1.Re-transmissibility of mouse-adapted ME7 scrapie strain to ovine PrP transgenic mice
Joshua Adekunle BABALOLA ; Jong Mu KIM ; Yun Jung LEE ; Jeong Ho PARK ; Hong Seok CHOI ; Yeong Gon CHOI ; Eun Kyoung CHOI ; Yong Sun KIM
Journal of Veterinary Science 2019;20(2):e8-
Scrapie is a mammalian transmissible spongiform encephalopathy or prion disease that predominantly affects sheep and goats. Scrapie has been shown to overcome the species barrier via experimental infection of other rodents. To confirm the re-transmissibility of the mouse-adapted ME7 scrapie strain to ovine prion protein (PrP) transgenic mice, mice of an ovinized transgenic mouse line carrying the Suffolk sheep PrP gene that contained the A₁₃₆ R₁₅₄ Q₁₇₁/ARQ allele were intracerebrally inoculated with brain homogenates obtained from terminally ill ME7-infected C57BL/6J mice. Herein, we report that the mouse-adapted ME7 scrapie strain was successfully re-transmitted to the transgenic mice expressing ovine PrP. In addition, we observed changes in the incubation period, glycoform profile, and pattern of scrapie PrP (PrP(Sc)) deposition in the affected brains. PrP(Sc) deposition in the hippocampal region of the brain of 2nd-passaged ovine PrP transgenic mice was accompanied by plaque formation. These results reveal that the mouse-adapted ME7 scrapie strain has the capacity to act as a template for the conversion of ovine normal monomeric precursors into a pathogenic form in ovine PrP transgenic mice. The change in glycoform pattern and the deposition of plaques in the hippocampal region of the brain of the 2nd-passaged PrP transgenic mice are most likely cellular PrP species dependent rather than being ME7 scrapie strain encoded.
Alleles
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Animals
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Brain
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Gliosis
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Goats
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Humans
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Mice
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Mice, Transgenic
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Plaque, Amyloid
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Prion Diseases
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PrPSc Proteins
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Rodentia
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Scrapie
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Sheep
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Terminally Ill
2.A specific PCR assay and a nested PCR assay for the screening of ovine pulmonary adenomatosis.
Jun-Rong LUO ; Xiao-Hui LIU ; Qun-Ying YU ; Shu-Qing ZHANG ; Jian-Hua ZHOU ; Xue-En MA
Chinese Journal of Virology 2009;25(2):125-130
Ovine pulmonary adenomatosis (OPA) is a naturally occurring contagious lung tumor of sheep which was caused by an exogenous retrovirus of sheep, jaagsiekte retrovirus (JSRV). Although no specific circulating antibodies against the virus coud be detected in infected sheep, exogenous JSRV proviral DNA sequences (exJSRV) and JSRV RNA transcripts could be detected in lung tumors, lymphoreticular system and peripheral blood mononuclear cells (PBMC) from sheep affected by OPA. The sheep genome carried 15 to 20 copies of endogenous retrovirus loci (enJSRV) that were similar to JSRV in structural genes but the divergene in U3. Therefore, primers specific for the U3 sequences of exJSRV were designed for the specific PCR and nested PCR (n-PCR). Sensitivity between specific PCR assay and n-PCR assay was compared by using serial dilutions of positive plasmid pJSRV-LTR in a background of 700ng sheep genome DNA. Sensitivity of n-PCR was ten-fold higher than specific PCR. The n-PCR was only available in blood test for detection of JSRV infected sheep and might be useful in epidemiological studies and disease control of OPA.
Animals
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Base Sequence
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Clinical Laboratory Techniques
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DNA, Viral
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analysis
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Endogenous Retroviruses
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genetics
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Jaagsiekte sheep retrovirus
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genetics
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Lung Neoplasms
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virology
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Molecular Sequence Data
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Polymerase Chain Reaction
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Pulmonary Adenomatosis, Ovine
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virology
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Reverse Transcriptase Polymerase Chain Reaction
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Sensitivity and Specificity
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Sequence Analysis, DNA
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Sheep
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Sheep Diseases
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virology
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Virus Cultivation
3.Infection Status of Hydatid Cysts in Humans and Sheep in Uzbekistan.
Sung Tae HONG ; Yan JIN ; Khikmat ANVAROV ; Abdukhakim KHADJIBAEV ; Samin HONG ; Yusufjon AHMEDOV ; Utkir OTABOEV
The Korean Journal of Parasitology 2013;51(3):383-385
Uzbekistan is endemic of cystic echinococcosis (CE). In order to estimate endemicity of CE, we collected data from emergency surgery due to CE in 2002-2010 and also investigated the prevalence of hydatid cysts in the liver and lungs of sheep at an abattoir in Uzbekistan from July 2009 to June 2010. In 14 emergency hospitals, 8,014 patients received surgical removal or drainage of CE during 2002-2010, and 2,966 patients were found in 2010. A total of 22,959 sheep were grossly examined of their liver and lungs, and 479 (2.1%) and 340 (1.5%) of them were positive for the cyst in the liver and lungs, respectively. Echinococcus granulosus is actively transmitted both to humans and sheep, and CE is a zoonotic disease of public health priority in Uzbekistan.
Animals
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Echinococcosis/epidemiology/*parasitology
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Humans
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Sheep
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Sheep Diseases/epidemiology/*parasitology
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Uzbekistan/epidemiology
4.Diprosopus, craniorachischisis, arthrogryposis, and other associated anomalies in a stillborn lamb.
Cihan KACR ; Kadir OZCAN ; Ismet TAKCI ; Kutlay GURBULAK ; Hasan OZEN ; Musa KARAMAN
Journal of Veterinary Science 2008;9(4):429-431
Congenital malformations with multiple anomalies have been described infrequently in the veterinary literature. A stillborn male crossbred lamb with diprosopus, craniorachischisis, and arthrogryposis was examined macroscopically and histopathologically in this study. The left head was smaller than the right head. Micrencephaly, agnathia, and a rudimentary tongue, which was adherent to the palate, were present in the left head. Micrencephaly, brachygnathia superior, and cleft palate were present in the right head. Cerebellar agenesis and spinal cord hypoplasia were observed. The cerebrums and the spinal cord were covered with a tapering membranous structure. Neural and dermal tissues were noted to intervene upon microscopic examination of this structure. Disorganization of neurons was observed in both cerebrums, though it was more severe in the left one. This case demonstrates many congenital defects occurring together in a lamb.
Animals
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Congenital Abnormalities/pathology/*veterinary
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Male
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Sheep
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Sheep Diseases/*congenital/pathology
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Stillbirth
5.Expression and refolding of OLA Ⅰ protein with peptides derived from sheeppox virus.
Zhanhong WANG ; Zhixun ZHAO ; Guohua WU ; Yang DENG ; Guoqiang ZHU ; Fangyan ZHAO ; Zengjun LU ; Qiang ZHANG
Chinese Journal of Biotechnology 2022;38(1):139-147
The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β2m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a SuperdexTM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
Animals
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Capripoxvirus
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Epitopes, T-Lymphocyte/genetics*
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Peptides/genetics*
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Poxviridae Infections
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Sheep
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Sheep Diseases
6.Surgical treatment for different forms of hernias in sheep and goats.
Fahd A AL-SOBAYIL ; Ahmed F AHMED
Journal of Veterinary Science 2007;8(2):185-191
Sheep and goats are frequently presented with different forms of hernias to veterinary clinics. The aim of this study is to investigate the outcome of the surgical treatment of abdominal, umbilical, inguinal and scrotal hernias in sheep and goats. Fifty-eight clinical cases (sheep = 44, goat = 14) were presented to the Veterinary Teaching Hospital, College of Agriculture and Veterinary Medicine, Qassim University, Saudi Arabia from September, 2003 to September, 2006. These animals had abdominal (sheep = 30, goat = 10), umbilical (sheep = 6, goat = 4), inguinal (sheep = 7) and scrotal (sheep = 1) hernias. All the cases of hernias in sheep and goats were subjected to full study including the history of the case, classification of hernias, the size of the hernial ring, surgical repair of the hernias, adhesions between the hernial sacs in each case, the postoperative care and follow up of the cases. The results revealed that gender had an effect on the incidence of hernia. The incidence of abdominal hernias was higher in females and the incidence of inguinal hernia was higher in males. There was a positive correlation between the history of hernia and the degree of adhesion. For the sheep, 26 out of 30 cases of abdominal hernia had good outcomes and the healing was excellent. There were postoperative complications in 4 ewes. For the goats, there were slight swellings at the site of operation in 2 out of 10 cases of abdominal hernia, while the remaining 8 cases had good outcomes. There was one case of umbilical hernia with an umbilical abscess that had broken down with sepsis formation at the surgical site. In conclusion, the success rates of surgical treatment for all types of hernias were very high and there were no significant differences in the success rates among the different types of hernias in both sheep and goats. The types of suture materials and the types of hernias had no significant effect on the outcome of the surgical treatment.
Animals
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Female
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Goat Diseases/pathology/*surgery
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Goats
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Hernia/pathology/surgery/*veterinary
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Male
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Sheep
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Sheep Diseases/pathology/*surgery
7.Comparative efficacy of standard AGID, CCIE and competitive ELISA for detecting bluetongue virus antibodies in indigenous breeds of sheep and goats in Rajasthan, India.
Journal of Veterinary Science 2005;6(1):77-79
The sero-prevalence of antibodies against blue tongue virus (BTV) in 408 local breeds of sheep in Rajasthan state in India was investigated using standard agar gel immunodiffusion (AGID) test. Maximum seropositivities of 11.3% (13/115), 10.7% (13/121), 7.1% (11/155) and 5.9% (1/17) were recorded in the Chokla, Magra, Nali and Pugal breeds, respectively. Out of 107 goat serum samples, 6 (5.6%) were AGID positive. The performance of the standard AGID, counter current immuno-electrophoresis (CCIE) and the competitive enzyme-linked immunosorbent assay (cELISA) for the detection of serum antibody against BTV in indigenous breeds of sheep were compared. Out of 178 sheep serum samples tested, 17 (9.5%), 22 (12.3%) and 54 (30.3%) were positive for group-specific bluetongue antibodies by AGID, CCIE and cELISA, respectively. There was appreciable difference in the seroprevalence detected by AGID, CCIE and cELISA in clinically healthy and diseased sheep with regard to relative sensitivities and specificities of the tests with cELISA being highly sensitive and specific followed by CCIE and AGID test. It was concluded that these indigenous breeds of sheep may be a potential reservoir of BTV infection and cELISA should be routinely used for the detection of antibodies against BTV in these local breeds of sheep.
Animals
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Antibodies, Viral/*analysis
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Bluetongue/*diagnosis/*epidemiology
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Counterimmunoelectrophoresis
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Enzyme-Linked Immunosorbent Assay
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Goat Diseases/*diagnosis
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Goats
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Immunodiffusion
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India/epidemiology
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Prevalence
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Sheep
8.Morphological Characteristics of Normal and Gynandromorphic Hyalomma asiaticum Schulze and Schlottke, 1930.
Ze CHEN ; You Quan LI ; Qiao Yun REN ; Jin LUO ; Yonghong HU ; Kai LI ; Guang Yuan LIU ; Jian Xun LUO ; Jingze LIU ; Hong YIN
The Korean Journal of Parasitology 2015;53(3):361-364
Gynandromorphic ticks are extremely rare, and often attract parasitologists' attention. During our examination of tick specimens, an engorged gynandromorph of Hyalomma asiaticum was noticed. This is the first record of gynandromorphic ticks from China. In this study, several important morphological structures of normal and gynandromorphic H. asiaticum were analyzed. Comparing to the normal H. asiaticum, the gynandromorphic specimen was a typical bipartite protogynander. Its right side showed normal female characteristics, whereas the left side had normal male traits. Different from other gynandromorphic ticks containing 1 anus, this tick reported here had 2 complete anuses, and the anus of the male part had a single adanal plate.
Animals
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Chimera/*anatomy & histology/genetics
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China
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Female
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Ixodidae/*anatomy & histology/genetics
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Male
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Sheep
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Sheep Diseases/*parasitology
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Tick Infestations/parasitology/*veterinary
9.In vitro protoscolicidal effects of hypertonic glucose on protoscolices of hydatid cyst.
Seyed Vahid HOSSEINI ; Kurosh GHANBARZADEH ; Jahra BARZIN ; Seyed Mahmoud SADJJADI ; Nader TANIDEH ; Davood MEHRABANI
The Korean Journal of Parasitology 2006;44(3):239-242
To evaluate the protoscolicidal effects of various concentrations of hypertonic glucose, live protoscolices of sheep were exposed to 10%, 15%, 25% and 50% glucose solutions. Cetrimide (0.5%), silver nitrate (0.5%) and hypertonic saline (20%) were used as positive controls, while physiological saline was used as a negative control. After 1, 2 and 5 min, the protoscolicidal effects were determined by 1% eosin. A 25% glucose solution had no significant protoscolicidal effect. However, a 50% glucose solution revealed higher protoscolicidal effect than 0.5% silver nitrate but weaker effect than 0.5% cetrimide; the effect was comparable with that of 20% hypertonic saline. The results showed that hypertonic glucose solution is highly effective in killing protoscolices of Echinococcus granulosus in vitro.
Sheep Diseases/*parasitology
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Sheep
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Glucose Solution, Hypertonic/*pharmacology
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Echinococcus granulosus/*drug effects/isolation & purification
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Echinococcosis/*parasitology
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Animals
10.Analysis of monoclonal antibody binding sites in ovine prion protein.
Yongqiang ZHANG ; Xiaodong WU ; Yonggang ZHAO ; Endong BAO ; Qinghua WANG ; Wei ZHANG ; Yutian LIU ; Zhiliang WANG
Chinese Journal of Biotechnology 2009;25(3):348-353
Binding sites of five monoclonal antibodies were obtained by reinforceable method of overlapping recombinant prion protein and synthetic peptide. Overlapping peptides of PrP core were expressed in Escherichia coli by insertion of serial PCR amplicons of ovine PrP gene fragments into pET32a. The expressed fusion peptides were then tested for the binding activity to PrP monoclonal antibodies in Western blotting. The binding sites of 5 monoclonal antibodies of ovine PrP were located respectively as follows: 2H3 in 199 aa-213 aa, 4C6, 5F11 and 7F11 in 139 aa-168 aa and 7F1 in 214 aa-227 aa. There oligo peptides were synthesized and used in ELISA test for more accurate localization of the binding sites. The binding sites of 4C6, 5F11 and 7F11 were further confirmed to be in 149 aa-158 aa. This conclusion may contribute to the research for pathogenesis and diagnostic method of scrapie and bovine transmissible spongiform encephalopathy.
Animals
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Antibodies, Monoclonal
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immunology
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metabolism
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Binding Sites, Antibody
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immunology
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Epitopes
;
immunology
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Escherichia coli
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genetics
;
metabolism
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Prion Diseases
;
diagnosis
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Prions
;
genetics
;
immunology
;
metabolism
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Recombinant Fusion Proteins
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genetics
;
immunology
;
metabolism
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Scrapie
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diagnosis
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Sheep