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
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%).
