Can the examination of different types of hive samples be a non-invasive method for detection and quantification of viruses in honey bee (Apis mellifera L.) colonies?
, , , , et | 20 sept. 2023
À propos de cet article
Publié en ligne: 20 sept. 2023
Pages: 323 - 331
Reçu: 22 mars 2023
Accepté: 21 juil. 2023
DOI: https://doi.org/10.2478/jvetres-2023-0046
Mots clés
© 2023 Eliška Čukanová et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.
Fig. 1.
Viral loads of deformed wing virus (DWV) in samples of bees, pollen, hive debris and hive grid smears
| DWV | ||||
|---|---|---|---|---|
| Sample | Bees | Pollen | Debris | Smears |
| A1 1 | 1.86 × 108 | 1.03 × 107 | 4.98 × 104 | − |
| A1 2 | 5.26 × 102 | 7.60 × 105 | 2.90 × 104 | 3.50 × 105 |
| A1 3 | 2.05 × 102 | 2.49 × 106 | 1.67 × 103 | − |
| A1 4 | 1.13 × 10 | − | 2.54 × 103 | 1.01 × 105 |
| A2 1 | 1.16 × 102 | 3.57 × 105 | − | − |
| A2 2 | 9.64 × 10 | − | − | − |
| A2 3 | 4.95 × 10 | − | 9.54 × 102 | − |
| A2 4 | 7.58 × 10 | − | 7.15 × 102 | − |
| A2 5 | 1.14 ×102 | 2.67 × 105 | 2.02 × 103 | − |
| A2 6 | 8.49 × 10 | − | − | − |
| A3 1 | 7.08 × 10 | 8.58 × 105 | 5.52 × 103 | − |
| A4 1 | − | 1.42 × 106 | − | − |
| A4 2 | 2.61 × 10 | − | 3.57 × 103 | − |
| A4 3 | 1.94 × 10 | − | − | − |
| A4 4 | − | − | 6.59 × 102 | − |
| A4 5 | 3.83 × 10 | − | 1.80 × 102 | − |
| A4 6 | − | − | − | − |
| A4 7 | − | 1.11 × 106 | − | − |
| A4 8 | 4.55 × 10 | 6.59 × 106 | − | − |
| A4 9 | 7.41 × 10 | − | − | − |
| A4 10 | 2.68 × 102 | − | − | − |
| A4 11 | 2.72 × 102 | − | − | − |
| A4 12 | 1.31 × 103 | − | − | − |
| A4 13 | 6.96 × 10 | − | − | − |
| Mean | 7.75 × 106 a | 1.01 × 106 ab | 4.03E × 103 ab | 1.89 × 104 bc |
| SD | 3.80 × 107 | 2.43 × 106 | 1.14 × 104 | 7.41 ×104 |
Results of acute bee paralysis virus (ABPV) detection in bees, pollen, honey, hive debris and smears from the hive bottom board, using an end-point PCR
| ABPV | |||||
|---|---|---|---|---|---|
| Sample | Bees | Pollen | Honey | Debris | Smears |
| A1 1 | − | − | − | − | − |
| A1 2 | − | + | − | − | + |
| A1 3 | − | + | − | − | − |
| A1 4 | − | + | − | − | − |
| A2 1 | − | − | − | − | − |
| A2 2 | − | − | − | − | − |
| A2 3 | − | − | − | − | − |
| A2 4 | − | − | − | − | + |
| A2 5 | − | + | − | − | − |
| A2 6 | − | + | − | − | − |
| A3 1 | − | + | − | − | − |
| A4 1 | + | + | − | − | − |
| A4 2 | + | + | − | − | − |
| A4 3 | + | − | − | − | − |
| A4 4 | + | + | + | − | − |
| A4 5 | + | − | − | − | + |
| A4 6 | + | − | + | − | − |
| A4 7 | + | − | − | − | − |
| A4 8 | + | + | − | − | − |
| A4 9 | + | − | + | − | + |
| A4 10 | + | + | − | − | − |
| A4 11 | + | − | − | − | − |
| A4 12 | + | + | − | − | − |
| A4 13 | + | − | − | − | − |
| Positivity rate (%) | 54.2 | 50.0 | 12.5 | 0.0 | 12.5 |
Primers and probes used in the qPCR and PCR reactions
| Target virus | Method | Primer/probe | Target region | Reference |
|---|---|---|---|---|
| DWV | qPCR | qDWV-1 F,R | Helicase | 2 |
| qDWV-1 P | 21 | |||
| qDWV-2 F,R,P | RdRP | 12 | ||
| BQCV | qPCR | qBQCV F,R | 7834–8119 bp | 15 |
| qBQCV P | 20 | |||
| PCR | BQCV1 F,R | 379–700 bp | 14 | |
| SBV | qPCR | qSBV F,R,P | 5’NTR | 20 |
| ABPV | PCR | ABPV-1 F,R | RdRP | 26 |
| ABPV -2 F,R | 34–240 bp | 14 |
Viral loads of BQCV and SBV in samples of bees, pollen, honey, hive debris and hive grid smears
| BQCV | SBV | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample | Bees | Pollen | Honey | Debris | Smears | Bees | Pollen | Honey | Debris | Smears | |
| A1 1 | 4.23 × 107 | 1.58 × 107 | 2.83 × 106 | 7.21 × 105 | − | 1.76 × 104 | 5.86 × 106 | 1.01 × 106 | 8.39 × 106 | − | |
| A1 2 | 5.05 × 108 | 2.40 × 106 | 3.08 × 106 | 2.10 × 106 | − | 1.53 × 105 | 2.00 × 105 | 2.30 × 106 | 6.74 × 106 | − | |
| A1 3 | 5.24 × 107 | 3.28 × 106 | 3.22 × 106 | 1.43 × 105 | − | 1.64 × 105 | 2.06 × 106 | 3.81 × 106 | 3.98 × 106 | − | |
| A1 4 | 5.57 × 107 | 2.85 × 105 | 2.15 × 106 | 7.58 × 106 | 1.36 × 105 | 3.10 × 104 | 4.53 × 104 | 1.70 × 106 | 1.04 × 107 | − | |
| A2 1 | 5.66 × 106 | − | 1.03 × 106 | − | 7.87 × 105 | 3.95 × 104 | − | 4.96 × 105 | 4.26 × 108 | 2.27 × 104 | |
| A2 2 | 3.14 × 106 | 5.00 × 106 | 2.03 × 105 | − | − | 1.41 × 109 | 1.22 × 106 | 7.19 × 104 | 3.31 × 107 | − | |
| A2 3 | 7.56 × 106 | 5.42 × 106 | 3.78 × 105 | 5.94 × 106 | − | 9.00 × 108 | 4.41 × 106 | 2.09 × 105 | 1.08 × 109 | 1.19 × 108 | |
| A2 4 | 5.66 × 107 | 7.62 × 105 | 2.30 × 106 | − | 4.18 × 105 | 2.30 × 108 | 3.00 × 105 | 1.99 × 106 | 1.20 × 109 | 2.46 × 105 | |
| A2 5 | 2.10 × 107 | 8.70 × 106 | 2.23 × 106 | 1.27 × 106 | 7.98 × 105 | 8.48 × 109 | 1.57 × 107 | 1.79 × 106 | 2.94 × 109 | 4.41 × 106 | |
| A2 6 | 9.89 × 107 | − | 3.59 × 106 | 7.24 × 105 | 7.99 × 105 | 3.06 × 106 | 7.91 × 104 | 1.48 × 106 | 8.68 × 106 | 3.53 × 105 | |
| A3 1 | 1.40 × 107 | 2.81 × 106 | 4.17 × 106 | 7.70 × 106 | − | 4.73 × 105 | 1.08 × 106 | 2.33 × 106 | 5.06 × 107 | 2.14 × 104 | |
| A4 1 | 3.94 × 107 | 3.71 × 107 | 4.94 × 106 | − | − | 2.96 × 105 | 8.30 × 106 | 8.07 × 105 | − | − | |
| A4 2 | 1.90 × 108 | 2.37 × 107 | 1.75 × 106 | 2.63 × 106 | 2.28 × 105 | 4.20 × 108 | 4.20 × 106 | 1.71 × 105 | 8.60 × 107 | − | |
| A4 3 | 6.86 × 107 | 2.62 × 107 | 1.69 × 105 | − | − | 1.01 × 105 | 4.21 × 106 | 4.26 × 104 | 1.40 × 107 | − | |
| A4 4 | 4.74 × 107 | 5.59 × 106 | 3.40 × 105 | − | − | 2.09 × 105 | 1.33 × 106 | 6.47 × 104 | 1.64 × 107 | − | |
| A4 5 | 1.16 × 108 | 1.52 × 107 | 3.57 × 106 | 1.05 × 106 | − | 1.22 × 105 | 3.29 × 106 | 6.65 × 105 | 1.08 × 106 | 8.41 × 103 | |
| A4 6 | 4.93 × 107 | − | 4.86 × 106 | − | 6.87 × 104 | 2.91 × 105 | − | 1.69 × 106 | 5.34 × 106 | − | |
| A4 7 | 1.07 × 108 | 3.50 × 107 | 1.48 × 106 | − | − | 1.16 × 105 | 4.60 × 106 | 2.38 × 105 | − | − | |
| A4 8 | 1.48 × 108 | 8.35 × 107 | 1.10 × 106 | − | 5.03 × 105 | 1.12 × 105 | 4.61 × 106 | 1.07 × 105 | − | 1.05 × 105 | |
| A4 9 | 1.62 × 108 | 2.45 × 107 | 7.64 × 104 | − | 2.42 × 105 | 6.43 × 108 | 1.95 × 106 | 1.58 × 104 | − | − | |
| A4 10 | 9.39 × 107 | 2.40 × 107 | 1.90 × 106 | − | − | 3.51 × 105 | 2.93 × 106 | 7.64 × 105 | − | − | |
| A4 11 | 7.00 × 107 | 2.25 × 107 | 1.16 × 107 | − | − | 4.20 × 105 | 2.19 × 107 | 4.43 × 106 | − | 9.20 × 103 | |
| A4 12 | 2.82 × 108 | 3.74 × 107 | 3.55 × 105 | − | 1.60 × 105 | 4.10 × 105 | 1.10 × 107 | 5.64 × 104 | − | 4.56 × 104 | |
| A4 13 | 1.00 × 108 | 3.32 × 107 | 1.15 × 106 | 5.84 × 105 | 2.59 × 105 | 1.72 × 109 | 8.10 × 106 | 2.88 × 105 | 4.19 × 108 | 7.95 × 105 | |
| Mean | 9.73 × 107 a | 1.72 × 107 b | 2.44 × 106 bc | 1.27 × 106 cd | 1.83 × 105 d | 5.75 × 108 a | 4.47 × 106 a | 1.11 × 106 a | 2.63 × 108 a | 5.21 × 106 b | |
| SD | 1.09 × 108 | 1.93 × 107 | 2.46 × 106 | 2.37 × 106 | 2.75 × 105 | 1.75 × 109 | 5.38 × 106 | 1.22 × 106 | 6.58 × 108 | 2.43 × 107 | |
Results of virus detection in bees, pollen, honey, hive debris and hive grid smears, using an end-point PCR (black queen cell virus (BQCV)) and a qPCR (deformed wing virus (DWV), black queen cell virus (BQCV) and sacbrood virus (SBV))
| DWV | BQCV | SBV | |||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample | Bees | Pollen | Honey | Debris | Smears | Bees | Pollen | Honey | Debris | Smears | Bees | Pollen | Honey | Debris | Smears |
| A1 1 | + | + | + | + | - | + | + | + | + | + | + | + | + | + | − |
| A1 2 | + | + | + | + | + | + | + | + | + | − | + | + | + | + | − |
| A1 3 | + | + | + | + | − | + | + | + | + | − | + | + | + | + | − |
| A1 4 | + | − | + | + | + | + | + | + | + | + | + | + | + | + | − |
| A2 1 | + | + | − | − | − | + | − | + | − | + | + | − | + | + | + |
| A2 2 | + | − | − | − | − | + | + | + | − | − | + | + | + | + | − |
| A2 3 | + | + | − | + | − | + | + | + | + | − | + | + | + | + | + |
| A2 4 | + | − | − | − | − | + | + | + | − | + | + | + | + | + | + |
| A2 5 | + | + | − | + | − | + | + | + | + | + | + | + | + | + | + |
| A2 6 | + | − | + | + | − | + | − | + | + | + | + | + | + | + | + |
| A3 1 | + | + | + | + | − | + | + | + | + | + | + | + | + | + | + |
| A4 1 | + | + | + | − | − | + | + | + | − | − | + | + | + | − | − |
| A4 2 | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
| A4 3 | + | + | − | − | − | + | + | + | − | − | + | + | + | + | − |
| A4 4 | + | + | − | + | − | + | + | + | − | − | + | + | + | + | − |
| A4 5 | + | + | + | + | + | + | + | + | + | − | + | + | + | + | + |
| A4 6 | + | − | + | − | − | + | − | + | − | + | + | − | + | + | − |
| A4 7 | + | + | + | − | − | + | + | + | − | − | + | + | + | − | − |
| A4 8 | + | + | − | − | − | + | + | + | − | + | + | + | + | − | + |
| A4 9 | + | + | − | − | − | + | + | + | − | + | + | + | + | − | − |
| A4 10 | + | − | + | − | − | + | + | + | − | − | + | + | + | − | − |
| A4 11 | + | + | + | − | − | + | + | + | − | − | + | + | + | − | + |
| A4 12 | + | + | + | − | − | + | + | + | − | + | + | + | + | − | + |
| A4 13 | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
| Positivity rate (%) | 100.0 | 75.0 | 62.5 | 50.0 | 20.8 | 100.0 | 87.5 | 100.0 | 45.8 | 54.2 | 100.0 | 91.7 | 100.0 | 70.8 | 50.0 |