scholarly journals Artemia sp. as a DNA vaccine vector for common carp Cyprinus carpio larvae

2014 ◽  
Vol 12 (1) ◽  
pp. 54
Author(s):  
Sri Nuryati ◽  
Sekar Sulistyaning Hadiwibowo ◽  
. Alimuddin
Keyword(s):  
Dna Vaccine ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
Herpes Virus ◽  
Oral Vaccine ◽  
Viral Disease ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Glycoprotein Gene ◽  
Herpès Virus

<p class="NoParagraphStyle" align="center"><strong>ABSTRACT</strong></p><p class="NoParagraphStyle" align="center"> </p><p class="NoParagraphStyle">Koi herpes virus (KHV) is one of the most common impetuses for disease on common carp <em>Cyprinus carpio</em>. Generally, viral disease is difficult to cure because virus is intra-cellular parasite, that virus survives, multiplies, and lives only if it on the host cell. Oral vaccine delivery through <em>Artemia</em> sp. is of one alternative way to overcome this problem. This experiment was carried out by analysis DNA vaccine expression encoding of glycoprotein gene (GP-11) on <em>C. carpio</em>. Bacteria containing plasmid Krt-GP-11 as vaccine is served through <em>Artemia </em>sp. as a vector. <em>Artemia</em> sp. was given for one and two times a week to three weeks old common carp. Organs of fish fed by <em>Artemia</em> sp. were analyzed every three days after vaccination. The expression of GP-11 in kidney in each treatment is also observed by the use of RT-PCR method, within ten days after vaccination. The experiment showed that dose of DNA vaccine in whole bacteria could be expressed is 10<sup>6</sup> cfu/mL in a once or twice provisions a week. DNA vaccine could be detected in three organs. RT-PCR analysis also showed that the expression of GP-11 can be detected in all tested organs. In conclusion, <em>Artemia</em> sp. can be used as a vector to carry plasmid GP-11 vaccine for common carp <em>Cyprinus carpio</em> larvae.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: DNA vaccine, KHV, <em>Artemia</em> sp., common carp</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle" align="center"><strong>ABSTRAK</strong></p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Salah satu penyakit pada ikan mas (<em>Cyprinus carpio</em>) yang disebabkan oleh virus adalah <em>koi herpes virus</em> (KHV). Penyakit yang disebabkan oleh virus umumnya sulit untuk disembuhkan karena virus merupakan parasit intraseluler, yaitu virus hanya dapat hidup, bertahan hidup, dan memperbanyak diri di dalam sel inang. Metode pemberian vaksin DNA secara oral melalui <em>Artemia</em> sp. merupakan salah satu alternatif pengobatan yang diharapkan dapat menangani permasalahan penyakit pada ikan yang disebabkan oleh virus. Pada penelitian ini dilakukan uji ekspresi vaksin DNA yang menyandikan glikoprotein 11 (GP-11) pada ikan mas. Bakteri yang mengandung plasmid Krt-GP-11 sebagai vaksin diberikan melalui <em>Artemia</em> sp. sebagai pembawa vaksin. Pemberian <em>Artemia</em> sp. dilakukan satu dan dua kali seminggu pada ikan mas umur tiga minggu. Keberadaan DNA vaksin di usus, ginjal, dan insang dianalisis menggunakan metode PCR. Organ diambil setiap tiga hari setelah pemberian vaksin. Ekspresi gen GP-11 juga diamati pada organ ginjal di setiap perlakuan dengan menggunakan metode RT-PCR, pada sepuluh hari setelah pemberian vaksin. Hasil penelitian menunjukkan bahwa DNA vaksin yang diberikan dengan dosis 10<sup>6</sup> cfu/mL pada perlakuan satu dan dua kali seminggu dapat terdeteksi pada ketiga organ. Hasil RT-PCR menunjukkan bahwa ekspresi GP-11 dapat terdeteksi pada semua organ uji di setiap perlakuan. Dengan demikian <em>Artemia</em> sp. dapat digunakan sebagai vektor pembawa vaksin plasmid GP-11 dengan frekuensi pemberian vaksin untuk larva ikan mas.</p><p class="NoParagraphStyle"> </p><p>Kata kunci: vaksin DNA, KHV, <em>Artemia</em> sp., ikan mas</p>

scholarly journals EFEKTIVITAS EKSTRAK DAUN SAMBILOTO (Andrographis paniculata) BAGI PENGENDALIAN PENYAKIT KOI HERPES VIRUS (KHV) PADA IKAN MAS (Cyprinus carpio)

2007 ◽  
Vol 2 (3) ◽  
pp. 407 ◽  
Author(s):  
Taukhid Taukhid ◽  
Ida Suharni ◽  
Hambali Supriyadi
Keyword(s):  
Common Carp ◽  
Cyprinus Carpio ◽  
Aeromonas Hydrophila ◽  
Herpes Virus ◽  
Andrographis Paniculata ◽  
Control Treatment ◽  
In Vitro Test ◽  
Koi Herpesvirus ◽  
Herpès Virus

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%).

scholarly journals EFEKTIVITAS TRANSFER DAN ANALISIS EKSPRESI GEN IMUNOGENIK TAHAN KOI HERPES VIRUS (KHV) PADA IKAN MAS (Cyprinus carpio)

2014 ◽  
Vol 9 (1) ◽  
pp. 15
Author(s):  
Khairul Syahputra ◽  
Didik Ariyanto ◽  
Erma Primanita Hayuningtyas ◽  
Lamanto Lamanto
Keyword(s):  
Reverse Transcriptase ◽  
Cyprinus Carpio ◽  
Herpes Virus ◽  
Rt Pcr ◽  
Square Wave ◽  
Reverse Transcriptase Pcr ◽  
Herpès Virus

Penelitian transfer gen imunogenik tahan KHV (krt-GP11) pada ikan mas telah dilakukan dengan metode elektroporasi sperma menggunakan konsentrasi DNA yang berbeda. Penelitian ini bertujuan untuk mengetahui konsentrasi DNA optimal yang efektif digunakan dalam transfer gen pada ikan mas. Sperma dielektroporasi menggunakan tipe kejutan square wave dengan voltase 50 V dan jumlah kejutan tiga kali. Konsentrasi DNA yang digunakan adalah 10 μg/mL, 50 μg/mL, dan 100 μg/mL. Deteksi transgen pada sperma, embrio, dan larva dilakukan dengan metode PCR menggunakan primer spesifik untuk gen krt-GP11. Ekspresi transgen pada embrio dan larva dianalisis secara semi-kuantitatif dengan metode reverse transcriptase PCR (RT-PCR). Hasil penelitian menunjukkan bahwa gen krt-GP11 terdeteksi pada sperma, embrio, dan larva. Pemberian konsentrasi DNA 10 μg/mL lebih efektif digunakan dalam transfer gen krt-GP11 pada ikan mas, sedangkan peningkatan konsentrasi DNA yang digunakan tidak memberikan hasil yang berbeda terhadap keberhasilan transfer gen pada ikan mas. Ekspresi gen krt-GP11 yang berhasil diintroduksikan pada ikan mas baru dapat teramati dengan baik pada fase embrio.

scholarly journals Genetic Basis of Cyprinid Herpes Virus-3 Resistance in Common Carp

2011 ◽  
Author(s):  
Lior David ◽  
Yaniv Palti ◽  
Moshe Kotler ◽  
Gideon Hulata ◽  
Eric M. Hallerman
Keyword(s):  
Disease Resistance ◽  
Linkage Map ◽  
Common Carp ◽  
Genetic Linkage ◽  
Herpes Virus ◽  
Genetic Linkage Map ◽  
Viral Disease ◽  
Economic Damage ◽  
Wild Type ◽  
Herpès Virus

The goal of this project was to provide scientific and technical basis for initiating the development of breeding protocols using marker assisted selection for viral disease resistance in common carp. The specific objectives were: 1) Establishing families and characterizing the phenotypic and genetic variation of viral resistance; 2) Measuring the dynamics of immune response and developing a method to measure the long term immune memory; 3) Developing markers and generating a new genetic linkage map, which will enable initial QTL mapping; and, 4) Identifying genetic linkage of markers and candidate genes (like MHC and TLRs) with resistance to CyHV-3. The common carp is an important farmed freshwater fish species in the world. Edible carp is second only to tilapia in Israeli aquaculture production and ornamental carp (koi) is an important product in both the US and Israel. Carp industries worldwide have recently suffered enormous economic damage due to a viral disease caused by Cyprinid herpes virus 3 (CyHV-3). Aside from preventative measures, a sustainable solution to this problem will be to establish a genetic improvement program of the resistance of fish to the pathogen. The aims of the project was to take the necessary first steps towards that. The differences in survival rates after infection with CyHV-3 virus among 20 families from six types of crosses between three carp lines (two commercial lines and one wild-type carp) revealed that the wild-type carp and its crosses had a much-improved survival over the crosses of the commercial lines themselves. These crosses set the starting point for breeding of commercial strains with improved resistance. Resistant fish had lower antibody titer against the virus suggesting that resistance might depend more on the innate immunity. A set of 500 microsateliite markers was developed and the markers are currently being used for generating a genetic linkage map for carp and for identifying disease resistance QTL. Fourteen candidate immune genes, some of which were duplicated, were cloned from the carp and SNP markers were identified in them. The expression of these genes varied between tissues and suggested functional divergence of some duplicated genes. Initial association between CyHV-3 resistance and one of the genes was found when SNP alleles in these genes were tested for their segregation between susceptible and resistant progeny. The results of this project have implications to the development of viral resistant commercial carp strains and effective immunization against this aggressive disease. The genetic and immunological knowledge accumulated in this project will not only promote carp and koi production but will also contribute to a broader understanding of fish immunogenetics.

scholarly journals Construction of a DNA Vaccine Using Glycoprotein Gene and Its Expression Towards Increasing Survival Rate of KHV-Infected Common Carp (Cyprinus carpio)

2012 ◽  
Vol 13 (1) ◽  
pp. 47 ◽  
Author(s):  
Sri Nuryati ◽  
Alimuddin Alimuddin ◽  
Sukenda Sukenda ◽  
Retno Damayanti Soejoedono ◽  
Ayi Santika ◽  
...  
Keyword(s):  
Survival Rate ◽  
Dna Vaccine ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
Vaccine Development ◽  
Fish Muscle ◽  
Negative Control ◽  
Control Fish ◽  
Pcr Analysis ◽  
Fish Group

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.

scholarly journals Koi Herpes Virus (KHV) Disease

EDIS ◽  
1969 ◽  
Vol 2004 (8) ◽  
Author(s):  
Kathleen H. Hartman ◽  
Roy P.E. Yanong ◽  
B. Denise Petty ◽  
Ruth Francis-Floyd ◽  
Allen C. Riggs
Keyword(s):  
Herpes Virus ◽  
Viral Disease ◽  
The United States ◽  
Cooperative Extension Service ◽  
Large Animal ◽  
Information Sheet ◽  
University Of Florida ◽  
Koi Herpesvirus ◽  
Herpès Virus ◽  
Fish Industry

Koi herpes virus (KHV), a viral disease highly contagious to fish, may cause significant morbidity (sickness or disease) and mortality in common carp (Cyprinus carpio) (Hedrick et al., 2000; OATA, 2001). This species is raised as a foodfish in many countries and has been selectively bred for the ornamental fish industry, where it is known as koi. Historically, the first outbreak of KHV was reported in 1998 and confirmed in 1999 in Israel. Since then, other cases have been confirmed in the United States, Europe and Asia (Hedrick et al., 2000; OATA, 2001; Anonymous, 2003). This information sheet is intended to inform veterinarians, biologists, culturists, and hobbyists about KHV. This document is Fact Sheet VM-149, one of a series from the Department of Large Animal Clinical Sciences (College of Veterinary Medicine), Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida. First published: June 2004. VM-149/VM113: Koi Herpesvirus Disease (KHVD) (ufl.edu)

scholarly journals Rapid detection of SARS-CoV-2 infection by multicapillary column coupled ion mobility spectrometry (MCC-IMS) of breath. A proof of concept study

2020 ◽  
Author(s):  
Claus Steppert ◽  
Isabel Steppert ◽  
Gunther Becher ◽  
William Sterlacci ◽  
Thomas Bollinger
Keyword(s):  
Discriminant Analysis ◽  
Ion Mobility ◽  
Influenza A ◽  
Cross Validation ◽  
Viral Disease ◽  
Pcr Analysis ◽  
Proof Of Concept ◽  
Influenza Epidemic ◽  
Rt Pcr ◽  
Non Invasive

AbstractThere is an urgent need for screening patients of having a communicable viral disease to cut infection chains.We could recently demonstrate that MCC-IMS of breath is able to identify Influenza-A infected patients. With decreasing Influenza epidemic and upcoming SARS-CoV-2 infections we extended our study to the analysis of patients with suspected SARS-CoV-2 infections.51 patients, 23m, 28f, aged 64 ± 16 years, were included in this study.Besides RT-PCR analysis of nasopharyngeal swabs all patients underwent MCC-IMS analysis of breath. 16 patients, 7m, 9f, were positive for SARS-CoV-2 by RT-PCR. There was no difference in gender or age according to the groups.Stepwise canonical discriminant analysis could correctly classify the infected and non-infected subjects in 98% by cross-validation. Afterwards we combined the Influenza-A sub study and the SARS-CoV-2-sub study for a total of 75 patients, 34m, 41f, aged 64.8 ± 1.8 years, 14 positive for Influenza-A, 16 positive for SARS-CoV-2, the remaining 44 patients were used as controls. In one patient RT-PCR was highly suspicious of SARS-CoV-2 but inconclusive.There was no imbalance between the groups for age or gender.97.3% of the patients could be correctly classified to the respective group by discriminant analysis. Even the inconclusive patient could be mapped to the SARS-CoV-2 group applying the discrimination function.ConclusionMCC-IMS is able to detect SARS-CoV-2 infection and Influenza-A infection in breath. As this method provides exact, fast non-invasive diagnosis it should be further developed for screening of communicable viral diseases.Study registration: NCT04282135

scholarly journals Diagnóstico etiológico de enfermidades do sistema nervoso central de equinos no Estado de Minas Gerais, Brasil

2015 ◽  
Vol 67 (2) ◽  
pp. 391-399 ◽  
Author(s):  
E.A. Costa ◽  
R. Rosa ◽  
T.S. Oliveira ◽  
R. Furtini ◽  
A.A. Fonseca Júnior ◽  
...  
Keyword(s):  
Herpes Virus ◽  
Minas Gerais ◽  
Rt Pcr ◽  
Saint Louis ◽  
Herpès Virus

O Brasil possui o quarto maior rebanho equino do mundo, e o Estado de Minas Gerais detém o maior número de equinos do país. Portanto, um diagnóstico preciso das doenças neurológicas dos equinos é prioridade no estado. Sendo assim, o objetivo deste estudo foi identificar, utilizando a Reação em Cadeia pela Polimerase (PCR), os agentes infecciosos responsáveis por enfermidades que afetam o sistema nervoso central (SNC) de equinos. De janeiro de 2009 a janeiro de 2011, foi realizado um levantamento dos casos de encefalites e encefalomielites em equinos no Estado de Minas Gerais, utilizando-se amostras de SNC de equinos que morreram com sinais neurológicos. Das 217 amostras de SNC, 47 (21,7%) foram positivas para o vírus da raiva pelo método de imunofluorescência indireta e inoculação em camundongos. Nas 170 amostras negativas para o vírus da raiva, o herpes-vírus equino-1 (EHV-1) foi diagnosticado em 20 (11,8%) e o herpes-vírus suíno-1 (SHV-1), em uma amostra por meio de PCR, e o vírus encefalite de Saint Louis (SLEV), em outra amostra, através de transcrição reversa (RT) e PCR (RT-PCR). Constatou-se que o vírus da raiva é o principal agente causador de encefalite em equinos, apesar do crescente número de casos de encefalomielite associados ao EHV-1 no Estado de Minas Gerais.

Antibody response and resistance of Cyprinus carpio immunized with cyprinid herpes virus 3 (CyHV-3)

Vaccine ◽  
2008 ◽  
Vol 26 (29-30) ◽  
pp. 3750-3756 ◽  
Author(s):  
Ayana Perelberg ◽  
Maya Ilouze ◽  
Moshe Kotler ◽  
Michael Steinitz
Keyword(s):  
Antibody Response ◽  
Cyprinus Carpio ◽  
Herpes Virus ◽  
Herpès Virus
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