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
To test the effectiveness of introducing live zooplankton against direct manuring in ornamental fish ponds upon their survival and production, larvae of koi carp, Cyprinus carpio L., were cultured for 11 weeks in eart...To test the effectiveness of introducing live zooplankton against direct manuring in ornamental fish ponds upon their survival and production, larvae of koi carp, Cyprinus carpio L., were cultured for 11 weeks in earthen ponds maintained according to four management regimes： （1） live zooplankton fed to carp larvae （LF）; （2） direct fertilization with poultry manure （PM）; （3） direct fertilization with cowdung （CD）; and （4） a control treatment （C）. There were three replicates for each treatment. The growth of heterotrophic bacteria and pathogenic microorganisms like Aeromonas sp. and Pseudomonas sp. were also examined in response to pond management. Values of dissolved oxygen were significantly higher （P〈0.05） in the water of LF ponds, compared to other treatments, while the PM and CD treatments recorded were significantly higher （P〈0.05） values of PO4 - P, NH4 - N, NO3 - N, NO2 - N, specific conductivity, alkalinity, and BOD, compared to the LF and C treatments. The percentages of organic carbon and total nitrogen in the bottom sediments were higher in the PM and CD treatments compared to LF （P〈0.05）. Average counts of heterotrophic bacteria in the water of PM and CD ponds were significantly higher than other treatments （P〈0.05）. The development of Aeromonas sp. and Pseudomonas sp. were significantly higher （P〈0.05） in the PM and CD treatments. Weight gain of koi carp stocked in LF was significantly higher （P〈0.05） than that of fish in the other treatments. There was a significant difference in the survival rate of koi carp among the treatments ranging from 67.21% in C to 90.11% in LF. The results suggest that raising koi carp larvae in ponds and feeding them exogenously with zooplankton would support high rates of survival and production through maintenance of better water quality and greater abundance of zooplankton in the system. Significantly lower abundance of Aeromonas sp. and Pseudomonas sp. in the LF treatment considerably lowered any possibility of occurrence of bacterial disease.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
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
Three gram negative bacteria isolated from infected koi carp, Cyprinus carpio L. and screened for sensitivity to thirty antibiotics by the Bauer disc diffusion method. Bacteria evaluated were Aeromonas hydrophila, Pse...Three gram negative bacteria isolated from infected koi carp, Cyprinus carpio L. and screened for sensitivity to thirty antibiotics by the Bauer disc diffusion method. Bacteria evaluated were Aeromonas hydrophila, Pseudomonas aeruginosa and Edwardseilla tarda. A. hydrophila was resistant to ampicilin, cephalothin, sulphatriad, cefpodoxime and cefoxitin. Highest sensitivity was recorded to Colistin methane sulphonate and Ceftriaxone. High resistance was observed for P. aeruginosa in Kanamysin, Cephalothin, Sulphatriad, Colistin methane sulphonate, Tobramysin, Nitro Furantoin, Cefpodoxime, Cefoxitin and Aztreonam. Highest sensitivity recorded with Moxifloxacin. High resistance was observed in Co-trimoxamole and higher sensitivity performed for E. tarda by Tetracyclin, Cephotaxime, Ciprofloxacin, Norfloxacin, Ofloxacin, Moxifloxacin, Gatifloxacin and Levofloxacin. Out of that highest sensitivity recorded with Moxifloxacin High sensitivity was recorded for Moxifloxacin, Ciprofloxacin, Gatifloxacin, Levofloxacin. By means of antibiotic sensitivity test highest resistance was recorded to P. aeruginosa （MAR index-0.3） and least resistant was recorded to E, tarda （MAR index- 0.033）.展开更多
The aim of the present study was to develop a formulated feed for improving skin pigmentation and growth performance of koi carp juveniles. Two experiments were conducted for determination of an optimal dietary astaxa...The aim of the present study was to develop a formulated feed for improving skin pigmentation and growth performance of koi carp juveniles. Two experiments were conducted for determination of an optimal dietary astaxanthin and FM （fish meal） ration for koi carp juveniles. In the first experiment, three isonitrogenous and isoenergetic dietary treatments were designed with three different levels of astaxanthin consisted of 60, 80 and 100 mg/kg and a commercial feed served as a control. For the second experiment, four isonitrogenous and isoenergetic dietary treatments were formulated with graded inclusions of dietary FM replaced from 0% to 60% at 20% increments by SM （soybean meal）, PBM （poultry by-product meal）, and a CD （control diet）. Obtained results demonstrated that skin pigmentation of koi carp juveniles fed the diet containing 80 mg/kg astaxanthin, 36.02% protein, 7.78% lipid, 4.20 Kcal/g GE （gross energy） were more better than those at the diet with lower estaxanthin content and commercial diet. Moreover, the highest growth and feed utilization of fish were observed at this diet with WG （weight gain）, SGR （specific growth rate） and FCR （feed conversion ratio） were 121.80%, 0.95 （%/day） and 1.6, respectively.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.
文摘To test the effectiveness of introducing live zooplankton against direct manuring in ornamental fish ponds upon their survival and production, larvae of koi carp, Cyprinus carpio L., were cultured for 11 weeks in earthen ponds maintained according to four management regimes： （1） live zooplankton fed to carp larvae （LF）; （2） direct fertilization with poultry manure （PM）; （3） direct fertilization with cowdung （CD）; and （4） a control treatment （C）. There were three replicates for each treatment. The growth of heterotrophic bacteria and pathogenic microorganisms like Aeromonas sp. and Pseudomonas sp. were also examined in response to pond management. Values of dissolved oxygen were significantly higher （P〈0.05） in the water of LF ponds, compared to other treatments, while the PM and CD treatments recorded were significantly higher （P〈0.05） values of PO4 - P, NH4 - N, NO3 - N, NO2 - N, specific conductivity, alkalinity, and BOD, compared to the LF and C treatments. The percentages of organic carbon and total nitrogen in the bottom sediments were higher in the PM and CD treatments compared to LF （P〈0.05）. Average counts of heterotrophic bacteria in the water of PM and CD ponds were significantly higher than other treatments （P〈0.05）. The development of Aeromonas sp. and Pseudomonas sp. were significantly higher （P〈0.05） in the PM and CD treatments. Weight gain of koi carp stocked in LF was significantly higher （P〈0.05） than that of fish in the other treatments. There was a significant difference in the survival rate of koi carp among the treatments ranging from 67.21% in C to 90.11% in LF. The results suggest that raising koi carp larvae in ponds and feeding them exogenously with zooplankton would support high rates of survival and production through maintenance of better water quality and greater abundance of zooplankton in the system. Significantly lower abundance of Aeromonas sp. and Pseudomonas sp. in the LF treatment considerably lowered any possibility of occurrence of bacterial disease.
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.
文摘Three gram negative bacteria isolated from infected koi carp, Cyprinus carpio L. and screened for sensitivity to thirty antibiotics by the Bauer disc diffusion method. Bacteria evaluated were Aeromonas hydrophila, Pseudomonas aeruginosa and Edwardseilla tarda. A. hydrophila was resistant to ampicilin, cephalothin, sulphatriad, cefpodoxime and cefoxitin. Highest sensitivity was recorded to Colistin methane sulphonate and Ceftriaxone. High resistance was observed for P. aeruginosa in Kanamysin, Cephalothin, Sulphatriad, Colistin methane sulphonate, Tobramysin, Nitro Furantoin, Cefpodoxime, Cefoxitin and Aztreonam. Highest sensitivity recorded with Moxifloxacin. High resistance was observed in Co-trimoxamole and higher sensitivity performed for E. tarda by Tetracyclin, Cephotaxime, Ciprofloxacin, Norfloxacin, Ofloxacin, Moxifloxacin, Gatifloxacin and Levofloxacin. Out of that highest sensitivity recorded with Moxifloxacin High sensitivity was recorded for Moxifloxacin, Ciprofloxacin, Gatifloxacin, Levofloxacin. By means of antibiotic sensitivity test highest resistance was recorded to P. aeruginosa （MAR index-0.3） and least resistant was recorded to E, tarda （MAR index- 0.033）.
文摘The aim of the present study was to develop a formulated feed for improving skin pigmentation and growth performance of koi carp juveniles. Two experiments were conducted for determination of an optimal dietary astaxanthin and FM （fish meal） ration for koi carp juveniles. In the first experiment, three isonitrogenous and isoenergetic dietary treatments were designed with three different levels of astaxanthin consisted of 60, 80 and 100 mg/kg and a commercial feed served as a control. For the second experiment, four isonitrogenous and isoenergetic dietary treatments were formulated with graded inclusions of dietary FM replaced from 0% to 60% at 20% increments by SM （soybean meal）, PBM （poultry by-product meal）, and a CD （control diet）. Obtained results demonstrated that skin pigmentation of koi carp juveniles fed the diet containing 80 mg/kg astaxanthin, 36.02% protein, 7.78% lipid, 4.20 Kcal/g GE （gross energy） were more better than those at the diet with lower estaxanthin content and commercial diet. Moreover, the highest growth and feed utilization of fish were observed at this diet with WG （weight gain）, SGR （specific growth rate） and FCR （feed conversion ratio） were 121.80%, 0.95 （%/day） and 1.6, respectively.
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%).
