The common carp (Cyprinus carpio) has a large variety of strains. The more popular are the koi (Japanese ornamental carp), which are still bred today to generate creative colors and patterns, giving rise to multiple p...The common carp (Cyprinus carpio) has a large variety of strains. The more popular are the koi (Japanese ornamental carp), which are still bred today to generate creative colors and patterns, giving rise to multiple phenotypes. Since koi are in great demand, there is a challenge to determine the genetics defining their quality. Two methods: 1) direct sequencing of five candidate gene regions, i.e., mitochondrial (cytochrome b, 12S gene and the D-loop) and nuclear (red sensitive opsin and Rag-1) loci, to detect single nucleotide polymorphisms (SNP)s and 2) random amplification of polymorphic DNA (RAPD), were used to differentiate among four koi strains (Kohaku, Sanke, Ghost and Ohgon) and the common carp. Novel SNPs, distinguishing between koi and the common carp, were revealed in cytochrome b, the D-loop and in the red sensitive opsin;one was a missense mutation in cytochrome b at position 15860, in which threonine in the common carp became alanine in all koi strains examined. The Kohaku strain was found to have two alleles in the mitochondrial fragments, forming two different haplotypes (subpo-pulations). These novel SNPs distinguished between koi strains and the common carp, and the RAPD method enabled further differentiation among the four koi strains.展开更多
Koi herpes virus is a new virus found in the aquaculture production of Cryprinus carpiod and common carp in recent years. Currently, virus isolation and identification is still the traditional method for the detection...Koi herpes virus is a new virus found in the aquaculture production of Cryprinus carpiod and common carp in recent years. Currently, virus isolation and identification is still the traditional method for the detection of Koi herpes virus, while molecular biology detection method has become the current developmental di- rection due to its characteristics of more sensitive, specific and rapid. Furthermore, people are still committed to exploring new detection methods for the detection of Koi herpes vires. In this paper, traditional and newly-developed detection methods of Koi herpes vires in recent years were summarized, in order to provide refer- ence for further exploring rapid and accurate diagnostic detection method.展开更多
[ Objective ] This study aimed to establish a rapid and effective quarantine method of Koi herpes virus. [ Method] Primers and corresponding TaqMan probe were designed based on the conserved sequence of Koi herpes vir...[ Objective ] This study aimed to establish a rapid and effective quarantine method of Koi herpes virus. [ Method] Primers and corresponding TaqMan probe were designed based on the conserved sequence of Koi herpes virus (KHV) pol-ymerase gene (Sph) to establish a rapid and effective fluorescence quantitative PCR method for Koi herpes virus detection. The cell cultures were detected by using the established fluorescence quantitative PCR assay, and the results were com- pared with that of conventional PCR. [ Result] The sensitivity of fluorescence quantitative PCR was higher than that of conventional PCR. The minimum copy num- ber that could be detected was 1.6 - 102 copies/p.1. The established method was adopted for sample detection, and a reliable diagnostic result could be obtained within 4 h. [Conclusion] The established method is rapid, sensitive, specific and repeatable, which is conducive to the rapid detection of Koi herpes virus. Key words Koi herpes virus; Fluorescence quantitative PCR; Detection展开更多
A poor environment increases fish’s susceptibility to myxosporean infection that can cause the death of larval fish,especially for koi fish(Cyprinus carpio).This study aimed to determine the effect of probiotics,loca...A poor environment increases fish’s susceptibility to myxosporean infection that can cause the death of larval fish,especially for koi fish(Cyprinus carpio).This study aimed to determine the effect of probiotics,local anti-parasitic drugs(kutuklin),and the chemical compound diflubenzuron treatments on the koi immune response.This study used PCR with specific primer 18S SSU rDNA and DNA sequencing to detect Myxobulus phylogenetic.The treatments were divided into 5 groups:Treatment(A)(healthy koi without treatment),(B)(infected koi without treatment),(C)(infected koi with 0.55 mL/30 L probiotics),(D)(infected koi with 1μL/g of feed kutuklin),and(E)(infected koi with 0.02 mg/5 L dimilin).Myxospore has observed with Scanning Electron Microscopy(SEM)and 4′,6-diamidino-2-phenylindole(DAPI)fluorescence staining.The histological analysis using semi-quantitative scoring methods,and flow cytometry was conducted to analyse the immune response of Cluster of differentiation 4(CD4^(+)),Cluster of differentiation 8(CD8^(+)),Tumor Necrosis Factor-alpha(TNF-α),Interferon gamma(IFN-γ)cells in the gills.Results show that the histological analysis indicated edema,hyperplasia,lamella fusion,congestion,and hypertrophy lesions in infected koi.Treatment with probiotics shows the lowest damage(30.6%).The immune responses of CD4^(+)and CD8^(+)cells to dimilin treatment were 10.54%and 16.86%,respectively.The largest TNF-αand IFN-γresponse were for the kutuklin treatment(29.26%)and probiotics treatment(8.23%).展开更多
文摘The common carp (Cyprinus carpio) has a large variety of strains. The more popular are the koi (Japanese ornamental carp), which are still bred today to generate creative colors and patterns, giving rise to multiple phenotypes. Since koi are in great demand, there is a challenge to determine the genetics defining their quality. Two methods: 1) direct sequencing of five candidate gene regions, i.e., mitochondrial (cytochrome b, 12S gene and the D-loop) and nuclear (red sensitive opsin and Rag-1) loci, to detect single nucleotide polymorphisms (SNP)s and 2) random amplification of polymorphic DNA (RAPD), were used to differentiate among four koi strains (Kohaku, Sanke, Ghost and Ohgon) and the common carp. Novel SNPs, distinguishing between koi and the common carp, were revealed in cytochrome b, the D-loop and in the red sensitive opsin;one was a missense mutation in cytochrome b at position 15860, in which threonine in the common carp became alanine in all koi strains examined. The Kohaku strain was found to have two alleles in the mitochondrial fragments, forming two different haplotypes (subpo-pulations). These novel SNPs distinguished between koi strains and the common carp, and the RAPD method enabled further differentiation among the four koi strains.
基金Supported by Project of Jilin Provincial Science and Technology Commission(20080218)
文摘Koi herpes virus is a new virus found in the aquaculture production of Cryprinus carpiod and common carp in recent years. Currently, virus isolation and identification is still the traditional method for the detection of Koi herpes virus, while molecular biology detection method has become the current developmental di- rection due to its characteristics of more sensitive, specific and rapid. Furthermore, people are still committed to exploring new detection methods for the detection of Koi herpes vires. In this paper, traditional and newly-developed detection methods of Koi herpes vires in recent years were summarized, in order to provide refer- ence for further exploring rapid and accurate diagnostic detection method.
基金Supported by Project of Jilin Province Science and Technology Commission(20080218)
文摘[ Objective ] This study aimed to establish a rapid and effective quarantine method of Koi herpes virus. [ Method] Primers and corresponding TaqMan probe were designed based on the conserved sequence of Koi herpes virus (KHV) pol-ymerase gene (Sph) to establish a rapid and effective fluorescence quantitative PCR method for Koi herpes virus detection. The cell cultures were detected by using the established fluorescence quantitative PCR assay, and the results were com- pared with that of conventional PCR. [ Result] The sensitivity of fluorescence quantitative PCR was higher than that of conventional PCR. The minimum copy num- ber that could be detected was 1.6 - 102 copies/p.1. The established method was adopted for sample detection, and a reliable diagnostic result could be obtained within 4 h. [Conclusion] The established method is rapid, sensitive, specific and repeatable, which is conducive to the rapid detection of Koi herpes virus. Key words Koi herpes virus; Fluorescence quantitative PCR; Detection
基金The authors would like to express their gratitudes to the Institute for Research and Community Service,Universitas Brawijaya,Indonesia through the“Doktor Mengabdi 2018"[grant DIPA number:DIPA-042.01.2.400919,2018].
文摘A poor environment increases fish’s susceptibility to myxosporean infection that can cause the death of larval fish,especially for koi fish(Cyprinus carpio).This study aimed to determine the effect of probiotics,local anti-parasitic drugs(kutuklin),and the chemical compound diflubenzuron treatments on the koi immune response.This study used PCR with specific primer 18S SSU rDNA and DNA sequencing to detect Myxobulus phylogenetic.The treatments were divided into 5 groups:Treatment(A)(healthy koi without treatment),(B)(infected koi without treatment),(C)(infected koi with 0.55 mL/30 L probiotics),(D)(infected koi with 1μL/g of feed kutuklin),and(E)(infected koi with 0.02 mg/5 L dimilin).Myxospore has observed with Scanning Electron Microscopy(SEM)and 4′,6-diamidino-2-phenylindole(DAPI)fluorescence staining.The histological analysis using semi-quantitative scoring methods,and flow cytometry was conducted to analyse the immune response of Cluster of differentiation 4(CD4^(+)),Cluster of differentiation 8(CD8^(+)),Tumor Necrosis Factor-alpha(TNF-α),Interferon gamma(IFN-γ)cells in the gills.Results show that the histological analysis indicated edema,hyperplasia,lamella fusion,congestion,and hypertrophy lesions in infected koi.Treatment with probiotics shows the lowest damage(30.6%).The immune responses of CD4^(+)and CD8^(+)cells to dimilin treatment were 10.54%and 16.86%,respectively.The largest TNF-αand IFN-γresponse were for the kutuklin treatment(29.26%)and probiotics treatment(8.23%).