natgillin/translations-raw huggingface dataset:
The sections shall be counter-stained with Bismarck Brown Yellow, rinsed in dH2O, and cover-slips shall be applied using an aqueous mounting medium.
Prečiščena DNK se zamrzne pri – 20 °C, če se ne uporabi takoj.
Positive and negative controls shall be sections from known infected and non-infected hosts respectively.
Sekvenciranje produktov PCR
A positive result shall be demonstrated by the purple-black labelling of M. refringens cells within known target tissues, with all negative controls being negative and all positive controls being positive.
DNK se sekvencira s kompletom Big Dye Terminator Kit v3,1 (Applied Biosystems).
Sequencing
Skupna količina pri vsaki reakciji je 20 μl, končne koncentracije pa so 1 × Big Dye Terminator, 1 × pufer za sekvenciranje, 10 pmol/μl smiselnega ali protismiselnega začetnega oligonukleotida in 10 μl prečiščene DNK (razredčeno na približno 10 ng/μl), kar se analizira pod naslednjimi pogoji pomnoževanja na cikličnem termostatu DNA Engine Tetrad 2 Peltier Thermal Cycler (ali enakovrednem termostatu): 30 sekund pri 94 °C, nato 10 sekund pri 96 °C, 10 sekund pri 50 °C in štiri minute pri 60 °C, pri čemer se zadnji trije koraki 30-krat ponovijo.
Sequencing shall be carried out as one of the final steps for confirmatory diagnostic.
Produkti PCR se oborijo z metodo z natrijevim acetatom, pri kateri se 10 μl NaAc, 70 μl H2O in 250 μl etanola doda 20 μl DNK, premeša v vorteks mešalniku in 20 minut centrifugira pri 13000 vrtljajih na minuto, supernatant se odstrani, peleti se sperejo z 200 μL absolutnega etanola, nato pa sledi petminutno centrifugiranje pri 13000 vrtljajih na minuto.
Targeted regions are the SSU rDNA and ITS1.
Peleti se pet minut sušijo pri 37 °C.
Detailed diagnostic methods and procedures for surveillance and confirmation of infection with Marteilia refringens
Doda se jim 25 μl Hi-Di formamida, dve minuti se segrevajo na 95 °C in temeljito premešajo v vorteks mešalniku.
For the purpose of surveillance programmes and to confirm the presence of an infection with Marteilia refringens or to rule out a suspicion of that listed disease, in accordance with the requirements laid down in Section II of Part 4 to Annex I, the diagnostic methods and corresponding procedures to be used shall be in accordance with the guidelines laid down in Table 4.1 as follows:
Vzorci se sekvencirajo z analizatorjem ABI3130xl Avant Genetic Analyser v skladu z navodili proizvajalca.
Table 4.1
Rezultati sekvenciranja se analizirajo s programsko opremo Sequencher, sekvence pa se primerjajo s sekvencami iz podatkovne baze NCBI s funkcijo BLAST.
Guidelines for the use of diagnostic methods for the surveillance programmes and to confirm or rule out infection with Marteilia refringens
NADZORNE IN KONTROLNE METODE ZA VIRUSNO HEMORAGIČNO SEPTIKEMIJO (VHS) IN INFEKCIOZNO HEMATOPOETSKO NEKROZO (IHN)
Method
I.2 Posebne zahteve za pridobitev zdravstvenega statusa brez bolezni (kategorija I) glede VHS in IHN
Targeted surveillance
I.2.1 Programi nadzora:
Presumptive diagnosis
I.2.2 Programi izkoreninjenja
Confirmatory diagnosis
I.2.2.1 Splošne zahteve
Digestive gland imprints
I.2.2.2 Zahteve za ponovno pridobitev statusa brez bolezni za celinske kompartmente, ki zajemajo eno samo gojilnico, predhodno razglašeno za prosto IHN ali VHS ali obeh bolezni
X, or
I.3 Posebne zahteve za ohranitev zdravstvenega statusa brez bolezni (kategorija I) glede VHS ali IHN ali obeh bolezni
Histopathology
I.4 Zahteve za odpravo ukrepov obvladovanja iz člena 39 Direktive 2006/88/ES, in sicer za spremembo zdravstvenega statusa iz kategorije V v kategorijo III
In situ hybridisation
v gojilnici, za katero obstaja sum okužbe, se opravita vsaj en zdravstveni pregled in eno vzorčenje 10 rib, ko so opaženi klinični znaki ali post mortem znaki, ki ustrezajo znakom okužbe z VHS ali IHN ali obema boleznima, ali vsaj 30 rib, ko klinični znaki ali post mortem znaki niso opaženi.
X, and
Vzorci se testirajo z uporabo ene ali več diagnostičnih metod iz točk (i) in (ii) v skladu z natančnimi diagnostičnimi metodami in postopki, kot so določeni v oddelku II dela 1 Priloge II:
DETAILED DIAGNOSTIC METHODS AND PROCEDURES FOR THE SURVEILLANCE AND CONFIRMATION OF INFECTION WITH BONAMIA OSTREAE
I.2 Posebne zahteve za dosego zdravstvenega statusa brez bolezni (kategorija I) glede KHVD
I. Procedures for diagnosis of Infection with Bonamia ostreae
I.2.1 Programi nadzora
When sampling and laboratory examination are carried out for the purpose of obtaining or maintaining a certain health status with regard to Bonamia ostreae as set out in Section I of Part 5 of Annex I or to confirm or rule out the presence of that listed disease in accordance with Article 57(b) of Directive 2006/88/EC, using the diagnostic methods set out in Section II of Part 5 of Annex I, the detailed diagnostic methods and procedures set out in following points I.1, I.2 and I.3 shall apply.
I.2.2.2 Zahteve za ponovno pridobitev statusa brez bolezni za celinske kompartmente, ki zajemajo eno samo gojilnico, predhodno razglašeno za prosto KHVD
Sample process
I.3 Posebne zahteve za ohranitev statusa kategorije I glede KHVD
Once sampled, oysters shall be maintained at 4 °C or on refrigerated ice for no more than 24 hours if samples include gaping molluscs and 72 hours if not, in a plastic bag including a label with details related to the nature and the origin of the oysters.
I.4 Posebne zahteve za odpravo ukrepov obvladovanja iz člena 39 Direktive 2006/88/ES za pridobitev zdravstvenega statusa kategorije III glede KHVD v državah članicah, na območjih ali v kompartmentih z zdravstvenim statusom kategorije V
A 3 to 5 mm thick section of tissues including gills and heart tissue shall be used for the diagnostic of Bonamia ostreae by histology.
I.2 Posebne zahteve za dosego zdravstvenega statusa kategorije I glede ISA
Slides shall be air-dried, fixed in methanol or in absolute ethanol and stained using a commercially available blood-staining kit (namely such as Diff- Quik®/Hemacolor®.), in accordance with the manufacturer's instructions.
I.3 Minimalni kontrolni ukrepi za ohranitev statusa kategorije I glede ISA
Slides shall be observed first at × 200 magnification and then under oil immersion at × 1000 magnification.
I.4 Posebne zahteve za zdravstveni status kategorije III glede ISAV z delecijo HPR v državah članicah, na območjih ali v kompartmentih s predhodnim zdravstvenim statusom kategorije V
A positive result shall be the presence of small spherical or ovoid organisms (2 to 5 μm wide) within haemocytes.
če so v proizvodnih enotah ali na proizvodnem območju školjke rodu Ostrea spp. in Mytilus spp., se vzorčijo enako veliki vzorci obeh rodov.
However, the parasite might also occur extracellularly.
Če je prisoten le eden od obeh rodov, se vzorči navedeni rod.
These organisms show a basophilic cytoplasm and an eosinophilic nucleus (colours may vary with the stain used) and, because they spread on the slide, they can appear wider on imprints than on histological examination.
Če ni prisoten niti rod Ostrea niti rod Mytilus, mora biti vzorec reprezentativen za vse druge prisotne dovzetne vrste;
Multinucleated cells may be observed.
če so v proizvodnih enotah oslabeli, odprti ali pred kratkim poginuli, toda ne razkrojeni mehkužci, se primarno izberejo takšni mehkužci.
Sections of tissue that include gills and digestive gland, shall be fixed for a period of at least 24 hours in Davidson's fixative followed by normal processing for paraffin histology and staining, for example with haematoxylin and eosin.
Če takšnih mehkužcev ni med njimi, izbrani mehkužci vključujejo najstarejše zdrave mehkužce;
A positive result shall be the presence of parasites as very small cells of 2 to 5 μm wide within the haemocytes or free in the connective tissue or sinuses of gill, gut and mantle epithelium, often associated with an intense inflammatory reaction.
pri vzorčenju v gojilnicah mehkužcev, v katerih se za gojenje mehkužcev uporablja več kot en vodni vir, se mehkužci iz vseh vodnih virov vključijo v vzorčenje tako, da so v vzorcu sorazmerno zastopani vsi deli gojilnice;
To avoid any doubt, the parasite shall be observed inside the haemocyte for a positive diagnosis.
pri vzorčenju na območjih gojenja mehkužcev se mehkužci iz zadostnega števila mest vzorčenj vključijo v vzorčenje tako, da so v vzorcu sorazmerno zastopani vsi deli območja gojenja mehkužcev.
DNA extraction kits, which are available commercially and usually produce high quality DNA suitable for use with the PCR protocols as detailed below, may be used.
Glavni dejavniki, ki jih je treba upoštevati pri izbiri teh mest vzorčenja, so mesta vzorčenja s predhodnim dokazom Marteilia refringens, gostota živali, vodni tokovi, prisotnost dovzetnih vrst, prisotnost vektorskih vrst, batimetrija in prakse upravljanja.
Two PCR protocols targeting the small subunit (SSU) rDNA may be used:
V vzorčenje se vključijo naravna nahajališča.
the first one is a conventional PCR that amplify several members of the Haplosporidia including Bonamia spp.
I.2 Posebne zahteve za dosego zdravstvenega statusa kategorije I glede Marteilia refringens
Primers designated Bo and Boas are 5′-CAT-TTA-ATT-GGT-CGG-GCC-GC-3′ and 5′-CTG-ATC-GTC-TTC-GAT-CCC-CC-3′ respectively, and amplify a 300 bp product.
I.3 Posebne zahteve za ohranitev zdravstvenega statusa brez bolezni (kategorija I) glede okužbe z Marteilia refringens
PCR mixtures contain buffer (500 mM KCl, 100 mM Tris/HCl [pH 9,0 at 25 °C] and 1 % Triton® X-100), 2,5 mM MgCl2, 0,2 mM dNTP mix, 1 μM forward and reverse primers, 0,02 units μl– 1 Taq DNA polymerase, and 0,2 ng μl– 1 of the DNA template in a total volume of 50 μl.
I.4 Zahteve za odpravo ukrepov obvladovanja, določenih v členu 39 Direktive 2006/88/ES (sprememba zdravstvenega statusa iz kategorije V v kategorijo III) glede okužbe z Marteilia refringens
Samples shall be denatured in a thermocycler for five minutes at 94 °C before being submitted to 30 cycles (94 °C for one minute, 55 °C for one minute, 72 °C for one minute) followed by a final extension of 10 minutes at 72 °C.
če so med njimi ostrige vrste Ostrea edulis, se za vzorčenje izberejo samo ostrige navedene vrste.
A positive result shall be positive PCR amplification at the expected size (namely, 300 bp), with all negative controls being negative and all positive controls being positive;
Če med njimi ni ostrig vrste Ostrea edulis, je vzorec reprezentativen za vse druge prisotne dovzetne vrste;
the second PCR protocol is a SYBR® Green Real time PCR assay.
če so med njimi oslabeli, odprti ali pred kratkim poginuli, toda ne razkrojeni mehkužci, se primarno izberejo takšni mehkužci.
It allows the specific detection of B. ostreae (described below) and may be combined with a SYBR® Green Real time PCR assay allowing the specific detection of B. exitiosa (Ramilo et al.
pri vzorčenju v gojilnicah, kjer se za gojenje mehkužcev uporablja več kot en vodni vir, se mehkužci iz vseh vodnih virov vključijo v vzorčenje tako, da so v vzorcu sorazmerno zastopani vsi deli gojilnice;
2013).
pri vzorčenju na območjih gojenja mehkužcev se v vzorec vključijo mehkužci iz zadostnega števila mest vzorčenja.
Primers BOSTRE-F (5′- TTACGTCCCTGCCCTTTGTA-3′) and BOSTRE-R (5′- TCGCGGTTGAATTTTATCGT-3′) amplify a 208 bp product.
Glavni dejavniki, ki jih je treba upoštevati pri izbiri navedenih mest vzorčenja, so mesta vzorčenja s predhodnim dokazom Bonamia ostreae, gostota živali, vodni tokovi, prisotnost dovzetnih vrst, prisotnost vektorskih vrst, batimetrija in prakse upravljanja.
PCR mixtures contain SYBR® Green Master Mix (1X), 0,3 μM forward and reverse primers, and 200 ng of extracted DNA.
V vzorčenje se vključijo naravna nahajališča, ki se nahajajo znotraj območja gojenja ali ob njem.
Samples shall be denatured in a Real Time Detection System for 10 minutes at 95 °C before being submitted to 35 cycles (95 °C for 30 seconds, 55 °C for 45 seconds and 72 °C for one minute).
I.2 Posebne zahteve za dosego zdravstvenega statusa kategorije I glede Bonamia ostreae
Melting temperature curve analysis shall be carried out with temperature increments of 0,5 °C/s starting at 55 °C and ending at 95 °C and recording fluorescence at each change of temperature.
I.3 Posebne zahteve za ohranitev zdravstvenega statusa brez bolezni (kategorija I) glede okužbe z Bonamia ostreae
Positive controls shall consist in genomic DNA from a highly infected host or plasmidic DNA including the target region.
I.4 Zahteve za odpravo ukrepov obvladovanja, določenih v členu 39 Direktive 2006/88/ES (sprememba zdravstvenega statusa iz kategorije V v kategorijo III) glede okužbe z Bonamia ostreae
Negative controls shall consist in genomic DNA from non-infected hosts and PCR reactives without target DNA.
I.2 Posebne zahteve za pridobitev zdravstvenega statusa kategorije I glede WSD
A positive result shall be positive PCR amplification with a unique melting temperature peak (78,25 ± 0,25 °C in the conditions published by Ramilo et al.
I.2.2.2 Zahteve za ponovno pridobitev statusa brez bolezni glede WSD za celinske kompartmente, ki zajemajo eno samo gojilnico, predhodno razglašeno za prosto WSD
2013), with all negative controls being negative and all positive controls being positive.
I.4 Zahteve za odpravo ukrepov obvladovanja, določenih v členu 39 Direktive 2006/88/ES (sprememba zdravstvenega statusa iz kategorije V v kategorijo III) glede WSD
A probe that targets the SSU of the rDNA gene complex shall be used although it was shown to cross react with some other members of the family Haplosporidia.
I.1 Priprava in pošiljanje vzorcev rib
Sections of tissue that include gills and digestive gland shall be fixed for a period of at least 24 hours in Davidson's fixative followed by normal processing for paraffin histology.
I.1.1 Tkiva za virološko preiskavo na celični kulturi
Sections shall be incubated with 100 μl of hybridisation buffer (4 × SSC [standard saline citrate], 50 % formamide, 1 × Denhardt's solution, 250 μg ml– 1 yeast tRNA, 10 % dextran sulphate) containing 20 ng (2 μl of the PCR reaction prepared as described in point I.3.2 using primers Bo and Boas) of the digoxigenin-labelled probe.
I.2 Pošiljanje vzorcev za ribe
Sections shall be washed twice for five minutes in 2 × SSC at room temperature, and once for 10 minutes in 0,4 × SSC at 42 °C.
I.3 Zbiranje dodatnega diagnostičnega materiala
A positive result shall correspond to labelled parasites inside the haemocytes, with all negative controls negative and all positive controls positive.
I.4 Priprava vzorcev za preiskavo celične kulture in RT-qPCR
Targeted regions shall be the SSU rDNA and ITS1.
I.4.1 Zamrznitev v izjemnih primerih
Procedures for surveillance and confirmation of infection with Bonamia ostreae
I.4.2 Homogenizacija organov
For the purpose of surveillance and confirmation of the presence of or to rule out a suspicion of infection with Bonamia ostreae in accordance with the requirements laid down in Section II of Part 5 to Annex I, the diagnostic methods and corresponding procedures to be used shall be in accordance with the guidelines laid down in the following Table 5.1.
I.4.3 Centrifugiranje homogenata
Table 5.1
I.4.4 Priprava vzorca za programe nadzora, ki temeljijo na RT-PCR in RT-qPCR
Guidelines for the use of diagnostic methods for the surveillance programmes and to rule out or confirm infection with Bonamia ostreae
I.4.5 Združevanje vzorcev za RT-qPCR
Heart or gill imprints
I.5 Virološka preiskava na celični kulturi
DETAILED DIAGNOSTIC METHODS AND PROCEDURES FOR SURVEILLANCE AND CONFIRMATION OF WHITE SPOT DISEASE (WSD)
I.5.1 Celične kulture in gojišča
Diagnostic procedures for detection of WSSV
Gojišče, ki se uporablja za gojenje celic v odprtih enotah, se lahko puferira s tris(hidroksimetil)aminometan-HCl (Tris-HCl) (23 mM) in natrijevim bikarbonatom (6 mM).
When sampling and laboratory examination for the purpose of the surveillance and eradication programmes set out in Section I of Part 6 of Annex I and for the confirmation of the presence of or to rule out a suspicion of infection with WSSV are carried out in accordance with Article 57(b) of Directive 2006/88/EC, using the diagnostic methods set out in Section II of Part 6 of Annex I, the detailed diagnostic methods and procedures set out in points 2 to 7 of this Part shall apply.
Vrednost pH mora biti 7,6 ± 0,2.
The methods and procedures described in this Part of Annex II are adapted from the ISO 17025 accredited test applied at the European Union Reference Laboratory for Crustacean Diseases.
I.5.2 Inokulacija celičnih kultur
Alternative approaches, using equivalent conditions or kits produced by different manufacturers, but which offer equivalent sensitivity and specificity to those described in this Part may be applied.
I.5.3 Inkubacija celičnih kultur
In all instances, PCR amplified product shall be sequenced to confirm identity as White spot syndrome virus (WSSV).
I.5.4 Mikroskopija
Tissue (pleopods and gills) containing WSSV from crustaceans may be stored in ethanol, RNAlater or snap frozen at – 80 °C.
I.5.5 Subkultivacija
The stages required for the identification of WSSV from tissue samples shall be as follows: Homogenisation of the tissue, extraction of the DNA, specific amplification of WSSV DNA using PCR, visualisation of the amplified product on a gel, purification of the DNA and sequencing to confirm the identity of the pathogen.
I.6.1 ELISA
Tissue homogenisation
I.6.2 Imunofluorescenca – IF
The disruption of the tissues and the preparation of a homogenate in an appropriate buffer shall be carried out using the Fast Prep tissue disruptor and Lysing matrix A tubes (MP Biomedicals).
I.6.3 Nevtralizacija
The tissue shall be weighed, placed in Lysing Matrix A tubes, diluted 1 in 10 w/v or according to manufacturer's instructions, in an appropriate buffer (G2 and 10 μl Proteinase K for use with DNA Tissue kit (Qiagen)) and homogenised using the Fast Prep 24 homogeniser for two minutes.
I.6.4 RT-PCR/ RT-qPCR
Homogenised samples shall be incubated at 56 °C for a minimum period of four hours or overnight.
I.6.4.1 Priprava virusne RNK
Samples shall be vortexed, centrifuged at 9000 rpm for two minutes and 50 μl of supernatant or a volume equivalent to 5 mg of tissue (weight of tissue optimal for extraction kit) shall be added to a sample tube for DNA extraction and volume made up to 200 μl using G2 buffer.
I.6.4.3 RT-qPCR za VHSV
Total DNA shall be extracted using a DNA tissue extraction kit and the EZ1 Advanced XL Biorobot (Qiagen) following the manufacturers' instructions.
I.6.4.4 RT-qPCR za IHNV
An extraction control (Calf Thymus DNA) and a negative control (G2 buffer) shall run with each batch of samples.
RT-qPCR
DNA shall be eluted into 50 μl volume.
I.1 Priprava vzorcev iz rib
To ensure that the extraction has completed successfully, the DNA concentration for all samples and controls shall be determined using a Nano Drop machine.
I.1.1 Ekstrakcija DNK
Extracted DNA shall be frozen at – 20 °C if not required immediately.
I.2 Dokazovanje in določanje povzročitelja z metodo na podlagi verižne reakcije s polimerazo (PCR)
WSSV Polymerase Chain Reaction (PCR)
I.2.2 Konvencionalni test PCR za dokazovanje KHV
The method to be used for the detection of WSSV shall be the protocol for the detection of WSSV by nested PCR set out in the following paragraphs which amplifies a 1447 bp and an 848 bp amplicon of the 18s rRNA gene in the first and second round PCRs respectively.
I.2 Pošiljanje vzorcev iz rib
The 1st round PCR reaction shall be set up in a 50 μl volume which contains final concentrations of 1 × GoTaq Buffer (Promega), 5 mM MgCl2, 1 pmol/μl WSSV 146 F1 primer, 1 pmol/μl WSSV 146 R1 primer (Table 1), 0,25 mM dNTPs, 1,25U Taq polymerase and 2,5 μl of DNA.
I.1 Postopek vzorčenja
Each sample shall be run in duplicate alongside a negative extraction control, a negative PCR control, (2,5 μl H2O added instead of DNA) and a positive control.
I.2 Mikroskopske metode
The positive control shall be diluted WSSV plasmid made and validated for use in-house (available from the EURL).
I.2.1 Citologija (citologija odtisa)
The 2nd round WSSV PCR reaction shall be set up in the same way as the first round but using the WSSV 146 F2/R2 primer set and using a second positive control to check that this stage of the PCR has worked.
I.2.2 Histologija
Primer
I.3 Molekularne metode
Both 1st and 2nd round PCRs shall be run using the following cycling conditions on a DNA Engine Tetrad 2 Peltier Thermal Cycler (or equivalent).
I.3.1 Ekstrakcija DNK
An initial denaturation step of 94 °C for two minutes followed by 94 °C for 30 seconds, 62 °C for 30 seconds, 72 °C for 30 seconds repeated for 30 cycles, a 72 °C extension step for two minutes and held at 4 °C.
I.3.2 Verižna reakcija s polimerazo (PCR)
Gel electrophoresis
I.3.3 Hibridizacija in situ (ISH)
Amplified PCR products from both the 1st and 2nd round PCRs shall be visualised on 2 % agarose gels made using TAE buffer.
I.3.4 Sekvenciranje
Samples of that size shall be cut out and placed into a 1,5 ml microfuge tube.
Začetna oligonukleotida Bo in Boas sta 5′-CAT-TTA-ATT-GGT-CGG-GCC-GC-3′ oz. 5′-CTG-ATC-GTC-TTC-GAT-CCC-CC-3′ in amplificirata produkt 300 bp.
The DNA contained within the gel slices shall be purified using Promega Wizard® SV Gel and PCR clean-Up System according to manufacturers' instructions.
Če titer katerega koli od treh izolatov virusa v primerjavi z začetnim titrom pade za faktor log 2 ali več, se celična linija ne uporablja več za namene nadzora.
The concentration of the DNA shall be estimated using a Nano Drop machine.
Če se v laboratoriju hranijo različne celične linije, se vsaka linija pregleda ločeno.
The purified DNA shall be frozen at – 20 °C if not being used immediately.
Evidenca se hrani vsaj 10 let.
Sequencing of PCR products
I. Natančne diagnostične metode in postopki za potrditev prisotnosti ali izključitev suma na KHVD
DNA shall be sequenced using the Big Dye Terminator Kit v3,1 (Applied Biosystems).
I. Zahteve za programe nadzora in izkoreninjenja za pridobitev in ohranitev zdravstvenega statusa brez bolezni glede okužbe z Marteilia refringens
The total volume in each reaction is 20 μl, the final concentrations being 1 × Big Dye terminator, 1 × sequencing buffer, 10 pmol/μl forward or reverse primer and 10 μl of purified DNA (diluted to approximately 10 ng/μl) run using the following cycling conditions on a DNA Engine Tetrad 2 Peltier Thermal Cycler (or equivalent): 94 °C for 30 seconds followed by 96 °C for 10 seconds, 50 °C for 10 seconds and 60 °C for four minutes, cycling the last three steps 30 times.
I.1.2 Vzorci za verižno reakcijo s polimerazo z reverzno transkripcijo (RT-PCR ali RT-qPCR)
The PCR products shall be precipitated using a sodium acetate method where 20 μl DNA is added to 10 μl NaAc, 70 μl H2O and 250 μl ethanol, vortexed and centrifuged at 13000 rpm for 20 minutes, the supernatant shall be removed and the pellet shall be washed with 200 μl absolute ethanol, centrifuging at 13000 rpm for five minutes.
I.6 Določanje virusa
The pellet shall be dried for five minutes at 37 °C.
vse gojilnice, uradno razglašene za okužene z ISAV z delecijo HPR ali s potrjeno boleznijo ISA, se izpraznijo, očistijo in razkužijo, njihova proizvodnja pa se prekine za vsaj tri mesece. okužena in ogrožena območja se lahko prekličejo, ko se vse gojilnice znotraj okuženega območja izpraznijo, očistijo in razkužijo, sledi pa jim sinhronizirana prekinitev proizvodnje vsaj šest tednov.
Samples shall be sequenced using the ABI3130xl Avant Genetic analyser according to manufacturers' instructions.
I. Zahteve za programe nadzora in izkoreninjenja za pridobitev in ohranitev zdravstvenega statusa brez bolezni glede KHVD ter za obvladovanje okužbe s koi herpes virusom (KHV)
Sequencing results shall be analysed using Sequencher software and sequences matched to sequences on the NCBI database using the BLAST function.
I. Zahteve za programe nadzora in izkoreninjenja za pridobitev in ohranitev zdravstvenega statusa brez bolezni glede ISA ter za obvladovanje okužbe z ISAV z delecijo HPR
SURVEILLANCE AND CONTROL METHODS FOR VIRAL HAEMORRHAGIC SEPTICAEMIA (VHS) AND INFECTIOUS HEMATOPOIETIC NECROSIS (IHN)
I. Zahteve za programe nadzora ali izkoreninjenja za pridobitev in ohranitev zdravstvenega statusa brez bolezni glede okužbe z Bonamia ostreae