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Bergmann, S. M., Y. Jin, K. Franzke, B. Grunow, Q. Wang, and S. Klafack. "Koi herpesvirus (KHV) and KHV disease (KHVD) – a recently updated overview." Journal of Applied Microbiology 129, no. 1 (2020): 98–103. http://dx.doi.org/10.1111/jam.14616.
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Matras, Marek, Jerzy Antychowicz, Jeannette Castric, and Sven M. Bergmann. "CYHV-3 Infection Dynamics in Common Carp (Cyprinus Carpio) – Evaluation of Diagnostic Methods." Bulletin of the Veterinary Institute in Pulawy 56, no. 2 (2012): 127–32. http://dx.doi.org/10.2478/v10213-012-0023-3.
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Abstract The KHV free carp were experimentally infected with CyHV-3 suspension by immersion at temperatures of 18ºC±1 and 25ºC±1. Skin, brain, gills, liver, spleen, kidneys, and leukocytes were investigated for the presence of KHV DNA, and sera for the presence of antibodies against the virus. Similar investigations were performed with non-infected carp, designed as control groups. The results of KHV DNA detection in the infected carp kept at 25ºC ±1 were compared with the results of the virus isolation onto common carp brain cells using leukocyte co-cultivation method and results of specific antibody detection. The appearance of specific antibodies at various time intervals after detecting the clinical symptoms of KHVD in naturally infected carp was additionally studied. At 18°C water temperature, KHV DNA was not detected up to the 6th d post infection (dpi). Then it was found in samples from skin, gill, liver, spleen, and kidneys and much later from the brain. At 25°C water temperature, KHV DNA was already detected on the 3rd dpi in samples from skin and gills, and from the 5th dpi it was found in all examined internal organs, except the brain where it was found at 6th dpi. At 25°C, isolations of CyHV-3 have succeeded between the 7th and 11th dpi. The first specific antibodies were found no sooner than on the 21st dpi. The serological examination performed in naturally infected carp in 15 carp pond farms showed that first specific antiviral antibodies can be detected 2 weeks after the appearance of clinical symptoms of KHVD.
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Klafack, Sandro, Anna-Sophie Fiston-Lavier, Sven Bergmann, et al. "Cyprinid herpesvirus 3 Evolves In Vitro through an Assemblage of Haplotypes that Alternatively Become Dominant or Under-Represented." Viruses 11, no. 8 (2019): 754. http://dx.doi.org/10.3390/v11080754.
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Viruses are able to evolve in vitro by mutations after serial passages in cell cultures, which can lead to either a loss, or an increase, of virulence. Cyprinid herpesvirus 3 (CyHV-3), a 295-kb double-stranded DNA virus, is the etiological agent of the koi herpesvirus disease (KHVD). To assess the influence of serial passages, an isolate of CyHV-3 (KHV-T) was passaged 99 times onto common carp brain (CCB) cells, and virus virulence was evaluated during passages through the experimental infections of common carp. After 78 CCB passages, the isolate was much less virulent than the original form. A comparative genomic analysis of these three forms of KHV-T (P0, P78 and P99) revealed a limited number of variations. The largest one was a deletion of 1363 bp in the predicted ORF150, which was detected in P78, but not in P99. This unexpected finding was confirmed by conventional PCR and digital PCR. The results presented here primarily suggest that, CyHV-3 evolves, at least in vitro, through an assemblage of haplotypes that alternatively become dominant or under-represented.
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Bergmann, S. M., J. Sadowski, M. Kiełpiński, et al. "Susceptibility of koi × crucian carp and koi × goldfish hybrids to koi herpesvirus (KHV) and the development of KHV disease (KHVD)." Journal of Fish Diseases 33, no. 3 (2010): 267–72. http://dx.doi.org/10.1111/j.1365-2761.2009.01127.x.
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Dastagir, Khaled, Rosalia Luketina, Joern Wolfgang Kuhbier, Andreas Ziegler, Peter M. Vogt, and Sören Könneker. "Auswirkung der verspäteten Vorstellung von Handinfektionen beim Handchirurgen – eine retrospektive Single-Center-Studie von 379 Patienten." Handchirurgie · Mikrochirurgie · Plastische Chirurgie 51, no. 01 (2019): 45–48. http://dx.doi.org/10.1055/a-0832-2292.
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Zusammenfassung Ziel Diese Studie untersucht, inwieweit der Vorstellungszeitpunkt bei vorliegender Handinfektion (HI) mit der stationären Liegedauer sowie der Häufigkeit notwendiger Operationen der Patienten korreliert. Patienten und Methoden Retrospektiv wurden die stationären Krankenakten von 379 Patienten mit HI, die zwischen 2007 und 2014 an unserer Klinik (2007–2014) behandelt wurden, statistisch hinsichtlich eines Zusammenhangs zwischen Vorstellungszeitpunkt und Liegedauer sowie der Häufigkeit notwendiger Operationen ausgewertet, wobei auch die Infektionsursache Berücksichtigung fand. Ergebnisse Im Durchschnitt führte eine um mehr als einen Tag verzögerte fachchirurgische Vorstellung zu einer längeren Krankenhausverweildauer von 1,22 Tage (KHVD) (95 %-KI: 1,20–1,25; p < 0,001). Die Notwendigkeit einer operativen Intervention nahm mit jedem Tag Verzögerung um 13,59 % (95 %-KI 4,01 %–25,43 %; p < 0,001) zu. Der verzögerte Beginn einer Antibiotikagabe führte zur Verlängerung der KHVD um den Faktor 1,09 (95 %-KI: 1,03–1,15; p < 0,001), wobei jedoch keine Korrelation bestand zwischen Zeitpunkt der Antibiotikagabe und der Notwendigkeit zur operativen Revision (ja/nein) (p = 0,11). Schlussfolgerung Eine verspätete Vorstellung von Patienten mit Handinfektionen führt zu einer längeren stationären Aufenthaltsdauer sowie zu einer höheren Anzahl an notwendigen Operationen. Eine frühzeitige Vorstellung von HI beim Handchirurgen ist wichtig, um komplizierte Krankheitsverläufe und unnötige Kosten für das Gesundheitssystem zu vermeiden.
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Hammoumi, Saliha, Tatiana Vallaeys, Ayi Santika, et al. "Targeted genomic enrichment and sequencing of CyHV-3 from carp tissues confirms low nucleotide diversity and mixed genotype infections." PeerJ 4 (September 27, 2016): e2516. http://dx.doi.org/10.7717/peerj.2516.
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Koi herpesvirus disease (KHVD) is an emerging disease that causes mass mortality in koi and common carp,Cyprinus carpioL. Its causative agent is Cyprinid herpesvirus 3 (CyHV-3), also known as koi herpesvirus (KHV). Although data on the pathogenesis of this deadly virus is relatively abundant in the literature, still little is known about its genomic diversity and about the molecular mechanisms that lead to such a high virulence. In this context, we developed a new strategy for sequencing full-length CyHV-3 genomes directly from infected fish tissues. Total genomic DNA extracted from carp gill tissue was specifically enriched with CyHV-3 sequences through hybridization to a set of nearly 2 million overlapping probes designed to cover the entire genome length, using KHV-J sequence (GenBank accession numberAP008984) as reference. Applied to 7 CyHV-3 specimens from Poland and Indonesia, this targeted genomic enrichment enabled recovery of the full genomes with >99.9% reference coverage. The enrichment rate was directly correlated to the estimated number of viral copies contained in the DNA extracts used for library preparation, which varied between ∼5000 and ∼2×107. The average sequencing depth was >200 for all samples, thus allowing the search for variants with high confidence. Sequence analyses highlighted a significant proportion of intra-specimen sequence heterogeneity, suggesting the presence of mixed infections in all investigated fish. They also showed that inter-specimen genetic diversity at the genome scale was very low (>99.95% of sequence identity). By enabling full genome comparisons directly from infected fish tissues, this new method will be valuable to trace outbreaks rapidly and at a reasonable cost, and in turn to understand the transmission routes of CyHV-3.
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Taukhid, Taukhid, Desy Sugiani, and Angela Mariana Lusiastuti. "IMPROVEMENT OF NON SPECIFIC IMMUNE SYSTEM ON COMMON CARP (Cyprinus carpio) AGAINST KOI HERPESVIRUS DISEASE (KHVD) BY ADDITION OF ASCORBIC ACID ON FISH DIET: A FIELD SCALE STUDY." Indonesian Aquaculture Journal 5, no. 1 (2010): 45. http://dx.doi.org/10.15578/iaj.5.1.2010.45-51.
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The research with the aim to know the optimal feeding frequency of supplemented ascorbic acid (microencapsulated vitamin C CFC-90) on the dose of 750 mg/kg feed to control Koi Herpesvirus (KHV) disease infecting common carp has been done in field condition. Fish were reared in floating cages with the size of 3.5 m x 3.5 m x 2.0 m and stocking density of 1,250 fish/cage with the size range of ± 10 g/fish. The treatments applied in the research were: (A) daily application, (B) every 3 days application, and (C) without vitamin C addition as the control. Fish test were challenged to KHV infection on the mid cultivation by cohabitation method in the laboratory scale for 2 weeks. Examination on behavior, clinical sign, and mortality of fish test conducted daily. The results showed that the highest survival rate was found on the application of vitamin C every 3 days (60.16%); and followed by every day (52.00%), and the lowest was found on the control group (47.36%).
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Lubis, Dini Sahfitri, and Diah Artati. "DETEKSI KOI HERPES VIRUS DENGAN METODE POLYMERASE CHAIN REACTION." Buletin Teknik Litkayasa Akuakultur 14, no. 2 (2016): 115. http://dx.doi.org/10.15578/blta.14.2.2016.115-118.
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Polymerase chain reaction (PCR) merupakan salah satu metode deteksi cepat koi herpes vrus (KHV). Metode PCR ini sangat sensitif. Sensitivitas tersebut membuatnya dapat digunakan untuk melipatgandakan satu molekul DNA. Konsentrasi dan kualitas DNA dipengaruhi oleh keberhasilan pada saat melakukan ekstraksi DNA. Kegiatan percobaan ini bertujuan untuk mengetahui hasil deteksi virus KHV dengan metode PCR. Keberhasilan analisis PCR sangat dipengaruhi karakteristik DNA genom yang meliputi kemurnian, konsentrasi dan ukuran template. Virus KHV dideteksi menggunakan metode PCR sesuai dengan SNI 7547:2009. Hasil kegiatan deteksi virus KHV menggunakan PCR dari sampel kode I.230–I.286 dapat memvisualisasikan hasil PCR dengan positif KHV dengan ukuran fragmen DNA 290 bp dengan baik. Dapat dilihat dari 57 sampel yang telah diuji memiliki hasil yang berbeda-beda. Hasil uji yang diperoleh yaitu 32 sampel ikan positif terinfeksi KHV dan 25 sampel negatif KHV
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Tu, Chien, Ruey-Yu Chiou, and Meei-Mei Chen. "CLONING, EXPRESSION AND PRELIMINARY ANTIGENICITY ANALYSIS OF STRUCTURAL PROTEINS OF A KOI HERPESVIRUS ISOLATE FROM KOI, CYPRINUS CARPIO IN TAIWAN." Taiwan Veterinary Journal 40, no. 02 (2014): 69–75. http://dx.doi.org/10.1142/s1682648514500097.
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The aim of this study was to clone and express the ORF72 and ORF92 genes of koi herpesvirus (KHV) in a prokaryotic system and to examine the antigenicity of recombinant proteins. Phylogenetic analysis revealed that both ORF72 and ORF92 had 100% homology with KHV-J, and 99% homology with those from KHV-U and KHV-I in nucleotides. This suggests that the KHV isolate in Taiwan is more closely related to the Japanese strain (Asian genotype). In the antigenicity analysis, the crude recombinant ORF72 and ORF92 capsid proteins reacted with the positive sera of the survival fish after a KHV outbreak, indicating that these recombinant capsid proteins might mimic antigens of the wild type KHV to induce an immunological response in the infected host. Our results demonstrated potential for general applicability to serological tests and vaccine development.
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FOMICHEV, ALEXANDER A., and YURI M. MARUSIK. "Five new species of the Acantholycosa-complex (Araneae: Lycosidae) from Mongolia." Zootaxa 4497, no. 2 (2018): 271. http://dx.doi.org/10.11646/zootaxa.4497.2.7.
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Five new species from Mongolia belonging to the Acantholycosa-complex are diagnosed and described: Acantholycosa vahterae sp. n. (♂♀, Govi-Altai Aimag), A. kronestedti sp. n. (♂♀, Govi-Altai Aimag), Mongolicosa azarkinae sp. n. (♂♀, Khovd Aimag), M. cherepanovi sp. n. (♀, Bayan-Ölgii Aimag) and M. ozkutuki sp. n. (♀, Khovd Aimag).
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Evdokimov, A. I., M. G. Zelin, and V. A. Korotkov. "Cyclic hardening of KhVG steel." Metal Science and Heat Treatment 34, no. 9 (1992): 590–92. http://dx.doi.org/10.1007/bf00774608.
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Soto, E., E. Tamez-Trevino, Z. Yazdi, et al. "Non-lethal diagnostic methods for koi herpesvirus in koi Cyprinus carpio." Diseases of Aquatic Organisms 138 (March 26, 2020): 195–205. http://dx.doi.org/10.3354/dao03456.
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Cyprinid herpesvirus 3, also known as koi herpesvirus (KHV), is a viral pathogen responsible for mass mortalities of carp worldwide. In this study, we compared the sensitivity and specificity of ELISA and quantitative PCR (qPCR) methods for the diagnosis of KHV in experimentally infected koi Cyprinus carpio over an 11 mo period. Koi were exposed to KHV at 18 ± 1°C (permissive temperatures for KHV disease) in laboratory-controlled conditions. At 21 d post challenge, the temperature in the system was decreased to <15°C (non-permissive temperature for KHV disease), and fish were monitored for the following 11 mo. At different time points throughout the study, samples of blood and gills were collected from exposed and control koi and subjected to qPCR and ELISA. Survival proportions of 53.3 and 98.8% in exposed and control treatments, respectively, were recorded at the end of the challenge. Traditional receiver-operating characteristic analysis was used to compare the sensitivity of the ELISA and blood and gill qPCR during permissive and non-permissive temperatures. ELISA was superior to qPCR of gills and whole-blood samples in detecting previous exposure to KHV. Similar results were obtained in a second experiment exposing koi to KHV and inducing persistent infection at >30°C (non-permissive temperature for KHV disease). Finally, KHV ELISA specificity was confirmed using cyprinid herpesvirus 1-exposed koi through a period of 3 mo. This study demonstrates that the combination of ELISA and gill qPCR should be recommended in the diagnosis of KHV exposure of suspected carrier-state fish.
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Sungging Pradana, Muhammad, Suwarno Suwarno, and Hari Suprapto. "Deteksi Koi Herpesvirus (KHV) pada Ikan Nila (Oreochromis niloticus) yang Diinfeksi secara Buatan [ Detection of Koi Herpesvirus (KHV) in Nile Tilapia (Oreochromis niloticus) Infected by Artificially Infection ]." Jurnal Ilmiah Perikanan dan Kelautan 7, no. 1 (2019): 39. http://dx.doi.org/10.20473/jipk.v7i1.11230.
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Abstract Koi Herpesvirus (KHV) was formerly known as Cyprinid herpesvirus-3 (CyHV-3) and carp nephritis interstitial and necrosis gill virus (CNGV) is a virus that infects common carp and koi (Cyprinus carpio and C. carpio koi) in farmed and wild population. KHV cause disease at a temperature of 18-25 °C with mortality rate of 80-90 % in fry and adult fish. Currently KHV also detected in tilapia from the results of monitoring in the field. The presence of KHV in tilapia can occur as a result of maintenance in cages adjacent to the infected carp. KHV diagnostic method currently based on case definition and PCR (Polymerase Chain Reaction) detection. The basic concept of PCR is one DNA molecule is used to produce two copies, then four, then eight and so forth through multiplication by polymerase. PCR results sometimes indicated the presence of a faint band caused a low amount of virus, so it is necessary to investigate the presence of KHV DNA in tilapia using different doses of infection. This study aimed to determine the KHV infectivity in nile tilapia were infected by artificially infection and determine dose KHV infection that can infect nile tilapia. The study design used true experimental with with the presentation of descriptive data. Dose of viral infection are 1 ID50, 10 ID50, 100 ID50 and 1000 ID50. The results showed that no clinical symptoms of KHV infected in nile tilapia. The results of electrophoresis of PCR products showed that the mucus of nile tilapia were infected with a 1000 ID50 immersion dose showed thin bands. The same results are also shown in the gill of nile tilapia infected by gill spray at 1000 ID50 dose. Fish were infected by injection, KHV was not detected in mucus, gill, kidney and liver. The results above show nile tilapia cannot be infected by KHV on various treatment
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Syahputra, Khairul, Flandrianto Sih Palimirmo, and Yogi Himawan. "TRANSMISI GEN krt-GP11 DAN PERFORMA KETAHANAN IKAN MAS (Cyprinus carpio) TRANSGENIK F-2 TERHADAP INFEKSI KHV." Jurnal Riset Akuakultur 11, no. 4 (2017): 297. http://dx.doi.org/10.15578/jra.11.4.2016.297-305.
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Pembentukan ikan mas transgenik merupakan salah satu program penelitian di Balai Penelitian Pemuliaan Ikan, Sukamandi dalam rangka menghasilkan varietas unggul ikan mas tahan infeksi KHV (Koi herpesvirus). Pada tahun 2015 telah dilakukan pembentukan ikan mas transgenik tahan KHV generasi F-2. Penelitian ini bertujuan untuk mengevaluasi transmisi gen krt-GP11, ketahanan ikan mas transgenik F-2 terhadap infeksi KHV, keberadaan marka Cyca-DAB1*05 tahan KHV pada populasi ikan mas transgenik F-2. Ikan mas transgenik F-2 dihasilkan dengan memijahkan ikan mas transgenik F-1 jantan dengan betina non-transgenik. Pengujian transmisi transgen dan deteksi marka ketahanan KHV pada transgenik F-2 dilakukan dengan metode PCR menggunakan primer spesifik untuk transgen krt-GP11 dan gen Cyca-DAB1*05. Evaluasi ketahanan ikan mas transgenik F-2 terhadap infeksi KHV dilakukan dengan uji tantang secara kohabitasi. Hasil penelitian menunjukkan bahwa transmisi gen krt-GP11 pada keturunan F-2 memiliki persentase yang relatif rendah yaitu sebesar 0%-2%. Ikan mas transgenik F-2 memiliki ketahanan relatif baik terhadap KHV dengan sintasan uji tantang sebesar 90% dan tidak berbeda nyata dengan ikan mas pembanding atau non-transgenik (P>0,05). Tingginya pesentase keberadaan marka Cyca-DAB1*05 pada populasi transgenik berperan pada ketahanan ikan mas transgenik terhadap infeksi KHV.Creating of transgenic common carp is one of the breeding programs in Research Institute for Fish Breeding for producing a superior strain of common carp resistant to KHV(Koi herpesvirus). Since 2015, the creation of common carp transgenic has been conducted to produce F2 population resistant to KHV. This study was aimed to evaluate the transmission of krt-GP11 gen,the resistantce of F2 transgenic common carp against to KHV infection, and the existence of Cyca-DAB1*05 marker resistant to KHV in F2 transgenic population. F2 transgenic population has been produced by mating F1 transgenic male with non transgenic female. Transgene transmission and the existence of marker resistant to KHV in F2 transgenic population were evaluated by PCR method using specific primer to krt-GP11 gene and Cyca-DAB1*05 gene, respectively. The resistance of F2 transgenic population againstto KHV infection was evaluated by challenge test using cohabitation method. The result showed that transmission of krt-GP11 gene in F2transgenic population was relatively low with percentage of 0-2%. The resistance of F2 transgenic common carp against to KHV was relatively high with survival rate of 90% and was not significantly different from non transgenic (p>0.05). High percentage of transgenic population having Cyca-DAB1*05 marker had a role in resistance of transgenic population against KHV infection.
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Saselah, Jetti Treslah, Reiny A. Tumbol, and Henky Manoppo. "DETERMINASI MOLEKULER KOI HERPES VIRUS (KHV) YANG DIISOLASI DARI IKAN KOI (Cyprinus carpio koi)." JURNAL PERIKANAN DAN KELAUTAN TROPIS 8, no. 2 (2012): 64. http://dx.doi.org/10.35800/jpkt.8.2.2012.408.
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Penelitian ini bertujuan untuk mendeterminasi keberadaan Koi Herpes Virus (KHV) pada ikan koi (Cyprinus carpio koi). Sampel ikan diambil dari Kabupaten Kepulauan Sangihe. Penelitian di lakukan di Laboratorium Biologi Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sam Ratulangi dengan menggunakan metode Polymerase Chain Reaction (PCR). Hasil pemeriksaan PCR terhadap sampel ikan koi mengindikasikan bahwa sampel ikan koi telah terinfeksi oleh virus KHV, yang ditandai dengan munculnya pita DNA pada hasil visualisasi elektroforesis agarosa. Hasil pemeriksaan pada morfologi ternyata sampel ikan yang digunakan menunjukkan gejala-gejala klinis terserang KHV seperti mata pucat, insang berwarna pucat serta produksi lendir yang berlebihan. Beberapa sampel lainnya walaupun secara morfologi belum menunjukan gejala-gejala klinis tetapi melalui pemeriksaan PCR telah terindikasikan terinfeksi KHV. Kata kunci: KHV, PCR, ikan koi The purpose of this study was to determine the presence of Koi Herpes Virus (KHV) in Koi (Cyprinus carpio koi). Fish samples were taken from the Sangihe Archipelago Regency. Laboratory work conducted at the Laboratory of Biology Faculty of Mathematics and Natural Sciences University of Sam Ratulangi using Polymerase Chain Reaction (PCR) method. The samples indicated that the koi fish samples were infected with KHV. It is shown by the appearance of DNA bands on agarose electrophoresis visualization. The morphological examination of the body indicated clinical symptoms of KHV infection, such as pale eyes, pale gills and excessive mucus production. Several other samples, although without clinical symptoms, have shown to be infected as indicated by PCR test. Keywords: KHV, PCR, Koi.
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WAHIDI, BUDI RIANTO, UUN YANUHAR, MOHAMAD MOHAMAD FADJAR, and SRI ANDAYANI. "Pathognomonic features and ultrastructural of Koi Herpesvirus infected Oreochromis niloticus." Biodiversitas Journal of Biological Diversity 20, no. 2 (2019): 497–503. http://dx.doi.org/10.13057/biodiv/d200228.
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Abstract. Wahidi BR, Yanuhar U, Fadjar M, Andayani A. 2019. Pathognomonic features and ultrastructural of Koi Herpesvirus infected Oreochromis niloticus. Biodiversitas 20: 497-503. Koi Herpesvirus (KHV) has caused considerable losses in the cultivation of the Cyprinidae family worldwide. In addition to infecting Cyprinidae, KHV can also infect other freshwater fish. This study reports histopathology and ultrastructural analysis of KHV that infect Nile tilapia. Although there are no specific clinical symptoms, pathognomonic indicating Nile tilapia fish has been infected with KHV have been found, i.e. through the formation of intranuclear inclusion body and cell swelling which essentially experience chromatin margination. In addition, histopathology results indicate changes or damage to other tilapia tissue, i.e. necrosis, hypertrophy, edema, hemorrhage, inflammation, congestion, degeneration, and apoptosis. These results indicate that KHV has been distributed and replicated in tilapia organs. Based on the Transmission Electron Microscopy (TEM) analysis, KHV virions have been detected in the cytoplasm or outside the cells on a size of 150-170 nm.
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Sultan, Makkulau, Stenly Wullur, and Reiny A. Tumbol. "IDENTIFIKASI KOI HERVES VIRUS PADA IKAN MAS Cyprinus carpio DI SULAWESI UTARA TAHUN 2017 DENGAN MENGGUNAKAN TEKNIK PCR DAN qPCR." JURNAL PESISIR DAN LAUT TROPIS 6, no. 2 (2018): 31. http://dx.doi.org/10.35800/jplt.6.2.2018.21521.
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This research aimed to detect the distribution of KHV disease in cultured common carps using conventional PCR and Real Time Quantitative PCR methods in North Sulawesi. The samples were taken from 6 aqua culture centres in North Sulawesi. The results of KHV detection by PCR method showed negative KHV infection because visualization does not form a specific band with the KHV gene that is at 409 bp. Detection of KHV of Ct (Quantification cycle) was greater than the LOD with a confidence level of 95% where Ct LOD is 8.71 for the smallest standard of 1.0x102 copies. Ct sample that was read based on qPCR amplification result which was 14,69-18,80 and the value of Ct NTC (Non Template Control) used as a negative control was 17.52.Penelitian ini bertujuan untuk mengidentifikasi keberadaan penyakit Koi Herves Virus pada ikan mas dengan menggunakan metode PCR dan qPCR di Sulawesi Utara. Sampel uji diambil dari 6 sentral budidaya di Provinsi Sulawesi Utara. Hasil penelitian menunjukkan bahwa dengan metode PCR diperoleh hasil deteksi yang negatif, karena visualisasi tidak terbentuk band spesifik dengan KHV, yaitu di 409 bp. Deteksi KHV dengan metode qPCR didapat hasil infeksi KHV yang negatif dilihat dari nilai rata-rata Ct (Quantification cycle) lebih besar dari LOD dengan tingkat kepercayaan (confident level) 95%. Nilai Ct LOD adalah 8,71 untuk standar terkecil 1,0x102 copies, sedangkan Ct sampel hasil amplifikasi qPCR adalah 14,69-18,80 dan nilai Ct NTC (Non Template Control) yang digunakan sebagai kontrol negatif adalah 17,52.
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Surfianti, Oktarina. "Identifikasi KHV dengan Uji Immunofluorescence dan Immunocytochemistry Berdasarkan Uji Polymerase Chain Reaction Positif KHV pada Ikan Koi (Cyprinus carpio)." Jurnal Biosains Pascasarjana 18, no. 3 (2016): 267. http://dx.doi.org/10.20473/jbp.v18i3.2016.267-282.
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AbstrakKoi Herpesvirus (KHV) menyebabkan penyakit parah dan kematian di segala usia ikan mas dan ikan koi (Cyprinus carpio) dan menyebar dengan cepat di seluruh dunia sebagai penyebab mortalitas yang sangat tinggi dengan perkiraan (80-95%) untuk ikan koi dan ikan mas. Identifikasi KHV ini sudah dilakukan dengan metode Polymerase Chain Reaction (PCR). Tetapi, metode tersebut tidak cocok untuk pekerjaan lapangan karena belum efisien dalam hal waktu dan peralatan. Perlu ada pemeriksaan berbasis imunologis sebagai identifikasi awal (pemeriksaan dini) yang belum banyak dilakukan dan perlu dikembangkan terutama untuk pemeriksaan KHV yaitu metode Immunofluorescence sebagai pemeriksaan imunologis untuk identifikasi adanya antigen atau dengan menggunakan pewarnaan fluorescent dan immunocytochemistry menggunakan metode imuno histo (sito) kimia ensim yang bersifat spesifik dengan pewarnaan oleh zat chromogen guna mengidentifikasi antigen dalam suspensi organ insang dan darah. Tujuan dari penelitian ini adalah agar mampu mengidentifikasi KHV (Koi Herpes Virus) yang menginfeksi ikan koi (Cyprinus carpio L) menggunakan metode aplikasi Immunofluorescence serta Immunocytochemistry berdasarkan pemeriksaan menggunakan uji PCR dengan hasil positif KHV.Hasil dari kedua metode diperoleh positif untuk beberapa sampel berdasarkan positif PCR, ditandai dengan adanya pendaran oleh zat fluorescein berwarna hijau pada antigen KHV dengan pemeriksaan metode Imunofluorescence dan adanya ekspresi zat chromogen pada antigen berwarna coklat kemerahan pada antigen KHV dengan pemeriksaan Imunocytochemistry. Sehingga dapat disimpulkan bahwa kedua metode tersebut mampu mengidentifikasi KHV dan dapat digunakan sebagai uji pemeriksaan dini KHV berbasis imunologis. Kata kunci: Koi Herpesvirus, Ikan koi (Cyprinus carpio), Imunofluorescence, Immunocytochemistry
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Nuryati, Sri, Alimuddin Alimuddin, Sukenda Sukenda, et al. "Construction of a DNA Vaccine Using Glycoprotein Gene and Its Expression Towards Increasing Survival Rate of KHV-Infected Common Carp (Cyprinus carpio)." Jurnal Natur Indonesia 13, no. 1 (2012): 47. http://dx.doi.org/10.31258/jnat.13.1.47-52.
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Deoxyribonucleic acid (DNA) vaccine has recently been developed as an alternative vaccine against virus infection.This study was the first step of DNA vaccine development to protect cyprinids including common carp (Cyprinuscarpio) and fancy koi (Cyprinus carpio) from KHV (koi herpesvirus) infection in Indonesia. One of KHV glycoproteingenes, i.e. glycoprotein (GP) was ligated with Japanese medaka (Oryzias latipes) â-actin promoter to generatepAct/GP as a DNA vaccine. Fourty fish in body weight of 10-15 g/fish were individually injected by pAct/GP intomuscle in different dosage of 2.5 μg, 7.5 μg and 12.5 μg/100 μl phosphate buffer saline. Total RNA was extractedfrom the 12.5 μg of pAct/GP-injected fish muscle at 24, 48 and 67 hours post-injection to analyze GP expression byRT-PCR method. Potential of pAct/GP as DNA vaccine was examined by injecting KHV into the 30-days-vaccinatedfish. Both of possitive and negative control fish group were not vaccinated. Possitive control fish group wereinjected with KHV, but negative control fish group were not. KHV-challenged fish were reared for 1 month, and thedeath fish were calculated daily. Result of RT-PCR analysis showed that GP gene expression were detected at 3 dpost-injection. Expression of GP in the vaccinated fish groups helped to improve their survival rate after challengedby KHV. All of fish without DNA vaccination had dead 17 days after KHV injection. The results demonstrated thatpAct/GP had high potency to be used as a DNA vaccine against KHV infection in cyprinids.
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Suwarno, Faricha Risma Nurani, Hari Suprapto,. "Identifikasi Koi Herpesvirus pada Dosis yang Berbeda dengan Metode Imunohistokimia Streptavidin Biotin pada Ikan Nila (Oreochromis niloticus) [Identification of Koi Herpes Virus At Different Dose with Streptavidin Biotin Methods Immunohistochemistry on Tilapia (Oreochromis niloticus)]." Jurnal Ilmiah Perikanan dan Kelautan 6, no. 2 (2019): 109. http://dx.doi.org/10.20473/jipk.v6i2.11277.
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Abstract KHV is a viral disease in goldfish and Koi Fish (Cyprinus carpio) is a highly contagious, infecting all ages or sizes of fish and aquaculture systems. This disease results in mortality between 80% - 100% of the total fish population, with an incubation period of between 1-7 days. KHV infection triggered by a drop in ambient temperature so it is referred to as a virus that attacks when cold (a cold virus). The disease attacks the temperature range of 18-28 ° C and can cause death. This virus attacks can occur at any age of the fish ranging in size from the seed to the parent. The most prominent clinical symptoms due to KHV infection is the sudden death 1-2 days after infection. Other clinical symptoms are necrosis of the gills, sunken eyes, bleeding at the gills, hemorraghe, excess mucus production in the body, and secondary bacterial infections or parasitic infestations. Streptavidin biotin immunohistochemical method was applied to the study of cells and tissues by staining imunostaining. Technique of determining the existence of (location) antigen (target protein) in tissue using antigen-antibody reaction that begins with histotenik procedure is the procedure of making tissue sections (histology). This method has high sensitivity and fast so that it can be applied to the detection of KHV antigen in tissues of fish Tilapia (Orechromis niloticus). This study aims to determine the presence of KHV antigen in tilapia gills after infection. The method used in this study is the experimental method. Dose of virus titer used was 1 ID50, ID50 10, 100 ID50, ID50 1000. The results showed that the Streptavidin Biotin immunohistochemical examination imunopatologi able to detect KHV virus Ag in the gill tissue of tilapia (Oreochromis niloticus) were infected with different doses. In all treatments showed on all the gills are KHV antigen is indicated by the presence of a golden brown color on the gill epithelium. Advised Streptavidin biotin immunohistochemistry test was applied to the detection of the presence or absence of fish-carp KHV carrier as a routine control program and control, including the prevention and outbreak of KHV in Indonesia, because of KHV were attacked in tilapia was persistent and did not show clinical symptoms
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Koesharyani, Isti, Lila Gardenia, Tatik Mufidah, and Ayi Santka. "APLIKASI KUANTIFIKASI KOI HERPESVIRUS : REAL TIME – QUANTITATIVE POLYMERASE CHAIN REACTION (RT-Q PCR) MENGGUNAKAN SYBR GREEN PADA IKAN MAS (Cyprinus carpio)." Media Akuakultur 12, no. 1 (2017): 45. http://dx.doi.org/10.15578/ma.12.1.2017.45-53.
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Koi Herpes Virus (KHV) di Indonesia sejak tahun 2002 merupakan penyakit mematikan yang menyerang ikan koi Cyprinus carpio koi dan ikan mas Cyprinus carpio carpio, dan sampai saat ini, infeksi KHV dilaporkan sudah menyebar hampir di seluruh dunia. Untuk mengetahui adanya infeksi KHV perlu cara diagnosa yang sangat akurat/sensitif, sehingga keberadaan KHV dapat diketahui secara pasti dengan tingkat sensitivitas yang lebih baik pada ikan budidaya. Tujuan dari penelitian ini adalah untuk mengaplikasikan teknik deteksi dengan real time quantitative polymerase chain reaction (RT- qPCR/qPCR) guna mengetahui adanya infeksi KHV secara kuantitatif pada ikan mas dengan mengetahui kandungan virus (viral load). Sebanyak masing-masing 3 ekor sampel diperoleh dari sentra budidaya ikan mas di Cirata-Jawa Barat, Maninjau-Sumatera Barat, dan Banjarmasin-Kalimantan Selatan. Sampel-sampel tersebut selanjutnya dianalisa keberadaan KHV-nya dengan RT-qPCR menggunakan SYBR Green. Hasil pengujian menunjukkan bahwa jumlah tertinggi (viral load) diperoleh dari ikan mas asal Cirata-3 dengan nilai Threshold Cycle (Ct.) 18,24 atau setara dengan 3,4 x 107 kopi, dan terendah dari ikan mas asal Banjarmasin-3 dengan nilai Ct. 33,39 atau 1,8 x 102 kopi. Dua standar yang digunakan dalam pengujian ini berupa plasmid dengan jumlah kopi 2 x 104 (Ct 27,24) dan 2 x 103 (Ct 30,24) dan kontrol atau Non Template Control (NTC) adalah 3,1 x 10 atau dengan nilai Ct 35,65. Uji aplikasi deteksi KHV dengan metode RT-qPCR ini memberikan hasil yang lebih sensitif, di mana sampel yang tidak terdeteksi dengan metode PCR konvensional dapat dideteksi dan dihitung jumlah kopi DNA (DNA copy). Since 2002, Koi herpesvirus (KHV) in Indonesian has been a malignant diseases, now recognized as a worldwide cause of mortality among populations of koi Cyprinus carpio koi and common carp Cyprinus carpio carpio. To determine the presence of infection is required the KHV diagnosis method with highly accurate and sensitive, so that the existence KHV can be known exactly with high sensitivity level in fish farming.The objective of this study was to develop and evaluate the infection by Real Time Quantitative Polymerase Chain Reaction (RT- qPCR/qPCR). Sample were taken from Carp culture in West Java, West Sumatra, and South Kalimantan. The assay was done by SYBR Green RT-qPCR. The analysis result of KHV in carp revealed that the carp from Cirata-3 had the highest viral load with Ct. value 18.24 equal with 3.4 x 107 copies, and the lowest one was the carp from Banjarmasin-3 at Ct. value 33.39 (1.8 x 102 copies), while two standards plasmid and Non Template Control (NTC) had Ct value of 27.24 (2 x 104copies),30.24 (2 x 103copies), and 35.65 (3.1 x 10 copies), respectively. Application KHV test by q-PCR has more advantages and sensitive than that of conventional PCR, and it can be used to detect and calculate the copy number of DNA.
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Ariyanto, Didik, Erma Primanita Hayuningtyas, and Khairul Syahputra. "KOLEKSI, KARAKTERISASI, DAN SELEKSI PLASMA NUTFAH IKAN MAS (Cyprinus carpio) TAHAN PENYAKIT KOI HERPES VIRUS." Jurnal Riset Akuakultur 9, no. 2 (2014): 215. http://dx.doi.org/10.15578/jra.9.2.2014.215-228.
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Wabah penyakit koi herpes virus (KHV) terjadi sejak tahun 2002 mengakibatkan jumlah produksi ikan mas nasional mengalami penurunan yang cukup signifikan. Salah satu alternatif penanggulangan penyakit KHV yang bisa dilakukan adalah perbaikan genetik untuk membentuk varietas unggul ikan mas tahan KHV. Pembentukan varietas unggul ikan mas tahan KHV dimulai dari kegiatan koleksi, karakterisasi, dan evaluasi plasma nutfah ikan mas. Koleksi plasma nutfah ikan mas dilakukan di beberapa daerah asal ikan mas antara lain di Kabupaten Kuningan, Bandung, Cianjur, dan Pandeglang (Banten). Karakterisasi plasma nutfah ikan mas hasil koleksi dilakukan dengan metode RAPD (Random Amplified Polymorphic DNA) dan evaluasi daya tahan plasma nutfah ikan mas terhadap serangan KHV dilakukan dengan uji tantang secara laboratorium menggunakan metode kohabitasi. Dari hasil kegiatan koleksi diperoleh lima strain ikan mas yang dominan dibudidayakan di Jawa Barat dan Banten, yaitu strain Majalaya (Bandung), Rajadanu (Kuningan), Wildan (Cianjur), Sutisna (Kuningan), dan Sinyonya (Pandeglang). Hasil karakterisasi menunjukkan derajat polimorfisme strain Majalaya, Rajadanu, Wildan, Sutisna, dan Sinyonya secara berturut-turut sebesar 67,71; 83,33; 83,33; 79,17; dan 79,17 dengan heterozigositas sebesar 0,233; 0,274; 0,297; 0,278; dan 0,299. Analisis lanjutan menggunakan program UPGMA mengelompokkan kelima strain tersebut ke dalam tiga kelompok genotipe, yaitu genotipe A (Rajadanu danMajalaya), genotipe B (Sinyonya dan Wildan), serta genotipe C (Sutisna). Hasil uji tantang dengan KHV menunjukkan bahwa strain Rajadanu mempunyai sintasan tertinggi sebesar 40%, diikuti oleh strain Majalaya (36,67%), Sinyonya dan Sutisna (26,67%), serta Wildan (23,33%). Berdasarkan hasil tersebut, strain Rajadanu berpotensi besar dikembangkan sebagai varietas ikan mas tahan KHV.
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Delegnam, Nyamaa. "On mechanisms of increasing interest in Russian language in mongolia (on the example between the Khovd aimag of Mongolia and the Altai Territory of the Russian Federation)." Pedagogy: history, prospects 4, no. 1 (2021): 70–78. http://dx.doi.org/10.17748/2686-9969-2021-4-1-70-78.
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The article examines the features of cross-border cooperation between Russia and Mongolia in order to increase interest in the Russian language and Russian education using the Khovd aimag of Mongolia and the Altai Territory of Russia as examples. The author emphasizes the main directions that contribute to increasing the interest of Mongolian students in the Russian language using mechanisms within the framework of the activities of the Russian Language Center at the Russian Centre of Science and Culture (RCSC) in the Khovd aimag of Mongolia.
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Astuti, Iswari Ratna, Tri Heru Prihadi, Hambali Supriyadi, and Anang Hari Kristanto. "TEKNIK PENGENDALIAN PENYAKIT KHV PADA IKAN MAS (Cyprinus carpio) MELALUI MANIPULASI LINGKUNGAN DALAM SKALA LABORATORIUM." Jurnal Riset Akuakultur 7, no. 3 (2012): 477. http://dx.doi.org/10.15578/jra.7.3.2012.477-484.
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Koi herpes virus merupakan penyakit yang menyebabkan kerugian sangat besar bagi pembudidaya ikan mas. Oleh karena itu, perlu segera ada teknik pengendaliannya. Penelitian ini bertujuan untuk mendapatkan teknik pengendalian penyakit KHV pada benih ikan mas (Cyprinus carpio) melalui manipulasi suhu dan salinitas air. Ikan terinfeksi KHV dipelihara di dalam wadah pemeliharaan volume 45 liter, dengan kepadatan 15 ekor setiap wadah. Ikan uji berukuran 20-25 g/ekor. Penelitian menggunakan Rancangan Acak Kelompok dengan tiga ulangan. Perlakuan terdiri atas kombinasi suhu (26oC-27oC, 29oC-30oC, suhu ruangan) dan salinitas (0, 4, 8, 12 ppt). Hasil penelitian menunjukkan bahwa suhu, salinitas, dan kombinasi suhu dengan salinitas berpengaruh nyata terhadap tingkat kematian ikan karena KHV. Tingkat kematian ikan terinfeksi KHV yang dipelihara pada suhu konstan 26oC-27oC dan 29oC-30oC terbukti lebih rendah dibanding yang dipelihara pada suhu ruang (26oC-30oC). Kombinasi suhu air konstan pada 26oC-30°C dengan salinitas 4–8 ppt adalah yang terbaik untuk pengendalian KHV.
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Costes, B., V. Stalin Raj, B. Michel, et al. "The Major Portal of Entry of Koi Herpesvirus in Cyprinus carpio Is the Skin." Journal of Virology 83, no. 7 (2009): 2819–30. http://dx.doi.org/10.1128/jvi.02305-08.
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ABSTRACT Koi herpesvirus (KHV), recently designated Cyprinid herpesvirus 3, is the causative agent of a lethal disease in koi and common carp. In the present study, we investigated the portal of entry of KHV in carp by using bioluminescence imaging. Taking advantage of the recent cloning of the KHV genome as a bacterial artificial chromosome (BAC), we produced a recombinant plasmid encoding a firefly luciferase (LUC) expression cassette inserted in the intergenic region between open reading frame (ORF) 136 and ORF 137. Two viral strains were then reconstituted from the modified plasmid, the FL BAC 136 LUC excised strain and the FL BAC 136 LUC TK revertant strain, including a disrupted and a wild-type thymidine kinase (TK) locus, respectively. In vitro, the two recombinant strains replicated comparably to the parental FL strain. The FL BAC 136 LUC TK revertant strain was shown in vitro to induce a bioluminescent signal allowing the detection of single positive cells as early as 24 h postinfection, while in vivo, it induced KHV infection in carp that was indistinguishable from that induced by the parental FL strain. To identify the KHV portal of entry, carp were analyzed by bioluminescence imaging at different times postinfection with the FL BAC 136 LUC TK revertant strain. These analyses demonstrated that the skin of the fish covering the fins and also the body is the major portal of entry for KHV in carp. Finally, to further demonstrate the role of the skin as the KHV portal of entry, we constructed an original system, nicknamed “U-tube,” to perform percutaneous infection restricted to the posterior part of the fish. All the data obtained in the present study demonstrate that the skin, and not the gills, is the major portal of entry for KHV in carp.
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Chairunnisa, Sekar Ayu, Sri Nuryati, Alimuddin Alimuddin, Sri Murtini, Ayi Santika, and Dwi Hany Yanti. "EFFICACY OF GP-11 KHV DNA VACCINE IN Cyprinus carpio koi." Indonesian Aquaculture Journal 11, no. 1 (2016): 31. http://dx.doi.org/10.15578/iaj.11.1.2016.31-39.
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Koi herpesvirus (KHV) is one of the major pathogen for koi and common carp which cause high mortality and economic losses for the farmer. The purpose of this study was to determine the efficacy of glycoprotein-11 (GP-11) KHV DNA vaccine and compared to GP-25 KHV DNA vaccine. The vaccine in the form of naked DNA plasmidwas delivered by intramuscularly injection to the 3-month-old koi. The fish were divided into six groups, i.e. unvaccinated group (negative control C- and positive control C+), and vaccinated group (2.5 μg/100 μl of GP-11 (group 1), 7.5 μg/100 μl of GP-11 (group 2), 12.5 μg/100 μl of GP-11 (group 3), and 12.5 μg/100 μl of GP-25 (group 4)). At day 42 post vaccination, all fish of each groups were challenged by injecting KHV titre 10-3 FID50. Number of dead fish was counted everyday after the challenge until 30 days. The results showed that vaccinated fish were had survival rate of 83.33-93.33% (group 2, 3 and 4). It’s show that GP-11 KHV DNA vaccine has high efficacy. As conclusion, GP-11 DNA vaccine could be an alternative DNA vaccine for preventing KHV infection.
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Taukhid, Taukhid, Angela Mariana Lusiastuti, Wulan Andiyani, Rosidah Rosidah, and Sriati Sriati. "INDUKSI KEKEBALAN SPESIFIK PADA IKAN MAS, Cyprinus carpio Linn. TERHADAP INFEKSI KOI HERPESVIRUS (KHV) MELALUI TEKNIK KOHABITASI TERKONTROL." Jurnal Riset Akuakultur 5, no. 2 (2016): 257. http://dx.doi.org/10.15578/jra.5.2.2010.257-276.
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Penelitian dengan tujuan untuk mengetahui masa induksi kekebalan spesifik hingga level protektif terhadap KHV pada populasi ikan mas yang diinfeksi secara buatan melalui teknik pemaparan terkontrol telah dilakukan pada skala laboratorium. Pemaparan terkontrol terhadap ikan positif KHV dilakukan selama 3 hari pada suhu 24oC-26oC, selanjutnya ikan dipindahkan ke wadah volume 300 liter yang diisi ikan uji sebanyak 200 ekor/wadah dan suhu air berkisar antara 31oC-34oC. Perlakuan yang diterapkan adalah periode induksi: (A) periode induksi selama 1 minggu dari akhir masa pemaparan terhadap KHV, (B) periode induksi selama 2 minggu dari akhir masa pemaparan terhadap KHV, (C) periode induksi selama 3 minggu dari akhir masa pemaparan terhadap KHV, (D) periode induksi selama 4 minggu dari akhir masa pemaparan terhadap KHV, dan (E) tanpa periode induksi setelah dilakukan pemaparan terhadap KHV. Pada hari ke-21, seluruh kelompok perlakuan diinfeksi KHV secara buatan dengan teknik kohabitasi. Pengamatan dilakukan terhadap tingkah laku, gejala klinis, dan mortalitas ikan uji yang dilakukan setiap hari hingga akhir percobaan. Rataan persentase sintasan ikan uji tertinggi pada akhir riset diperoleh pada periode induksi selama 3 minggu sebesar 53,75%; diikuti oleh periode induksi selama 2 minggu sebesar 33,75%; selama 1 minggu sebesar 18,75%; selama 4 minggu sebesar 12,5%; dan kelompok kontrol sebesar 7,5%.The research with the aim to study an effective period to develop specific immunity against Koi Herpesvirus on common carp population which was obtained by cohabitation technique had been done in laboratory scale. Cohabitation of KHV infected fish at 24oC-26oC was for 3 days, and then the fish was moved to fiber glass tank at 31oC-34oC to develop specific immunity. The treatments applied in the research were: (A) a week period induction, (B) two weeks period induction, (C) three weeks period induction, (D) four weeks period induction, and (E) without period induction as a control group. Fish test were challenge to KHV infection at the end of each defined period induction by cohabitation method for 2 weeks lasting. Examination on behavior, clinical signs, and mortality of fish test were taken place daily. The results showed that the highest survival rate was found on three weeks period induction (53.75%), and followed by two weeks period induction (33.75%), one week period induction (18.75%), four weeks period induction (12.5%), and the lowest was found on the control (7.5%).
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Matras, Marek, Magdalena Stachnik, Ewa Borzym, Joanna Maj-Paluch, and Michał Reichert. "Potential role of different fish species as vectors of koi herpesvirus (CyHV-3) infection." Journal of Veterinary Research 63, no. 4 (2019): 507–11. http://dx.doi.org/10.2478/jvetres-2019-0069.
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Abstract Introduction Koi herpesvirus (KHV) has infected farmed common carp in Poland clinically and asymptomatically since 2004. The role of non-carp species as vectors of virus transmission is well known except for in the case of KHV. The aim was to better understand this virus’ infection and transmission pathways in common carp, looking at the potential vector role of fishes kept with them. Material and Methods Eight species were experimentally infected with KHV by immersion in a suspension at 20°C ±1 and transferred to a tank after 45 minutes. Specimens were euthanised at intervals up to 56 days post infection (dpi) and tissue was examined for KHV DNA. Surviving infected fishes were introduced at intervals, each time into a separate tank, to naïve common carp for experimental infection. These were observed daily for symptoms, sacrificed along with controls after three months, and dissected to provide tissue samples. Also fish from 14 species collected from a farm with a history of KHV were sampled from 3 to 22 months after disease was confirmed. Organ sections from single fish were collected in a single tube. Results Viral DNA was detected in tench and roach samples up to 49 dpi, but in three-spined stickleback and stone maroko samples only up to 14 dpi. Transmission of KHV to naïve carp occurred after cohabitation. KHV DNA was detected in three fish species three months after the farm outbreak. Conclusion We confirmed that grass and Prussian carp, tench, roach, and brown bullhead can transfer the virus to naïve common carp.
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Asmaul Khusnah, Lilia Widajatiningrum, Ninik Setyorini,. "Kelangsungan Hidup Ikan Koi (Cyprinus carpio koi) Yang Terinfeksi KHV (Koi herpesvirus) [The Survival Of Koi Goldfish (Cyprinus carpio koi) Which Infected By KHV (Koi herpesvirus) ]." Jurnal Ilmiah Perikanan dan Kelautan 1, no. 2 (2019): 135. http://dx.doi.org/10.20473/jipk.v1i2.11679.
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AbstractThe research aim to know the capability of survival life of the infected KHV koi fish that proved with PCR test. The research held from 2003 until 2008, with the methode took gill sample from the same fish every 4 months. Result of PCR showed positive KHV in each analysis during 4.5 years. According to the PCR test for KHV that conducted for 4.5 years showed that the result always positive in every test.
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Antipin, V. S., M. I. Kuzmin, D. Odgerel, L. V. Kousch, and N. V. Sheptyakova. "Comparative geochemical characteristics and genesis of large polystage plutons in the core and periphery of the different age areas of the Mongol-Okhotsk fold belt." Доклады Академии наук 487, no. 4 (2019): 418–23. http://dx.doi.org/10.31857/s0869-56524874418-423.
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The Early Mesozoic Baga-Khentey pluton is the fragment of the Dauria-Khentey batholith, which could have formed due by the mantle plume action on lower horizons of the continental crust within the zone of collisional compression by closing of the Mongol-Okhotsk ocean. The batholith and their peripheral zones possibly formed from the mantle and crustal sources of magma. The Ikh-Narotin-Khid Massif is located on the border of the rifting zones on periphery of the Late Mesozoic area. The petrographic and geochemical affinity of granitoids of the Ikh-Narotin-Khid massif and composition of gneisses from the country rocks might indicate that this was the substratum in formation of palingenic granites of the calc-alkali series. The distinctions in rock composition of the large plutons consist in minor differentiation of the Baga-Khentey Massif rocks probably related to the anatexis conditions and origin of melts in the collisional compression setting. The granites of the Ikh-Narotin-Khid Massif formed in the extension setting favorable for deep differentiation of magmatic melts.
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Sumiati, Tuti, and Agus Sunarto. "ISOLASI KOI HERPESVIRUS (KHV) DARI BEBERAPA ORGAN TARGET DENGAN MENGGUNAKAN KULTUR SEL KT-2." Jurnal Riset Akuakultur 7, no. 1 (2012): 93. http://dx.doi.org/10.15578/jra.7.1.2012.93-100.
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Kasus kematian massal pada ikan mas dan koi (Cyprinus carpio) yang disebabkan oleh koi herpesvirus (KHV) terjadi sejak tahun 2002 dan masih berlangsung hingga sekarang. Pemilihan sampel yang tepat sangat penting untuk mendeteksi dan mengidentifikasi penyakit KHV tersebut. Tujuan dari penelitian ini adalah untuk mengetahui jaringan yang menjadi target infeksi KHV dengan cara isolasi virus menggunakan kultur sel KT-2. Kultur sel diinokulasi dengan ekstrak jaringan organ target (otak, mata, insang, ginjal, limfa, hati, jantung, dan usus, serta gabungan insang, ginjal, dan limfa) dan diinkubasi pada suhu 25oC selama 14 hari. Kerusakan sel terjadi pada kultur sel yang diinokulasi dengan ekstrak dari jaringan insang, ginjal dan gabungan organ insang, ginjal, dan limfa. Uji PCR dari media kultur dan sel yang mengalami CPE menunjukkan bahwa CPE disebabkan oleh KHV.
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Syahputra, Khairul, Didik Ariyanto, and Erma Primanita Hayuningtyas. "KERAGAMAN GENETIK IKAN MAS (Cyprinus carpio) VARIETAS RAJADANU TAHAN KOI HERPESVIRUS GENERASI F0 DAN F1 MENGGUNAKAN TIGA LOKUS MIKROSATELIT." Jurnal Riset Akuakultur 11, no. 1 (2016): 59. http://dx.doi.org/10.15578/jra.11.1.2016.59-66.
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<p>Informasi tentang keragaman genetik sangat dibutuhkan pada program pemuliaan melalui kegiatan seleksi untuk menghasilkan induk unggul, seperti pada pembentukan ikan mas Rajadanu tahan infeksi koi herpes virus (KHV). Penelitian ini bertujuan untuk mengevaluasi variasi genetik ikan mas varietas Rajadanu tahan infeksi KHV generasi F0 dan F1 dengan menggunakan marka molekuler mikrosatelit. Populasi F0 dan F1 dihasilkan dari kegiatan seleksi bersamaan (independent culling) pada karakter pertumbuhan dan ketahanan terhadap KHV. Seleksi karakter pertumbuhan dilakukan dengan metode seleksi individu (mass selection), sedangkan seleksi karakter ketahanan terhadap KHV dilakukan dengan mengidentifikasi individu yang membawa marka MHC II spesifik pada alel Cyca-DAB1*05. Sebanyak sepuluh individu ikan mas dari setiap populasi digunakan untuk analisis variabilitas mikrosatelit menggunakan tiga lokus mikrosatelit (MFW6, MFW7, dan MFW9). Data alel mikrosatelit diolah menggunakan program Microsoft excel dan dianalisis menggunakan program Fstat dan Arlequin. Hasil penelitian menunjukkan bahwa jumlah alel dari tiap lokus pada masingmasing populasi bervariasi, yaitu berkisar antara 2-5 alel. Rata-rata jumlah alel dan rata-rata heterozigositas teramati pada populasi F0 tidak berbeda dengan populasi F1. Rata-rata jumlah alel pada kedua populasi sebesar 3,33 alel dengan rata-rata nilai heterozigositas teramati sebesar 0,47. Inbreeding teridentifikasi pada populasi F0 dan F1, kedua populasi mempunyai tingkat inbreeding yang relatif sama. Populasi ikan mas tahan KHV pada penelitian ini memiliki keragaman genetik yang relatif rendah sehingga diperlukan monitoring variasi genetik dan skema pemijahan yang baik pada kegiatan seleksi selanjutnya untuk menghasilkan ikan mas tahan KHV yang unggul.</p><p>Information on genetic diversity is needed in breeding program through selective breeding to produce superior broodstocks, such as on production of Rajadanu strain of common carp resistant to KHV infection. The aim of this study was to evaluate the genetic variation in F0 and F1 of Rajadanu strain of common carp resistant to KHV infection using microsatellite marker. The F0 and F1 populations have been produced by independent culling method on growth and resistant to KHV characters. Selection on growth character was conducted by mass selection method, while selection on resistant to KHV character was conducted by identification the individual of common carp that carrying MHC II marker specific on CycaDAB1*05 allele. Ten individuals representing each population were analyzed for microsatellite variability using three microsatellite loci (MFW6, MFW7, and MFW9). Microsatellite allele data were analyzed using Microsoft Excel, Fstat, and Arlequin software. The result showed that the number of alleles per loci in each population varied ranging from 2 to 5 alleles. The average number of alleles and the average observed heterozygosity in F0 was similar to that of F1. The average number of alleles in both populations was 3.33, while the average observed heterezygosity was 0.47. The F0 and F1 populations showed inbreeding level; inbreeding level in both populations was relatively similar. Common carp populations in this study had relatively low genetic variation, so that monitoring of genetic variation and good spawning scheme were needed on next selection program to produce a superior common carp resistant to KHV.</p>
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Rouf, Ahmad Beni, Sri Nuryati, Sukenda Sukenda, and Alimuddin Alimuddin. "Efficacy of GP-11 KHV DNA Vaccine in Common Carp (Cyprinus carpio) through Feed by Different Frequency of Administration." Omni-Akuatika 16, no. 1 (2020): 1. http://dx.doi.org/10.20884/1.oa.2020.16.1.768.
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GP-11 KHV DNA vaccine is a vaccine that can be used to induce immunity against the KHV virus (Koi herpesvirus). Vaccination through feed is an alternative way of administering vaccines. The study aimed to examine the effect of giving KHV GP-11 DNA vaccine through feed with different frequencies to KHV infection. The frequency of vaccine administration is GP-11 vaccination once a week; GP-11(1x), GP-11 vaccination twice a week; GP-11(2x), GP-11 vaccination three times a week; GP-11(3x), GP-25 vaccinations three times a week; GP-25(3x), negative control (without KHV test) and positive control (KHV tested). The fish were kept for 28 days after vaccination and then continued with the KHV challenge test for 28 days. The weight of carp ranges from 13.82±2.37 g maintained with a density of 15 fish/aquarium. The results showed that vaccine treatment was able to induce an immune response as indicated by the number of white blood cells, lysozyme activity and post-vaccination antibody titer showed a significant effect compared to controls. Likewise, after the challenge test, supported by IFNγ and IgM gene expression parameters after the challenge test showed the highest value of vaccine treatment rather than control. The efficacy of vaccine was showed by RPS value (%) in each vaccine treatment obtained GP-11(1x) value of 44.7±3.7a, GP-11(2x) of 78.9±18.2b, GP-11(3x) 85.6±12.6b and GP-25(3x) 79.5±18.1b. It was concluded that administering the GP-11 vaccine frequency 2 times a week provides protection as strong as giving a vaccine frequency 3 times a week.Keywords: common carp, DNA vaccine, frequency of administration, koi herpesvirus
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Costes, B., G. Fournier, B. Michel, et al. "Cloning of the Koi Herpesvirus Genome as an Infectious Bacterial Artificial Chromosome Demonstrates That Disruption of the Thymidine Kinase Locus Induces Partial Attenuation in Cyprinus carpio koi." Journal of Virology 82, no. 10 (2008): 4955–64. http://dx.doi.org/10.1128/jvi.00211-08.
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ABSTRACT Koi herpesvirus (KHV) is the causative agent of a lethal disease in koi and common carp. In the present study, we describe the cloning of the KHV genome as a stable and infectious bacterial artificial chromosome (BAC) clone that can be used to produce KHV recombinant strains. This goal was achieved by the insertion of a loxP-flanked BAC cassette into the thymidine kinase (TK) locus. This insertion led to a BAC plasmid that was stably maintained in bacteria and was able to regenerate virions when permissive cells were transfected with the plasmid. Reconstituted virions free of the BAC cassette but carrying a disrupted TK locus (the FL BAC-excised strain) were produced by the transfection of Cre recombinase-expressing cells with the BAC. Similarly, virions with a wild-type revertant TK sequence (the FL BAC revertant strain) were produced by the cotransfection of cells with the BAC and a DNA fragment encoding the wild-type TK sequence. Reconstituted recombinant viruses were compared to the wild-type parental virus in vitro and in vivo. The FL BAC revertant strain and the FL BAC-excised strain replicated comparably to the parental FL strain. The FL BAC revertant strain induced KHV infection in koi carp that was indistinguishable from that induced by the parental strain, while the FL BAC-excised strain exhibited a partially attenuated phenotype. Finally, the usefulness of the KHV BAC for recombination studies was demonstrated by the production of an ORF16-deleted strain by using prokaryotic recombination technology. The availability of the KHV BAC is an important advance that will allow the study of viral genes involved in KHV pathogenesis, as well as the production of attenuated recombinant candidate vaccines.
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Nuswantoro, Soko, Alimuddin, Munti Yuhana, et al. "Efficacy of DNA vaccine encoding koi herpesvirus glycoprotein GP-25in common carp juvenile by immersion." Jurnal Akuakultur Indonesia 11, no. 1 (2013): 76. http://dx.doi.org/10.19027/jai.11.76-85.
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<p class="Default">Koi herpesvirus (KHV) is a herpesvirus that particularly infects and causes mass mortality to koi and common carp. Therefore, the protection of common carp from KHV infection is urgently needed. In this study, we developed an application of DNA vaccine encoding KHV glycoprotein-25 by immersion method to increase survival of common carp against KHV infection. A total of 400 common carp juveniles at 30-day-old were immersed in 1-L water containing 1.3×10<sup>8</sup>CFU/mL of the killed <em>Escherichia coli</em> cells carrying DNA vaccine. Three frequencies and three duration of fish immersion were tested, namely: 1×30 minutes, 1×60 minutes, 1× 90 minutes, 2×90 minutes and 3×90 minutes by interval of 24 hours. Reversetranscription polymerase chain reaction analysis showed that DNA vaccine was successfully expressed in the vaccinated fish. Fish at twenty eight days post vaccination were challenged by injecting 10<sup>-4</sup> mL of KHV per fish. The result showed that vaccination by 1×30 minutes immersion allowed 61% of fish survived, and this was significantly higher (p&lt;0.05) compared to control (without vaccination), but it was similar among vaccination treatments (p&gt;0.05). The relative percent survival of vaccinated fish were also similar among treatments (p&gt;0.05). DNA vaccination has increased fish survival about two fold higher compared to unvaccinated fish control (26.67%). Thus, DNA vaccination was effectively delivered by immersion for 1×30 minutes, and this technique can be useful to level up the resistance of common carp juveniles against KHV infection.</p> <p class="Default">Keywords: DNA vaccine, KHV, glycoprotein, immersion, common carp</p>
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Nuryati, Sri, D. Puspitaningtyas, and D. Wahjuningrum. "Potency of Garlic Extract Against Koi Herpesvirus (KHV) in Common Carp." Jurnal Akuakultur Indonesia 6, no. 2 (2007): 147. http://dx.doi.org/10.19027/jai.6.147-154.
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<p>Prevention of koi herpesvirus (KHV) infection using chemicals or medicines was ineffective way. In this study garlic extract was used to prevent KHV infection. Virus suspension of 0.1 ml and garlic extract of 0.1 ml in different dosage, i.e., 100, 200 and 300 ppt, was injected into common carp body. Hemoglobin concentration, red and white blood cell numbers, and leukocyte number were counted. The results of study showed that administration of 300 ppt of garlic extract could produce higher survival rate (67.5%), good blood parameters and clinical symptoms compared to other treatments.</p> <p>Keywords: garlic, KHV, common carp</p> <p> </p> <p>ABSTRAK</p> <p>Upaya penanggulangan wabah Koi Herpesvirus (KHV) menggunakan bahan-bahan kimia atau obat-obatan adalah tidak efektif. Pada penelitian ini dilakukan pemberian ekstrak bawang putih untuk menanggulangi infeksi KHV. Suspensi virus sebanyak 0,1 ml ditambahkan dengan 0,1 ml ekstrak bawang putih dengan berbagai konsentrasi, yaitu 100, 200 dan 300 ppt, disuntikkan ke dalam tubuh ikan mas. Kadar hemoglobin, jumlah sel darah merah dan sel darah putih jenis dan jumlah leukosit diamati. Hasil penelitian menunjukkan bahwa pemberian ekstrak bawang putih sebanyak 300 ppt menghasilkan kelangsungan hidup (67,5%) yang lebih baik dibandingkan dengan perlakuan lainnya, dan begitu pula dengan gambaran darah serta gejala klinisnya. </p> <p>Kata kunci: bawang putih, KHV, ikan mas</p>
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Ezeoha, Sunday Louis, and Clement Onyeaghala Akubuo. "Influence of palm kernel variables on the yield and quality of oil expressed using an expeller." Research in Agricultural Engineering 67, No. 2 (2021): 92–99. http://dx.doi.org/10.17221/99/2020-rae.
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The objectives of this study were to establish the impact order of the oil-palm kernel processing variables, namely: kernel moisture content (KMC), kernel heating temperature (KHT), kernel heating duration (KHD), and kernel particle size (KPS) on the palm kernel oil (PKO) yield; to develop an empirical model for the PKO yield as influenced by the KMC, KHT and KHD; to investigate the effect of the KMC, KHT and KHD on the PKO quality; and to specify levels of the kernel variables for the maximum PKO yield with minimum variability. The study was undertaken using oil-palm kernels of unidentified variety from Nsukka, Nigeria. The statistical analysis of data was performed with Design-Expert 8P and Minitab 19 Software at P = 0.05. The impact order of the studied kernel variables on the PKO yield, using an expeller (MS-100), was found to be the KPS, KMC, KHD, and KHT. For the maximum PKO yield with minimum variability in the PKO yield, a KMC of 5% (wb), a KHT of 80 °C, a KHD of 10 min, and a KPS of 11 mm and above is recommended.
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Rosenkranz, Daniela, Barbara G. Klupp, Jens P. Teifke, et al. "Identification of envelope protein pORF81 of koi herpesvirus." Journal of General Virology 89, no. 4 (2008): 896–900. http://dx.doi.org/10.1099/vir.0.83565-0.
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Koi herpesvirus (KHV), an emerging pathogen causing mass mortality in koi and common carp, possesses the largest known herpesvirus genome of 295 kbp predicted to encode 156 different proteins. However, none of them has been identified or functionally characterized up to now. In this study, a rabbit antiserum was prepared against a bacterial fusion protein that permitted detection of the predicted type III membrane protein encoded by ORF81 of KHV. In Western blot analyses, the abundant ORF81 gene product of KHV exhibited an apparent mass of 26 kDa and appeared to be non-glycosylated. It could be localized in the cytoplasm of infected cells and in virion envelopes by indirect immunofluorescence and immunoelectron microscopy, respectively. The antiserum was also suitable for the detection of pORF81 in sections of gills, kidneys, hepatopancreas and skin of KHV-infected carp by immunohistochemistry.
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Syahputra, Khairul, Yogi Himawan, Didik Ariyanto, and Flandrianto S. Palim. "PEWARISAN MARKA Cyca-DAB1*05 DAN KERAGAMAN GENETIK IKAN MAS (Cyprinus carpio) STRAIN RAJADANU TAHAN INFEKSI KOI HERPESVIRUS DAN TUMBUH CEPAT." Jurnal Riset Akuakultur 11, no. 2 (2016): 115. http://dx.doi.org/10.15578/jra.11.2.2016.115-123.
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Balai Penelitian Pemuliaan Ikan (BPPI) telah membentuk ikan mas Rajadanu tahan infeksi koi herpesvirus (KHV) dan tumbuh cepat melalui program seleksi. Ikan mas Rajadanu dihasilkan dari kegiatan seleksi bersamaan pada karakter ketahanan terhadap KHV dan pertumbuhan. Seleksi karakter ketahanan terhadap KHV dilakukan dengan menggunakan marka Cyca-DAB1*05 dan seleksi karakter pertumbuhan cepat dilakukan dengan metode seleksi individu. Penelitian ini bertujuan untuk mengevaluasi pewarisan marka Cyca-DAB1*05 dan keragaman genotipe ikan mas varietas Rajadanu tahan infeksi KHV dan tumbuh cepat generasi ketiga (F3). Sebanyak 44 individu ikan mas Rajadanu F3 digunakan pada penelitian ini. Evaluasi marka Cyca-DAB1*05 dilakukan dengan metode PCR menggunakan primer spesifik marka Cyca-DAB1*05. Analisis keragaman genotipe dilakukan menggunakan tiga lokus mikrosatelit (MFW6, MFW7, dan MFW9). Data alel mikrosatelit dianalisis menggunakan program Fstat dan Arlequin. Hasil penelitian menunjukkan bahwa semua individu pada populasi ikan mas Rajadanu F3 membawa marka Cyca-DAB1*05. Ikan mas Rajadanu F3 memiliki keragaman genotipe yang relatif rendah. Nilai rata-rata heterozigositas teramati (0,31) lebih kecil daripada rata-rata heterozigositas harapan (0,46). Nilai positif pada indeks fiksasi (0,32) menunjukkan status inbreeding pada populasi tersebut. Hasil penelitian ini dapat menjadi acuan, baik dalam kegiatan manajemen induk maupun dalam program seleksi selanjutnya dengan tujuan untuk mempertahankan keragaman genetik dan meminimasi tekanan inbreeding ikan mas Rajadanu tahan infeksi KHV dan tumbuh cepat.Research Institute for Fish Breeding (RIFB) has created Rajadanu common carp resistant to koi herpesvirus (KHV) infection and fast growth through selection program. Rajadanu common carp has been produced by independent culling selection on resistant to KHV and fast growth characters. Selection on resistant to KHV and fast growth character was conducted using Cyca-DAB1*05 marker and mass-selection method respectively. The aim of this study was to evaluate the inheritance of Cyca-DAB1*05 as a marker and the genotype variation of Rajadanu common carp resistant to KHV infection and fast growth. A total of 44 fish were used for this study. Evaluation of Cyca-DAB1*05 marker was conducted by PCR method using specific primer to Cyca-DAB1*05 marker. Analysis of genotype variation was conducted using three microsatellite loci (MFW6, MFW7, and MFW9). Microsatellite allele data was analyzed using Fstat and Arlequin software. The result showed that all of F3 Rajadanu common carp were positive carrying Cyca-DAB1*05 marker. Genetic variation of this population was relatively low. The average observed heterozygosity (Ho=0.31) was lower than the average expected heterozygosity (He=0.46). Positive value of fixation index (0.32) showed inbreeding status in this population. The result of this study can be use as reference for broodstock management and next selection program to maintain the genetic diversity and to minimize inbreeding depression level of Rajadanu common carp resistant to KHV infection and fast growth.
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Novita, Hessy, Desy Sugiani, Taukhid Taukhid, and Tuti Sumiati. "DUPLEX POLYMERASE CHAIN REACTION UNTUK DETEKSI SIMULTAN KOI HERPESVIRUS DAN Aeromonas hydrophila PADA IKAN MAS (Cyprinus carpio)." Jurnal Riset Akuakultur 15, no. 1 (2020): 59. http://dx.doi.org/10.15578/jra.15.1.2020.59-67.
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Koi herpesvirus (KHV) dan Aeromonas hydrophila adalah patogen yang dapat mengkoinfeksi ikan mas secara bersamaan. Penelitian ini bertujuan untuk mengembangkan metode duplex polymerase chain reaction (dPCR), deteksi simultan untuk diagnosis KHV dan bakteri Aeromonas hydrophila pada ikan mas. Dua pasang primer yang menargetkan sekuen spesifik SphI dan gen aerolisin, yang sering digunakan untuk mendeteksi KHV dan A. hydrophila dalam uji reaksi tunggal PCR dan menghasilkan target pita PCR 290 bp dan 417 bp. Hasil penelitian menunjukkan bahwa metode duplex PCR dapat mendeteksi ganda infeksi KHV dan A. hydrophila pada ikan mas dan metode ini lebih efektif mendeteksi dua patogen secara bersamaan dalam satu reaksi PCR pada suhu pradenaturasi, 94°C selama dua menit, denaturasi pada suhu 95°C selama satu menit, annealing pada suhu, 55°C selama satu menit, dan 72°C selama satu menit, dengan 30 siklus amplifikasi dan final extention pada suhu 72°C selama lima menit. Metode dPCR untuk deteksi simultan kedua patogen adalah salah satu metode yang dapat diaplikasikan untuk deteksi koinfeksi virus dan bakteri dalam satu reaksi PCR.Koi herpesvirus (KHV) and Aeromonas hydrophila are pathogens that can co-infect common carp. This study aimed to develop a duplex polymerase chain reaction (dPCR) method to detect KHV and Aeromonas hydrophila in common carp simultaneously. Two pairs of primers targeted the specific sequences of SphI and aerolysin genes, often used in detecting KHV and A. hydrophila, in a single PCR reaction test and produced target bands of PCR 290 bp and 417 bp. This proposed method was more effective in simultaneously detecting the two pathogens in one PCR reaction at pre-naturation temperature of 94°C for two minutes, denaturation at 95°C for one minute, annealing at temperature, 55°C for one minute, and 72°C for one minute, with 30 cycles of amplification and final extension at 72°C for five minutes. The findings showed that the duplex PCR method could be used to double detect KHV and A. hydrophila infection in common carp. The duplex PCR method for simultaneous detection of both pathogens is one method that can be applied for the detection of co-infection of viruses and bacteria in a PCR reaction.
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Sumiati, Tuti, and Agus Sunarto. "SUSCEPTIBILITY OF CYPRINID AND NON-CYPRINID FISH CELL LINES TO KOI HERPESVIRUS (KHV)." Indonesian Aquaculture Journal 4, no. 2 (2009): 131. http://dx.doi.org/10.15578/iaj.4.2.2009.131-137.
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Koi herpesvirus (KHV) is an emerging virus that infects koi and common carp (Cyprinus carpio) with mortality up to 95% within 7 days. The disease is rapidly spreading worldwide including to Indonesia. However, it has only been documented in koi and common carp. The aim of this research was to evaluate the susceptibility of fish cell cultures originated from cyprinid and non-cyprinid fish to KHV. Koi Fin (KF-1) and Koi Tail (KT-2) cell lines derived from koi carp and SSN-1 cells originated from fry of striped snakehead were used in this study. The cells were inoculated with tissue extract of KHV-infected koi carp (experiment 1) and virus stock of KHV (experiment 2). The cultures were incubated at 22oC and the onset and type of cytophatic effect (CPE) were observed for 21 days post inoculation. The results of experiment 1 showed that CPE was observed in KT-2 at day 6 post inoculation. In the experiment 2, however CPE was observed in KF-1 and KT-2 cells at day 4 post infection. CPE was not observed in SSN-1 of either experiment 1 or experiment 2. CPE was characterized by extensive vacuolization of the infected cells. Polymerase chain reaction (PCR) assay of cell and tissue culture supernatants confirmed that KF-1 and KT-2 showing CPE were indeed infected with KHV. The results indicated that KF-1 and KT-2 cells were susceptible and SSN-1 was resistant to KHV. The implication of these findings was also discussed in the paper.
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Gilad, Oren, Susan Yun, Mark A. Adkison, et al. "Molecular comparison of isolates of an emerging fish pathogen, koi herpesvirus, and the effect of water temperature on mortality of experimentally infected koi." Journal of General Virology 84, no. 10 (2003): 2661–67. http://dx.doi.org/10.1099/vir.0.19323-0.
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Koi herpesvirus (KHV) has been associated with devastating losses of common carp (Cyprinus carpio carpio) and koi (Cyprinus carpio koi) in North America, Europe, Israel and Asia. A comparison of virion polypeptides and genomic restriction fragments of seven geographically diverse isolates of KHV indicated that with one exception they represented a homogeneous group. A principal environmental factor influencing the onset and severity of disease is water temperature. Optimal growth of KHV in a koi fin cell line occurred at temperatures from 15–25 °C. There was no growth or minimal growth at 4, 10, 30 or 37 °C. Experimental infections of koi with KHV at a water temperature of 23 °C resulted in a cumulative mortality of 95·2 %. Disease progressed rapidly but with lower mortality (89·4–95·2 %) at 28 °C. Mortality (85·0 %) also occurred at 18 °C but not at 13 °C. Shifting virus-exposed fish from 13–23 °C resulted in the rapid onset of mortality.
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Waltzek, Thomas B., Garry O. Kelley, David M. Stone, et al. "Koi herpesvirus represents a third cyprinid herpesvirus (CyHV-3) in the family Herpesviridae." Journal of General Virology 86, no. 6 (2005): 1659–67. http://dx.doi.org/10.1099/vir.0.80982-0.
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The sequences of four complete genes were analysed in order to determine the relatedness of koi herpesvirus (KHV) to three fish viruses in the family Herpesviridae: carp pox herpesvirus (Cyprinid herpesvirus 1, CyHV-1), haematopoietic necrosis herpesvirus of goldfish (Cyprinid herpesvirus 2, CyHV-2) and channel catfish virus (Ictalurid herpesvirus 1, IcHV-1). The genes were predicted to encode a helicase, an intercapsomeric triplex protein, the DNA polymerase and the major capsid protein. The results showed that KHV is related closely to CyHV-1 and CyHV-2, and that the three cyprinid viruses are related, albeit more distantly, to IcHV-1. Twelve KHV isolates from four diverse geographical areas yielded identical sequences for a region of the DNA polymerase gene. These findings, with previously published morphological and biological data, indicate that KHV should join the group of related lower-vertebrate viruses in the family Herpesviridae under the formal designation Cyprinid herpesvirus 3 (CyHV-3).
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Alimuddin, Alimuddin, Mubinun Mubinun, Ayi Santika, Odang Carman, Irvan Faizal, and Komar Sumantadinata. "IDENTIFICATION OF MAJALAYA COMMON CARP STRAINS RESISTANT TO KHV INFECTION USING CYCA-DAB1*05 ALLELE AS THE MARKER." Indonesian Aquaculture Journal 6, no. 2 (2011): 157. http://dx.doi.org/10.15578/iaj.6.2.2011.157-163.
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The Cyca-DAB1*05 allele of major histocompatibility complex class II genes is recently suggested to have a link with the European common carp strain resistant to koi herpesvirus (KHV). In this study, a set of specific primers for Cyca-DAB1*05 was designed and applied as a marker to identify broodstocks of majalaya common carp strain subsequently used as a candidate resistant to KHV infection. From a total of 23 broodstock subjected to PCR analysis, two female and male fish, both having (P) and no Cyca-DAB1*05 (N), were selected and then diallelly mated. Disease resistance of progenies from 10 crosses was determined by a survival analysis in pond rearing and a laboratory challenge-test using cohabitation method. The results have revealed that the average survivals of PxP progenies for pond rearing and KHV challenge test were 86% and 100% higher (P < 0.05) respectively compared to that of NxN fish. Survival rate of PxN/NxP progenies was significantly lower (P < 0.05) than that of PxP fish. Furthermore, PCR analysis showed that almost 91% progenies of PxP crosses seemed to have a KHV resistant gene marker. Thus, this study suggests that the marker is associated with the KHV resistance in majalaya common carp strain, and farming of PxP progenies can be useful to increase common carp production.
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Aoki, Takashi, Ikuo Hirono, Ken Kurokawa, et al. "Genome Sequences of Three Koi Herpesvirus Isolates Representing the Expanding Distribution of an Emerging Disease Threatening Koi and Common Carp Worldwide." Journal of Virology 81, no. 10 (2007): 5058–65. http://dx.doi.org/10.1128/jvi.00146-07.
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ABSTRACT Since the mid-1990s, lethal infections of koi herpesvirus (KHV) have been spreading, threatening the worldwide production of common carp and koi (both Cyprinus carpio). The complete genome sequences of three KHV strains from Japan, the United States, and Israel revealed a 295-kbp genome containing a 22-kbp terminal direct repeat. The finding that 15 KHV genes have clear homologs in the distantly related channel catfish virus (ictalurid herpesvirus 1) confirms the proposed place of KHV in the family Herpesviridae, specifically in the branch with fish and amphibian hosts. KHV thus has the largest genome reported to date for this family. The three strains were interpreted as having arisen from a wild-type parent encoding 156 unique protein-coding genes, 8 of which are duplicated in the terminal repeat. In each strain, four to seven genes from among a set of nine are fragmented by frameshifts likely to render the encoded proteins nonfunctional. Six of the affected genes encode predicted membrane glycoproteins. Frameshifts or other mutations close to the 3′ ends of coding sequences were identified in a further six genes. The conclusion that at least some of these mutations occurred in vivo prompts the hypothesis that loss of gene functions might be associated with emergence of the disease and provides a basis for further investigations into the molecular epidemiology of the virus.
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Pokorova, D., V. Piackova, A. Cizek, et al. "Tests for the presence of koi herpesvirus (KHV) in common carp (Cyprinus carpio carpio) and koi carp (Cyprinus carpio koi) in the Czech Republic." Veterinární Medicína 52, No. 12 (2008): 562–68. http://dx.doi.org/10.17221/1883-vetmed.
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An outbreak of koi herpesvirus (KHV) infection associated with high mortality of common and koi carp was recorded in the USA and Israel in 1998. At present, this disease is viewed as one of the most significant factors that can adversely affect common and koi carp breeds. The disease has spread worldwide including European countries neighbouring with the Czech Republic (CR), i.e. Germany, Poland, and Austria. To monitor the situation in the CR, samples were collected from a total of 138 common and koi carps in seven and eight locations in CR respectively, and were examined between 2005 and 2006. Locations owned by the major producers of common and koi carp were selected with respect of potential occurrence of the KHV virus. No records of increased mortality and morbidity were noticed there. Preferentially carps with non-specific symptoms of disease were sampled, often with isolated skin erosions. To obtain detailed picture about health condition of examined fish the bacteriological and haematological examinations were accomplished. The next part of the examined samples were carp and koi carp for export from professional breeders, imported koi carp and fish from breeds with increased mortality (45 and 21 fish in 2005 and 2006, respectively) The only virological testing was done in this case. The culture and PCR method, according to Gilad et al. (2002) showed negative results for virus KHV in all years. Retrospective investigation by PCR method according to Bercovier et al. (2005) showed positivity in five locations in 2005. In 2006, KHV virus was not detected in any of the selected locations. The aim of our study was to find out the presence/absence of KHV in selected locations and potential correlation of virological, bacteriological and haematological findings. The results of first testing for presence of koi herpesvirus indicates the necessity of regular KHV monitoring in the Czech Republic in the next period.
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Taukhid, Taukhid, Ida Suharni, and Hambali Supriyadi. "EFEKTIVITAS EKSTRAK DAUN SAMBILOTO (Andrographis paniculata) BAGI PENGENDALIAN PENYAKIT KOI HERPES VIRUS (KHV) PADA IKAN MAS (Cyprinus carpio)." Jurnal Riset Akuakultur 2, no. 3 (2007): 407. http://dx.doi.org/10.15578/jra.2.3.2007.407-414.
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Riset ini bertujuan untuk mengetahui efektivitas ekstrak daun sambiloto (Andrographis paniculata) bagi pengendalian penyakit koi herpes virus (KHV) pada ikan mas (Cyprinus carpio). Daun sambiloto dalam bentuk sediaan kering diekstrak melalui perebusan. Pengujian efektivitas antimikrobial dilakukan secara in vitro terhadap bakteri Aeromonas hydrophila sebagai model. Hewan uji yang digunakan adalah ikan mas ukuran 10--15 g/ekor yang secara definitif terinfeksi KHV. Konsentrasi ekstrak daun sambiloto yang diterapkan adalah A (100 mg/L), B (200 mg/L), C (300 mg/L), D (400 mg/L), dan E (tanpa sambiloto sebagai kontrol). Perlakuan dilakukan menggunakan cara perendaman dengan waktu eksposur tidak terbatas. Deteksi KHV pada masingmasing kelompok perlakuan dilakukan setiap minggu dan riset berlangsung selama 3 minggu. Rataan sintasan ikan uji pada kelompok perlakuan adalah A, B, C, D, dan E masing-masing adalah 11,12%, 16,12%, 31,67%, 42,22%, dan 12,78%.The research with the aim to know an efficacy of sambiloto leaf, Andrographis paniculata to control of koi herpesvirus (KHV) on common carp has been conducted in laboratory level. Sambiloto leaf in dry form was extracted by boiling technique. In vitro test of anti microbial properties of sambiloto extract was done against Aeromonas hydrophila isolate as a model. Result of the above research was used as reference for further research. Common carp with the size of 10--15 g/fish, and positively infected by KHV were used as fish test. The treatments applied were A (100 mg/L), B (200 mg/L), C (300 mg/L), D (400 mg/L), and E (without sambiloto extract as a control).Treatment conducted by immersion for indefinite time of exposure. KHV detection of each treatment was done weekly, and research was lasting for 3 weeks. Results of the research showed that mean percentages of survival rate are: A (11.12%), B (16.12%), C (31.67%), D (42.22%), and E (12.78%).
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Syahputra, Khairul, Yogi Himawan, and Didik Ariyanto. "TRANSMISI TRANSGEN GLIKOPROTEIN DAN KETAHANAN IKAN MAS (Cyprinus carpio) TRANSGENIK F1 TERHADAP INFEKSI KOI HERPES VIRUS (KHV)." Jurnal Riset Akuakultur 10, no. 2 (2016): 153. http://dx.doi.org/10.15578/jra.10.2.2015.153-160.
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<p>Ketahanan penyakit merupakan salah satu karakter selain pertumbuhan yang potensial dikembangkan dengan metode transgenesis pada ikan budidaya. Penelitian ini bertujuan untuk mengevaluasi transmisi transgen glikoprotein-GP11 (GP11) dari KHV dan menguji ketahanan ikan mas transgenik F1 terhadap infeksi koi herpes virus (KHV). Empat garis keturunan F1 transgenik (B1, B2, SA1, dan SA2) diproduksi dengan menyilangkan ikan mas jantan F0 yang membawa gen GP11 di sperma dengan betina non-transgenik.<br />Pengujian transmisi transgen dilakukan dengan mendeteksi transgen pada larva dan benih transgenik F1. Deteksi transgen dilakukan dengan metode PCR menggunakan primer spesifik untuk konstruksi gen glikoprotein (krt-GP11). Evaluasi ketahanan terhadap KHV dilakukan dengan uji tantang secara kohabitasi. Hasil penelitian menunjukkan bahwa tidak semua jantan F0 mentransmisikan transgen pada generasi F1. Transmisi transgen pada ikan mas transgenik F1 berkisar antara 0%-3%. Ikan mas transgenik F1 lebih tahan<br />terhadap infeksi KHV dibandingkan non-transgenik. Ikan mas transgenik F1 memiliki sintasan (85,56±7,29%) yang lebih baik dibandingkan dengan ikan mas non-transgenik (71,11±18,99%).</p>
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Al-Rawas, Amer, Hector Sutherland, A. W. Hago, Adnan Basma, and Badar Al-Shihi. "Quantitative Analysis of Clay Minerals Using X-Ray Diffraction Technique. Part 1." Sultan Qaboos University Journal for Science [SQUJS] 3 (December 1, 1998): 31. http://dx.doi.org/10.24200/squjs.vol3iss0pp31-39.
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Qualitative and quantitative analysis of clay minerals are important in the evaluation of the swelling potential of expansive soils A relatively simple method for the quantification of clay minerals of Al-Khod (Northern Oman) using an X-ray diffraction method is described in detail in this paper. The method is based on the additions of known internal standards to the clay sample. The clay investigated in this study contained montmorillonite, palygorskite, illtte and kaolinite. Internal standards of these minerals were mixed with the clay at different proportions and intensities of reflected peak areas were measured. It was found that the peak areas intensities relate linearly with additions of different proportions of standards. From these intensities, the clay minerals present in the clay of Al-khod were estimated.
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Lin, Lisa, Sammi Chen, Duncan S. Russell, et al. "Analysis of stress factors associated with KHV reactivation and pathological effects from KHV reactivation." Virus Research 240 (August 2017): 200–206. http://dx.doi.org/10.1016/j.virusres.2017.08.010.
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