Base Composition
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Base Sequence
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Cattle/*parasitology
;
China
;
Cytochromes b/*genetics
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DNA, Helminth/genetics
;
Echinococcosis
;
Echinococcus granulosus/classification/*genetics
;
Genetic Variation
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Haplotypes/genetics
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Humans
;
Mitochondria/genetics
;
Molecular Sequence Data
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Phylogeny
;
Polymerase Chain Reaction/*veterinary
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Sequence Alignment
;
Sequence Analysis, DNA
;
Sheep/*parasitology
;
Tibet
6.Genetic Diversity of Echinococcus granulosus in Center of Iran.
Nader PESTECHIAN ; Ahmad HOSSEINI SAFA ; Mohammadhasan TAJEDINI ; Mohammad ROSTAMI-NEJAD ; Mohammad MOUSAVI ; Hosseinali YOUSOFI ; Shaghayegh HAGHJOOY JAVANMARD
The Korean Journal of Parasitology 2014;52(4):413-418
Hydatid cyst caused by Echinococcus granulosus is one of the most important parasitic diseases around the world and many countries in Asia, including Iran, are involved with this infection. This disease can cause high mortality in humans as well as economic losses in livestock. To date, several molecular methods have been used to determine the genetic diversity of E. granulosus. So far, identification of E. granulosus using real-time PCR fluorescence-based quantitative assays has not been studied worldwide, also in Iran. Therefore, the aim of this study was to investigate the genetic diversity of E. granulosus from center of Iran using real-time PCR method. A total of 71 hydatid cysts were collected from infected sheep, goat, and cattle slaughtered in Isfahan, Iran during 2013. DNA was extracted from protoscolices and/or germinal layers from each individual cyst and used as template to amplify the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) (420 bp). Five cattle isolates out of 71 isolates were sterile and excluded from further investigation. Overall, of 66 isolates, partial sequences of the cox1 gene of E. granulosus indicated the presence of genotypes G1 in 49 isolates (74.2%), G3 in 15 isolates (22.7%), and G6 in 2 isolates (3.0%) in infected intermediate hosts. Sixteen sequences of G1 genotype had microgenetic variants, and they were compared to the original sequence of cox1. However, isolates identified as G3 and G6 genotypes were completely consistent with original sequences. G1 genotype in livestock was the dominant genotype in Isfahan region, Iran.
Animals
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Cattle
;
Cluster Analysis
;
DNA, Helminth/chemistry/genetics
;
Echinococcosis/parasitology/*veterinary
;
Echinococcus granulosus/*classification/*genetics/isolation & purification
;
Electron Transport Complex IV/genetics
;
*Genetic Variation
;
Genotype
;
Goats
;
Iran
;
Phylogeny
;
Real-Time Polymerase Chain Reaction
;
Sequence Analysis, DNA
;
Sheep
7.Current Status of Taeniasis and Cysticercosis in Vietnam.
Nguyen VAN DE ; Thanh Hoa LE ; Phan Thi Huong LIEN ; Keeseon S EOM
The Korean Journal of Parasitology 2014;52(2):125-129
Several reports on taeniasis and cysticercosis in Vietnam show that they are distributed in over 50 of 63 provinces. In some endemic areas, the prevalence of taeniasis was 0.2-12.0% and that of cysticercosis was 1.0-7.2%. The major symptoms of taeniasis included fidgeted anus, proglottids moving out of the anus, and proglottids in the feces. Clinical manifestations of cysticercosis in humans included subcutaneous nodules, epileptic seizures, severe headach, impaired vision, and memory loss. The species identification of Taenia in Vietnam included Taenia asiatica, Taenia saginata, and Taenia solium based on combined morphology and molecular methods. Only T. solium caused cysticercosis in humans. Praziquantel was chosen for treatment of taeniasis and albendazole for treatment of cysticercosis. The infection rate of cysticercus cellulosae in pigs was 0.04% at Hanoi slaughterhouses, 0.03-0.31% at provincial slaughterhouses in the north, and 0.9% in provincial slaughterhouses in the southern region of Vietnam. The infection rate of cysticercus bovis in cattle was 0.03-2.17% at Hanoi slaughterhouses. Risk factors investigated with regard to transmission of Taenia suggested that consumption of raw meat (eating raw meat 4.5-74.3%), inadequate or absent meat inspection and control, poor sanitation in some endemic areas, and use of untreated human waste as a fertilizer for crops may play important roles in Vietnam, although this remains to be validated.
Albendazole/*therapeutic use
;
Animals
;
Anthelmintics/therapeutic use
;
Antiprotozoal Agents/therapeutic use
;
Cattle/parasitology
;
Cattle Diseases/parasitology
;
Cysticercosis/*drug therapy/*epidemiology/parasitology/transmission
;
Feces/parasitology
;
Humans
;
Meat/parasitology
;
Praziquantel/*therapeutic use
;
Raw Foods/adverse effects
;
Risk Factors
;
Swine/parasitology
;
Swine Diseases/parasitology
;
Vietnam/epidemiology
8.Development of a Disperse Dye Immunoassay Technique for Detection of Antibodies against Neospora caninum in Cattle.
Fatemeh SELAHI ; Mehdi NAMAVARI ; Mohammad Hossein HOSSEINI ; Maryam MANSOURIAN ; Yahya TAHAMTAN
The Korean Journal of Parasitology 2013;51(1):129-132
In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.
Animals
;
Antibodies, Protozoan/*blood
;
Cattle
;
Cattle Diseases/*diagnosis/parasitology
;
Coccidiosis/diagnosis/parasitology/*veterinary
;
Diagnostic Tests, Routine/*methods
;
Female
;
Immunoassay/methods
;
Neospora/*immunology
;
Staining and Labeling/methods
;
Veterinary Medicine/*methods
9.Morphology and histology of the adult Paramphistomum gracile Fischoeder, 1901.
Busaba PANYARACHUN ; Arin NGAMNIYOM ; Prasert SOBHON ; Panat ANURACPREEDA
Journal of Veterinary Science 2013;14(4):425-432
In the present study, we evaluated the histological morphology of the adult Paramphistomum (P.) gracile. Adult flukes with bodies 5~15 mm in length and 2~7 mm in width were subjected to histological analysis. Longitudinal and transversal serial-sections were stained with hematoxylin and eosin, and examined. The body surface and longitudinal section of P. gracile were also assessed using scanning electron microscopy. In this species, the anterior sucker and posterior sucker (acetabulum) were present on an anterior and posterior part of the body, respectively. The major folds were located in the areas of the anterior sucker, genital canal, and posterior sucker. The fluke membrane was spineless at the tegument surface and in the tegument tissue. Histological data showed structural-systematic characteristics of the digestive tract, reproductive tract, excretory tract, copulatory organs, connective tissues, and muscle tissues. We attempted to elucidate the histological characteristics of P. gracile that might increase the knowledge and understanding of rumen fluke morphology.
Animals
;
Cattle
;
Cattle Diseases/*parasitology/pathology
;
Female
;
Male
;
Microscopy, Electron, Scanning/veterinary
;
Rumen/parasitology
;
Stomach Diseases/parasitology/pathology/*veterinary
;
Thailand
;
Trematoda/*anatomy & histology/isolation & purification/ultrastructure
;
Trematode Infections/parasitology/pathology/*veterinary
10.Study of infection of Echinococcus granulosus in yak in spring and its potential role in transmission of cystic Echinococcosis in Rangtang County of Sichuan, China.
Huan Huan HU ; Wei Ping WU ; Li Ying WANG ; Qian WANG ; Yan HUANG ; Ya Yi GUAN
Biomedical and Environmental Sciences 2013;26(3):226-229
Animals
;
Cattle
;
China
;
epidemiology
;
Echinococcosis
;
epidemiology
;
parasitology
;
transmission
;
Echinococcus granulosus
;
pathogenicity
;
Humans
;
Seasons

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