scholarly journals Frequency and persistency of DNA vaccine encoding GP25 by oral on common carp

2015 ◽  
Vol 12 (2) ◽  
pp. 150
Author(s):  
Sri Nuryati ◽  
, Yuliyanti ◽  
, Alimuddin
Keyword(s):  
Escherichia Coli ◽  
Dna Vaccine ◽  
Common Carp ◽  
Oral Delivery ◽  
Culture Medium ◽  
Dose Frequency ◽  
Viral Pathogen ◽  
Koi Herpesvirus ◽  
Pcr Method ◽  
Almost All

<p class="NoParagraphStyle" align="center"><strong>ABSTRACT</strong></p><p class="NoParagraphStyle" align="center"> </p><p class="NoParagraphStyle">Koi herpesvirus (KHV) is a major viral pathogen that infects common carp and koi. KHV disease outbreak is happened in almost all centre of common carp culture in Indonesia and caused mass mortality. Deoxyribonucleic acid (DNA) vaccination method is one of ways to cope with KHV infection. Vaccines were commonly given by injection. The aim of this research was to get frequency and persistency of DNA vaccine encoding GP25 given by oral delivery method in common carp. This research would like to determine dose, frequency of vaccination, persistency of DNA vaccine and culture medium for the bacterial host. DNA vaccine persistency test was done by using polymerase chain reaction (PCR) method with the specific primer for GP25 gene. The results showed that level of DNA vaccine that could be detected in feed was 7.56 ng (equal to 1.598×10<sup>10</sup> copies). Efficient culture medium for <em>Escherichia coli</em> DH5α carrying DNA vaccine was LB triptone. Feeding fish with diet supplemented with 1 mL <em>E. coli</em> DH5α containing DNA vaccine for each fish and two times a week allowed persistence of DNA vaccine in kindney and spleen.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: common carp, KHV, DNA vaccine, GP25, persistance</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">Koi herpesvirus (KHV) adalah virus patogen utama yang menginfeksi ikan mas dan ikan koi. Wabah penyakit KHV terjadi di hampir semua sentra budidaya ikan mas di Indonesia dan menyebabkan kematian massal ikan. Metode vaksinasi DNA merupakan salah satu cara yang dapat dilakukan untuk menanggulangi serangan KHV. Pemberian vaksin umumnya dilakukan dengan cara injeksi. Tujuan penelitian ini adalah untuk menguji frekuensi dan persistensi vaksin DNA GP25 antivirus KHV yang diberikan melalui oral pada ikan mas. Pada penelitian ini dilakukan uji dosis, frekuensi pemberian vaksin, persistensi vaksin DNA, dan media kultur bakteri inang. Persistensi vaksin DNA dianalisis menggunakan metode PCR dengan primer spesifik gen GP25. Hasil penelitian menunjukkan bahwa dosis vaksin DNA yang dapat terdeteksi dalam pakan adalah 7,56 ng (setara dengan 1,598×10<sup>10</sup> copy). Media kultur yang efisien bagi bakteri Escherichia coli DH5α pembawa vaksin DNA adalah LB tripton. Pemberian pakan bervaksin berupa bakteri konsentrasi 1 mL/ekor ikan dengan frekuensi dua kali seminggu menghasilkan persistensi DNA GP25 di ginjal dan limpa.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Kata kunci: ikan mas, KHV, vaksin DNA, GP25, persistensi</p><p> </p>

scholarly journals TRANSMISI GEN krt-GP11 DAN PERFORMA KETAHANAN IKAN MAS (Cyprinus carpio) TRANSGENIK F-2 TERHADAP INFEKSI KHV

2017 ◽  
Vol 11 (4) ◽  
pp. 297
Author(s):  
Khairul Syahputra ◽  
Flandrianto Sih Palimirmo ◽  
Yogi Himawan
Keyword(s):  
Common Carp ◽  
Challenge Test ◽  
Specific Primer ◽  
Breeding Programs ◽  
Koi Herpesvirus ◽  
F2 Population ◽  
Fish Breeding ◽  
Pcr Method ◽  
Transgenic Male ◽  
Transgene Transmission

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.

scholarly journals Efficacy of GP-11 KHV DNA Vaccine in Common Carp (Cyprinus carpio) through Feed by Different Frequency of Administration

Omni-Akuatika ◽  
2020 ◽  
Vol 16 (1) ◽  
pp. 1
Author(s):  
Ahmad Beni Rouf ◽  
Sri Nuryati ◽  
Sukenda Sukenda ◽  
Alimuddin Alimuddin
Keyword(s):  
Dna Vaccine ◽  
Common Carp ◽  
White Blood Cells ◽  
Challenge Test ◽  
Positive Control ◽  
Negative Control ◽  
Lysozyme Activity ◽  
Post Vaccination ◽  
Koi Herpesvirus ◽  
Vaccine Treatment

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

scholarly journals Intra-muscular and oral vaccination using a Koi Herpesvirus ORF25 DNA vaccine does not confer protection in common carp (Cyprinus carpio L.)

2019 ◽  
Vol 85 ◽  
pp. 90-98 ◽  
Author(s):  
Carmen W.E. Embregts ◽  
Roni Tadmor-Levi ◽  
Tomáš Veselý ◽  
Dagmar Pokorová ◽  
Lior David ◽  
...  
Keyword(s):  
Dna Vaccine ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
Oral Vaccination ◽  
Koi Herpesvirus ◽  
Common Carp Cyprinus Carpio ◽  
Carp Cyprinus Carpio

scholarly journals Potential transmission test of GP25 vaccine in normal flora bacteria of common carp culture media

2015 ◽  
Vol 14 (1) ◽  
pp. 90
Author(s):  
Sri Nuryati ◽  
, Alimuddin ◽  
Ayu Dhita Juliadiningtyas
Keyword(s):  
Dna Vaccine ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
Dna Vaccines ◽  
Culture Media ◽  
Pond Water ◽  
Vaccine Safety ◽  
Normal Flora ◽  
Koi Herpesvirus ◽  
Transmission Test

<p class="NoParagraphStyle" align="center"><strong>ABSTRACT</strong><strong></strong></p><p class="NoParagraphStyle" align="center"><strong> </strong></p><p class="NoParagraphStyle">Koi herpesvirus (KHV) is a virus that infects common carp and koi <em>Cyprinus carpio</em>. KHV attacks most stadia of common carp and koi and causes mortality up to 80–95% of the population. One way to prevent the spread of KHV is  by applying DNA vaccine. This research was conducted to test potential transmission of DNA vaccine encoding glycoprotein 25 (vaccine GP25) toward normal flora bacteria of media cultivation of common carp. Bacteria was isolated from pond water of common carp and tested for their sensitivity to ampicillin. Research was performed by adding vaccine GP25 to ampicillin-sensitive bacteria at a dose of 12.5 µg/100 µL and incubated at 28 °C for 30, 60, 180, and 300 minutes then plated on media containing ampicillin. The grown bacteria cells were tested for the existence of plasmid bearing gen GP25 through bacteria colony cracking. The results of this research showed that there was no bacteria contained plasmid bearing gen GP25.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: DNA vaccines, common carp, vaccine safety</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"><em>Koi herpesvirus</em> (KHV) merupakan virus yang menginfeksi ikan mas dan koi <em>Cyprinus carpio</em>. KHV menyerang hampir semua stadia ikan mas dan koi dan menyebabkan kematian hingga 80–95% dari populasi. Salah satu cara penanggulangan penyebaran KHV adalah dengan penggunaan vaksin DNA. Penelitian ini dilakukan untuk menguji potensi transmisi vaksin DNA glikoprotein 25 (vaksin GP25) pada bakteri flora normal pada media budidaya ikan mas. Bakteri diisolasi dari kolam budidaya ikan mas dan diuji sensitivitasnya terhadap antibiotik ampisilin. Penelitian dilakukan dengan menambahkan vaksin GP25 ke larutan bakteri sensitif ampisilin dengan dosis 12,5 µg/100 µL dan diinkubasi pada suhu 28 °C selama 30, 60, 180, dan 300 menit kemudian disebar pada media mengandung antibiotik ampisilin. Sel bakteri yang tumbuh diuji untuk mendeteksi keberadaan plasmid pembawa gen GP25 dengan menggunakan metode seleksi koloni bakteri (<em>colony cracking</em>). Hasil penelitian ini menunjukkan bahwa tidak ada bakteri yang mengandung plasmid pembawa gen GP25.</p><p class="NoParagraphStyle"> </p><p>Kata kunci: keamanan vaksin, ikan mas, vaksin DNA</p>

scholarly journals Efficacy of DNA vaccine encoding koi herpesvirus glycoprotein GP-25in common carp juvenile by immersion

2013 ◽  
Vol 11 (1) ◽  
pp. 76 ◽  
Author(s):  
Soko Nuswantoro ◽  
. Alimuddin ◽  
Munti Yuhana ◽  
Ayi Santika ◽  
Sri Nuryati ◽  
...  
Keyword(s):  
Dna Vaccine ◽  
Common Carp ◽  
Polymerase Chain Reaction Analysis ◽  
Dna Vaccination ◽  
Chain Reaction ◽  
Immersion Method ◽  
Post Vaccination ◽  
Koi Herpesvirus ◽  
Polymerase Chain ◽  
Fish Survival

<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>

scholarly journals DUPLEX POLYMERASE CHAIN REACTION UNTUK DETEKSI SIMULTAN KOI HERPESVIRUS DAN Aeromonas hydrophila PADA IKAN MAS (Cyprinus carpio)

2020 ◽  
Vol 15 (1) ◽  
pp. 59
Author(s):  
Hessy Novita ◽  
Desy Sugiani ◽  
Taukhid Taukhid ◽  
Tuti Sumiati
Keyword(s):  
Polymerase Chain Reaction ◽  
Common Carp ◽  
Aeromonas Hydrophila ◽  
Simultaneous Detection ◽  
Duplex Pcr ◽  
Chain Reaction ◽  
Koi Herpesvirus ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Duplex Polymerase Chain Reaction

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.

scholarly journals Hematology of common carp following DNA vaccination and koi herpesvirus challenge test

2010 ◽  
Vol 9 (1) ◽  
pp. 9 ◽  
Author(s):  
Sri Nuryati ◽  
N.A. Maswan ◽  
. Alimuddin ◽  
. Sukenda ◽  
K. Sumantadinata ◽  
...  
Keyword(s):  
Survival Rate ◽  
Dna Vaccine ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
Lethal Dose ◽  
Challenge Test ◽  
Phagocytic Index ◽  
Viral Glycoprotein ◽  
Post Vaccination ◽  
Koi Herpesvirus

<p>The study was aimed to determine the effectiveness of DNA vaccine doses on hematological aspect which represent immune response and its influence on common carp survival rate. DNA vaccines encoding the viral glycoprotein of  koi herpesvirus (KHV) have been proved to highly protect the fish under laboratory condition.  A dose of 12.5 µg/100 µl vaccine had resulted in a survival rate of 96.67 % during 30 days after challenge test with a lethal dose of KHV. Fish vaccinated using lower doses, i.e. 2.5 and 7.5 µg/100µl showed 100% mortality after 15 and 19 days challenge test respectively, whereas non vaccinated fish as a control showed 100% mortality after 17 days challenge test.  Total leucocytes of the vaccinated fish were higher than control until 42 days post vaccination, but declined afterward.  Phagocytic index of the vaccinated fish using 12.5 µg/100 µl was declined after 49 days post vaccination or 7 days post challenge test.</p> <p>Key words: DNA vaccine, Koi herpesvirus (KHV), leucocyte, phagocytic index,<em> Cyprinus carpio</em></p> <p> </p> <p>ABSTRAK</p> <p>Penelitian ini bertujuan untuk menguji pengaruh vaksinasi menggunakan vaksin DNA dengan dosis berbeda terhadap gambaran darah ikan sebagai respresentasi tanggap kebal ikan mas serta pengaruhnya terhadap tingkat kelangsungan hidup ikan mas. Vaksin DNA penyandi glikoprotein koi herpesvirus (KHV) dapat memberikan proteksi yang tinggi pada percobaan skala laboratorium.  Vaksinasi dengan dosis 12,5 µg/100µl dapat mempertahankan kelangsungan hidup sebesar 96,67% selama satu bulan setelah uji tantang dengan virus KHV menggunakan dosis letal.  Ikan yang divaksin dengan dosis yang lebih rendah yaitu 2,5 dan 7,5 µg/100µl mengalami kematian total berturut-turut setelah 15 dan  19 hari uji tantang, sedangkan ikan kontrol yang tidak divaksin mengalami kematian total setelah 17 hari uji tantang.  Jumlah leukosit total ikan yang divaksinasi lebih tinggi dibanding dengan kontrol sampai hari ke-42, setelah itu mengalami penurunan.  Indeks fagositosis ikan yang divaksin dengan dosis 12,5 µg/100µl mengalami penurunan setelah hari ke-49 atau 7 hari setelah uji tantang.</p> <p>Kata kunci: Vaksin DNA, Koi herpesvirus (KHV), leukosit, indeks fagositosis,<em> Cyprinus carpio</em></p>

scholarly journals Koi Herpesvirus Disease (KHVD)

EDIS ◽  
2013 ◽  
Vol 2013 (5) ◽  
Author(s):  
Kathleen H. Hartman ◽  
Roy P. Yanong ◽  
Deborah B. Pouder ◽  
Barbara Denise Petty ◽  
Ruth Francis-Floyd ◽  
...  
Keyword(s):  
United Kingdom ◽  
Common Carp ◽  
Ornamental Fish ◽  
Significant Morbidity ◽  
Fact Sheet ◽  
Koi Herpesvirus ◽  
The United Kingdom ◽  
Aquatic Sciences ◽  
Almost All ◽  
Fish Industry

Koi herpesvirus (KHV) is a highly contagious virus that causes significant morbidity and mortality in common carp varieties. Common carp is raised as a foodfish in many countries and has also been selectively bred for the ornamental fish industry where it is known as koi. The first recognized case of KHV occurred in the United Kingdom in 1996. Since then other cases have been confirmed in almost all countries that culture koi and/or common carp with the exception of Australia. This 9-page fact sheet is intended to inform veterinarians, biologists, fish producers and hobbyists about KHV disease. Written by Kathleen H. Hartman, Roy P.E. Yanong, Deborah B. Pouder, B. Denise Petty, Ruth Francis-Floyd, Allen C. Riggs, and Thomas B. Waltzek, and published by the UF Department of Fisheries and Aquatic Sciences, April 2013. http://edis.ifas.ufl.edu/vm113

scholarly journals RT-PCR analysis of Deformed wing virus in honeybees (Apis mellifera) and mites (Varroa destructor)

2005 ◽  
Vol 86 (12) ◽  
pp. 3419-3424 ◽  
Author(s):  
Constanze Yue ◽  
Elke Genersch
Keyword(s):  
Varroa Destructor ◽  
Pcr Analysis ◽  
Body Parts ◽  
Rt Pcr ◽  
Larval Food ◽  
Viral Pathogen ◽  
Deformed Wing Virus ◽  
Significant Difference ◽  
Almost All ◽  
Viral Sequences

Deformed wing virus (DWV) is a honeybee viral pathogen either persisting as an inapparent infection or resulting in wing deformity. The occurrence of deformity is associated with the transmission of DWV through Varroa destructor during pupal stages. Such infections with DWV add to the pathology of V. destructor and play a major role in colony collapse in the course of varroosis. Using a recently developed RT-PCR protocol for the detection of DWV, individual bees and mites originating from hives differing in Varroa infestation levels and the occurrence of crippled bees were analysed. It was found that 100 % of both crippled and asymptomatic bees were positive for DWV. However, a significant difference in the spatial distribution of DWV between asymptomatic and crippled bees could be demonstrated: when analysing head, thorax and abdomen of crippled bees, all body parts were always strongly positive for viral sequences. In contrast, for asymptomatic bees viral sequences could be detected in RNA extracted from the thorax and/or abdomen but never in RNA extracted from the head. DWV replication was demonstrated in almost all DWV-positive body parts of infected bees. Analysing individual mites for the presence of DWV revealed that the percentage of DWV-positive mites differed between mite populations. In addition, it was demonstrated that DWV was able to replicate in some but not all mites. Interestingly, virus replication in mites was correlated with wing deformity. DWV was also detected in the larval food, implicating that in addition to transmission by V. destructor DWV is also transmitted by feeding.

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