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3D reconstruction and capsid protein characterization of grass carp reovirus
Qin Fang,Sanket Shah,Yuyao Liang,H. Z. Zhou
Science China Life Sciences , 2005, DOI: 10.1360/062004-105
Abstract: Grass carp reovirus (GCRV) is a relatively new virus first isolated in China and is a member of the Aquareovirus genus of the Reoviridae family. Recent report of genomic sequencing showed that GCRV shared high degree of homology with mammalian reovirus (MRV). As a step of our effort to understand the structural basis of GCRV pathogenesis, we determined the three-dimensional (3D) structure of GCRV capsid at 17 resolution by electron cryomicroscopy. Each GCRV capsid has a multilayered organization, consisting of an RNAcore, an inner, middle and outer protein layer. The outer layer is made up of 200 trimers that are arranged on an incomplete T=13 icosahedral lattice. A characteristic feature of this layer is the depression resulting from the absence of trimers around the peripentonal positions, revealing the underlying trimers on the middle layer. There are 120 subunits in the inner layer arranged with T=1 symmetry. These structural features are common to other members of the Reoviridae. Moreover, SDS-PAGE analysis showed that GCRV virions contain seven structural proteins (VP1-VP7). These structural proteins have a high degree of sequence homology to MRV, consistent with the structural similarities observed in our study. The high structural similarities of isolated GCRV and MRV suggest that future structural studies focusing on GCRV entering into and replicating within its host cell are necessary in order to fully understand the structural basis of GCRV pathogenesis.
3D reconstruction and capsid protein characterization of grass carp reovirus
FANG Qin,Shah Sanket,LIANG Yuyao,Z H ZHOU,
Shah
,Sanket

中国科学C辑(英文版) , 2005,
Abstract: Grass carp reovirus (GCRV) is a relatively new virus first isolated in China and is a member of the Aquareovirus genus of the Reoviridae family. Recent report of genomic se-quencing showed that GCRV shared high degree of homology with mammalian reovirus (MRV). As a step of our effort to understand the structural basis of GCRV pathogenesis, we determined the three-dimensional (3D) structure of GCRV capsid at 17 ? resolution by electron cryomicro-scopy. Each GCRV capsid has a multilayered organization, consisting of an RNA core, an inner, middle and outer protein layer. The outer layer is made up of 200 trimers that are arranged on an incomplete T=13 icosahedral lattice. A characteristic feature of this layer is the depression re-sulting from the absence of trimers around the peripentonal positions, revealing the underlying trimers on the middle layer. There are 120 subunits in the inner layer arranged with T=1 symme-try. These structural features are common to other members of the Reoviridae. Moreover, SDS-PAGE analysis showed that GCRV virions contain seven structural proteins (VP1-VP7). These structural proteins have a high degree of sequence homology to MRV, consistent with the structural similarities observed in our study. The high structural similarities of isolated GCRV and MRV suggest that future structural studies focusing on GCRV entering into and replicating within its host cell are necessary in order to fully understand the structural basis of GCRV pathogenesis.
Protective Roles of Grass Carp Ctenopharyngodon idella Mx Isoforms against Grass Carp Reovirus  [PDF]
Limin Peng, Chunrong Yang, Jianguo Su
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0052142
Abstract: Background Myxovirus resistance (Mx) proteins are crucial effectors of the innate antiviral response against a wide range of viruses, mediated by the type I interferon (IFN-I) signaling pathway. However, the antiviral activity of Mx proteins is diverse and complicated in different species. Methodology/Principal Findings In the current study, two novel Mx genes (CiMx1 and CiMx3) were identified in grass carp (Ctenopharyngodon idella). CiMx1 and CiMx3 proteins exhibit high sequence identity (92.1%), and low identity with CiMx2 (49.2% and 49.5%, respectively) from the GenBank database. The predicted three-dimensional (3D) structures are distinct among the three isoforms. mRNA instability motifs also display significant differences in the three genes. The spatial and temporal expression profiles of three C. idella Mx genes and the IFN-I gene were investigated by real-time fluorescence quantitative RT-PCR (qRT-PCR) following infection with grass carp reovirus (GCRV) in vivo and in vitro. The results demonstrated that all the four genes were implicated in the anti-GCRV immune response, that mRNA expression of Mx genes might be independent of IFN-I, and that CIK cells are suitable for antiviral studies. By comparing expression patterns following GCRV challenge or poly(I:C) treatment, it was observed that GCRV blocks mRNA expression of the four genes. To determine the functions of Mx genes, three CiMx cDNAs were cloned into expression vectors and utilized for transfection of CIK cells. The protection conferred by each recombinant CiMx protein against GCRV infection was evaluated. Antiviral activity against GCRV was demonstrated by reduced cytopathic effect, lower virus titer and lower levels of expressed viral transcripts. The transcription of IFN-I gene was also monitored. Conclusions/Significance The results indicate all three Mx genes can suppress replication of grass carp reovirus and over-expression of Mx genes mediate feedback inhibition of the IFN-I gene.
Comparative Histomorphological Studies on Oesophagus of Catfish and Grass Carp  [PDF]
Enas A. Abd El Hafez,Doaa M. Mokhtar,Alaa Sayed Abou-Elhamd,Ahmed Hassan S. Hassan
Journal of Histology , 2013, DOI: 10.1155/2013/858674
Abstract: The present work was carried out on 40 specimens of oesophaguses of both sexes of catfish (carnivorous fish) and grass carp (herbivorous fish) in order to observe the morphological and histological differences between the two species. Oesophagus of catfish was divided into 2 parts: anterior and posterior ones. The anterior part of the oesophagus of catfish was characterized by the presence of numerous mucosal folds. It was lined by stratified epithelium with goblet cells. In addition to club cells were observed in between the stratified epithelium. Scanning electron examination of the oesophageal epithelium of catfish demonstrated the presence of microvilli and fingerprint-like microridges in the superficial cell layer. The posterior part of the oesophagus of catfish was characterized by simple columnar mucus-secreting epithelium. The oesophagus of grass carp had shown the same structure along its entire length. It consisted of less folded mucosa than that observed in the oesophagus of catfish. The epithelium was characterized by the presence of taste buds. In conclusion, the present work revealed some differences in the structure of catfish oesophagus and grass carp oesophagus. These differences are related to type of food and feeding habits of each species. 1. Introduction Nile catfish (Clarias gariepinus) is one of the most abundant and widely distributed fish in the Nile River. Catfish has a wide geographical spread, a high growth rate, resistant to handling and stress, and well appreciated in a wide number of African countries. It is considered the third important commercial fish in Egypt after tilapia and bagrids [1, 2]. It can be recognized by its long dorsal and anal fins, which gives it a rather eel-like appearance. The catfish is carnivorous in type, where tilapias are its most preferred food item especially the young ones followed by insects, crustaceans, and mollusks, respectively [3]. The grass carp or white amur (Ctenopharyngodon idella) is a large cyprinid fish. They are native in large Asian rivers such as Amur River Basin in Russia and the West River in China [4]. It is a fast growing herbivorous fish; it usually feeds on grass or other aquatic vegetations and can be grown together with other fish species. The gastrointestinal tract of grass carp consisted of oesophagus, stomach, and intestine that ends at the anus. Food enters a short, often greatly distensible esophagus leading into a thick-walled stomach [5, 6]. The histological structure of the gastrointestinal tract of numerous fish species generally consists of mucosa, submucosa,
Study of some biochemical parameters in the grass carp  [cached]
Elena Ciornea,Gabriela Dumitru,Sabina-Ioana Cojocaru
Analele ?tiin?ifice Ale Universit??ii Alexandru Ioan Cuza din Ia?i,Sectiunea II A : Genetica si Biologie Moleculara , 2009,
Abstract: The experimental investigations were devoted to some biochemical parameters (glycogen, catalase, alanine- and aspartate-aminotransferase) in three summer-old representatives of grass carp, at the level of the hepatic and muscular tissue. The results obtained evidenced significant differences between the tissues under analysis, both in the concentration of the main reserve polyglucides, as a supplier of circulating glucose, and in the enzymatic activity, in close correlation with the physiological and biochemical role of each tissue in part.
Prevalence of Copepod Ectoparasites of Grass Carp Ctenopharyngodon idella  [PDF]
Zahida Tasawar,Rabia Naseem,Masood Akhtar
Pakistan Journal of Biological Sciences , 1999,
Abstract: One hundred and twenty grass carp, Ctenopharyngodon idella, were examined over a period of one year from March 1998 to February 1999 at Government Fish Hatchery, Mian Channu, Punjab, Pakistan. Four species of copepod ectoparasites recovered were Lernaea (L.) polymorpha (Yu, 1938), L. cyprinacea (Linnaeus, 1761), L. lophiara (Harding, 1950) and L. ctenopharyngodonis. The parasitic infestation was low during the summer months when temperature ranged between 30-32 C and highest in winter months when temperature was between 13 -23 C.
THE PREVALENCE OF LERNAEID ECTOPARASITES IN GRASS CARP (CTENOPHARYNGODON IDELLA)
Z. TASAWAR, S. ZAFAR, M. H. LASHARI AND C. S. HAYAT1
Pakistan Veterinary Journal , 2009,
Abstract: The present study was conducted to investigate the prevalence of lernaeid ectoparasites in grass carp (Ctenopharyngodon idella). For this purpose, 597 fishes (Ctenopharyngodon idella) were examined for lernaeid ectoparasites at a private fish farm located in Multan, Pakistan. Four species of the genus Lernaea i.e. L. cyprinacea, L. polymorpha, L. oryzophila, and L. lophiara were recorded. It was observed that L. polymorpha had the highest (P<0.05) overall prevalence (7.54%), followed by L. cyprinacea (6.53%), Learnaea species which were not identified (2.18%), L. oryzophila and L. lophiara (0.67% each). The relationship between body weight and Lernaea infestation showed that the infection of Lernaea species was significantly (P<0.05) more prevalent in the weight group of 2501-4500g, while the parasites were not found in the weight groups of 4500-6500 and 6501-8500g. Relationship between body length and Lernaea species in fish was also calculated. According to these results, the Lernaea species had significantly (P<0.05) highest prevalence in length group of 9-14 cm and lowest in length groups of 15-20 and >20 cm.
The Primary and Subculture and Identification of Grass Carp Preadipocyte from Adipose Tissue  [PDF]
CAO Kai,LI Ying,CAI Zhi-hua
Journal of Chongqing Normal University , 2010,
Abstract: To establish the culture system of grass carp (Ctenopharyngodon idellus) preadipocyte, and redisplay the whole process of the proliferation and differentiation of grass carp preadipocytes in vitro, these experiments were carried out. The adipocytes were identified by oil red O staining, and the cells morphology was observed by inverted microscope and CCD. Grass carp selected was healthy and about 800-900 g. Primary culture of adipose tissue from grass carp abdominal cavity was done in the DMEM/F12 medium by using the tissure culture technology. All the experiments were done under the condition of 28℃,5% CO2, 20% calf serum, pH7.0-7.2.When monolayer cells occupied 70%-80% of the bottle's bottom and confluenced, the subculture was done. The results showed that the spindle cells emigrated from the tissues after 48 hours, the cell number increased and the cell morphology were mainly spindle and polygon after 3 days, cells confluenced most after 8 days and oil red O staining proved that stained cells were adipocytes. Confluenced cells could be passed at fourth passage. The cells could formed monolayer at the third day, and cytoplasm contained large quantities of lipid droplets at the eighth day. In summary, the sutiable culture system of grass carp preadipocyte was estabilished initially.
Composition, Diversity, and Origin of the Bacterial Community in Grass Carp Intestine  [PDF]
Shangong Wu, Guitang Wang, Esther R. Angert, Weiwei Wang, Wenxiang Li, Hong Zou
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0030440
Abstract: Gut microbiota has become an integral component of the host, and received increasing attention. However, for many domestic animals, information on the microbiota is insufficient and more effort should be exerted to manage the gastrointestinal bacterial community. Understanding the factors that influence the composition of microbial community in the host alimentary canal is essential to manage or improve the microbial community composition. In the present study, 16S rRNA gene sequence-based comparisons of the bacterial communities in the grass carp (Ctenopharyngodon idellus) intestinal contents and fish culture-associated environments are performed. The results show that the fish intestinal microbiota harbors many cellulose-decomposing bacteria, including sequences related to Anoxybacillus, Leuconostoc, Clostridium, Actinomyces, and Citrobacter. The most abundant bacterial operational taxonomic units (OTUs) in the grass carp intestinal content are those related to feed digestion. In addition, the potential pathogens and probiotics are important members of the intestinal microbiota. Further analyses show that grass carp intestine holds a core microbiota composed of Proteobacteria, Firmicutes, and Actinobacteria. The comparison analyses reveal that the bacterial community in the intestinal contents is most similar to those from the culture water and sediment. However, feed also plays significant influence on the composition of gut microbiota.
MERCURY INTOXICATION IN GRASS CARP (CTENOPHARYNGODON IDELLA)
M. S. Khan, S. A. Khan, Z. I. Chaudhary, M. N. Khan,1 A. Aslam , K. Ashraf2, R. M. Ayyub and M. F. Rai.
Pakistan Veterinary Journal , 2004,
Abstract: The present project was carried out to study the effects of acute and chronic mercury intoxication in Grass carp (Ctenopharyngodon idella). For acute phase experiment, 48 fish were divided into four equal groups (A, B, C and D). Groups B, C and D were given HgCl2 at sublethal dose as 0.4, 0.5 and 0.6 mg/L, respectively, while group A acted as control. Skin, gills and kidneys were isolated from the fish after 48 and 96 hours for pathological studies. For chronic phase, 72 fish were divided into five groups (E, F, G, H and I), containing 12 fish each except group E which contained 24 fish. Groups F, G, H and I were given HgCl2 at sublethal dose of 0.3 mg/L for 4, 8, 12 and 16 days, respectively, while group E acted as control. Skin, gills and kidneys were isolated from each group (F, G, H and I) after 4, 8, 12 and 16 days respectively for pathological studies. During chronic phase in the treatment groups normal histology of epidermis was disturbed with increased number of immature cells. Overall, skin layers were atrophied and withered. Histopathology of gills showed hyperplasia of epithelial cells of gill filaments, fusion of secondary lamellae giving a club shaped appearance of filaments and contraction and sloughing of respiratory epithelium in groups F, G, H and I. Histopathological examination of kidneys also showed a wide range of toxicity lesions and destruction in treatment groups (F, G, H and I). Disintegration and disorganization of cells of both renal and haemopoitic systems including dilatation of capillaries and thickening of basal lumen were observed. Mild to sever tubular epithelial degeneration, karyolysis, dilation and shrinkage of Bowman’s capsule and glomerulus were also observed. In chronic phase experiment, fish showed clinical signs including restlessness, difficult breathing, fin flickering and jerky movements. Suppressed growth rate was also observed in treatment groups (F, G, H and I). During acute phase, after 48 hours, these histopathological lesions were absent in all the groups in all the organs. But after 96 hours, these respective lesions in respective organs were usually of mild, moderate and severe nature in groups B, C and D, respectively. It was concluded that mercury intoxication not only resulted in marked histopathological changes and abnormal clinical signs but also depressed growth rate of fish.
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