Fig. 1.
Fig. 2.
Fig. 3.
Assessments of percentage sequence similarities according to GenBank for PCR products of amplification of a disease aetiological agent isolated from different fishes
| Fish | Acipenser ruthenus | AciHV-3 | Remarks |
|---|---|---|---|
| Common carp | 96% | 99% | |
| Tench | 97% | 99% | |
| Crucian carp 1 | 97% | 99% | AciHV-3-like |
| Crucian carp 2 | 95% | 96% | |
| Sturgeon 1 | 97% | 99% | |
| Sturgeon 2 | 90% | 92% | Suspicious |
| Sturgeon 3 | 96% | 98% | |
| Sturgeon 4 | 97% | 99% | |
| Sturgeon 5 | 97% | 99% | AciHV-3-like |
| Sturgeon 6 | 96% | 98% | |
| Sturgeon 7 | 97% | 99% | |
| Sturgeon 8 | 97% | 99% | |
| Average value | 96% | 98% | |
| Median value | 97% | 99% |
Primer pairs used in this study for DNA and RNA extraction from Russian sturgeon in a mortality event and from cohabited tench, Nile tilapia, crucian carp and common carp
| Name | Sequence | Reference |
|---|---|---|
| Stör-HV-1F | 5′-CGG AAT TCT AGA YTT YGC IWS IYT ITA YCC-3′ | (9) |
| Stör-HV-1R | 5′-CCC GAA TTC AGA TCT CIG TRT CIC CRT A-3′ | |
| Stör-HV-Fn | 5′-GCA GCA GAC TAC GTG GTG TAC G-3′ | (9) |
| Stör-HV-Rn | 5′-AGT TGG RGC AGA TCT TCA TTT CGG-3′ | |
| Terminase-AciHV-1 F | 5′-ACC TCG TGT TGA TCG-3′ | (15) |
| Terminase-AciHV-1 R | 5′-TCA AAA CTT CCG GGT-3′ | |
| Term F2 | 5′-GCM MGR GGA CAG AWC CCM G-3′ | (15) |
| Termsal-3 Rdeg | 5′-GGT GCA CAC RCC MAD IGA CG-3′ | |
| TermExon 1F | 5′-CAG GTB GAR CTM ATG MGR GGG TTT TT-3′ | personal |
| TermExon 1R | 5′-CAT RAT KKY DGT YTT VCC RCA YTG TC-3′ | communication |
Preparation of master mixes for first-round PCRs using a 5 μL template to amplify DNA of an aetiological agent infecting Russian sturgeon lethally
| Components | volume × 1 (in μL) |
|---|---|
| Diethyl pyrocarbonate–treated water | 11.7 |
| 5× Green GoTaq Flexi Puffer (Promega) | 5 |
| MgCl2 25 mM (Promega) | 2.5 |
| dNTPs 10 mM (Thermo Fisher) | 0.25 |
| GoTaq DNA polymerase (Promega) | 0.125 |
| Primer premix (1:1) 400 nmol | 0.5 |
RNA and DNA extraction kit concentration of DNA from an aetiological agent infecting Russian sturgeon lethally
| Sturgeon sample numbers | RNA extraction kit double-stranded DNA in ng/μL* | DNA extraction kit |
|---|---|---|
| 388 | 554.90 | 893.05 |
| 544 | 150.65 | 1220.65 |
| 577 | 889.55 | 889.05 |
| 608 | 240.95 | 2051.75 |
| Average | 459.0125 | 1263.625 |
Positive sturgeon herpesvirus PCRs in different fish samples
| Species | Positive cases/total | Rate (%) | Remarks |
|---|---|---|---|
| Sturgeon | 105/120 | 87.5 | Infected |
| Tench | 2/5 | 40 | Infected |
| Crucian carp | 4/6 | 66.6 | Infected |
| Common carp | 2/6 | 33.3 | Suspicious but positive |
| Tilapia | 0/4 | - | Negative |
Investigation of sturgeon organ tropism using PCR for sturgeon herpesvirus
| Organ | Sampling | ||||
|---|---|---|---|---|---|
| 1 (n = 359) | 2 (n = 360) | 3 (n = 361) | 4 (n = 362) | 5 (n = 363) | |
| Heart | + | + | + | - | + |
| Liver | + | + | + | - | + |
| Spleen | - | + | + | - | + |
| Kidney | - | - | + | - | + |
| Pancreas | - | + | + | - | + |
| Swim bladder | - | + | + | - | + |