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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? Cover

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?

Journal of Veterinary Research
Volume 67 (2023): Issue 3 (September 2023)
By: ,  ,  ,  ,   and    
Open Access
|Sep 2023
Figures & tables

Figures & Tables

Fig. 1.

Map indicating the sample collection area within the Czech Republic (a) and locations of the four sample collection sites (Apiaries 1–4) along with their respective GPS locations (b)

Table 1.

Primers and probes used in the qPCR and PCR reactions

Target virusMethodPrimer/probeTarget regionReference
DWVqPCRqDWV-1 F,RHelicase2
qDWV-1 P21
qDWV-2 F,R,PRdRP12
BQCVqPCRqBQCV F,R7834–8119 bp15
qBQCV P20
PCRBQCV1 F,R379–700 bp14
SBVqPCRqSBV F,R,P5’NTR20
ABPVPCRABPV-1 F,RRdRP26
ABPV -2 F,R34–240 bp14

1 bp – base pairs; DWV – deformed wing virus; BQCV – black queen cell virus; SBV – sacbrood virus; ABPV – acute bee paralysis virus; F – forward; R – reverse; P – probe

Table 2.

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))

DWVBQCVSBV
SampleBeesPollenHoneyDebrisSmearsBeesPollenHoneyDebrisSmearsBeesPollenHoneyDebrisSmears
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 (%)100.075.062.550.020.8100.087.5100.045.854.2100.091.7100.070.850.0
Table 3.

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
SampleBeesPollenHoneyDebrisSmears
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 (%)54.250.012.50.012.5
Table 4.

Viral loads of deformed wing virus (DWV) in samples of bees, pollen, hive debris and hive grid smears

DWV
SampleBeesPollenDebrisSmears
A1 11.86 × 1081.03 × 1074.98 × 104−
A1 25.26 × 1027.60 × 1052.90 × 1043.50 × 105
A1 32.05 × 1022.49 × 1061.67 × 103−
A1 41.13 × 10−2.54 × 1031.01 × 105
A2 11.16 × 1023.57 × 105−−
A2 29.64 × 10−−−
A2 34.95 × 10−9.54 × 102−
A2 47.58 × 10−7.15 × 102−
A2 51.14 ×1022.67 × 1052.02 × 103−
A2 68.49 × 10−−−
A3 17.08 × 108.58 × 1055.52 × 103−
A4 1−1.42 × 106−−
A4 22.61 × 10−3.57 × 103−
A4 31.94 × 10−−−
A4 4−−6.59 × 102−
A4 53.83 × 10−1.80 × 102−
A4 6−−−−
A4 7−1.11 × 106−−
A4 84.55 × 106.59 × 106−−
A4 97.41 × 10−−−
A4 102.68 × 102−−−
A4 112.72 × 102−−−
A4 121.31 × 103−−−
A4 136.96 × 10−−−
Mean7.75 × 106 a1.01 × 106 ab4.03E × 103 ab1.89 × 104 bc
SD3.80 × 1072.43 × 1061.14 × 1047.41 ×104

1 SD – standard deviation. Data represent numbers of virus copies/μL of analyte. Viral loads of some DWV-positive samples could not be calculated because of a lack of viral standards for the helicase-coding region targeted by one of the primer pairs used. – denotes a number of viral copies under the detection limit. Mean viral loads of materials with different lowercase letters are statistically significantly different (P < 0.05)

Table 5.

Viral loads of BQCV and SBV in samples of bees, pollen, honey, hive debris and hive grid smears

BQCVSBV
SampleBeesPollenHoneyDebrisSmearsBeesPollenHoneyDebrisSmears
A1 14.23 × 1071.58 × 1072.83 × 1067.21 × 105−1.76 × 1045.86 × 1061.01 × 1068.39 × 106−
A1 25.05 × 1082.40 × 1063.08 × 1062.10 × 106−1.53 × 1052.00 × 1052.30 × 1066.74 × 106−
A1 35.24 × 1073.28 × 1063.22 × 1061.43 × 105−1.64 × 1052.06 × 1063.81 × 1063.98 × 106−
A1 45.57 × 1072.85 × 1052.15 × 1067.58 × 1061.36 × 1053.10 × 1044.53 × 1041.70 × 1061.04 × 107−
A2 15.66 × 106−1.03 × 106−7.87 × 1053.95 × 104−4.96 × 1054.26 × 1082.27 × 104
A2 23.14 × 1065.00 × 1062.03 × 105−−1.41 × 1091.22 × 1067.19 × 1043.31 × 107−
A2 37.56 × 1065.42 × 1063.78 × 1055.94 × 106−9.00 × 1084.41 × 1062.09 × 1051.08 × 1091.19 × 108
A2 45.66 × 1077.62 × 1052.30 × 106−4.18 × 1052.30 × 1083.00 × 1051.99 × 1061.20 × 1092.46 × 105
A2 52.10 × 1078.70 × 1062.23 × 1061.27 × 1067.98 × 1058.48 × 1091.57 × 1071.79 × 1062.94 × 1094.41 × 106
A2 69.89 × 107−3.59 × 1067.24 × 1057.99 × 1053.06 × 1067.91 × 1041.48 × 1068.68 × 1063.53 × 105
A3 11.40 × 1072.81 × 1064.17 × 1067.70 × 106−4.73 × 1051.08 × 1062.33 × 1065.06 × 1072.14 × 104
A4 13.94 × 1073.71 × 1074.94 × 106−−2.96 × 1058.30 × 1068.07 × 105−−
A4 21.90 × 1082.37 × 1071.75 × 1062.63 × 1062.28 × 1054.20 × 1084.20 × 1061.71 × 1058.60 × 107−
A4 36.86 × 1072.62 × 1071.69 × 105−−1.01 × 1054.21 × 1064.26 × 1041.40 × 107−
A4 44.74 × 1075.59 × 1063.40 × 105−−2.09 × 1051.33 × 1066.47 × 1041.64 × 107−
A4 51.16 × 1081.52 × 1073.57 × 1061.05 × 106−1.22 × 1053.29 × 1066.65 × 1051.08 × 1068.41 × 103
A4 64.93 × 107−4.86 × 106−6.87 × 1042.91 × 105−1.69 × 1065.34 × 106−
A4 71.07 × 1083.50 × 1071.48 × 106−−1.16 × 1054.60 × 1062.38 × 105−−
A4 81.48 × 1088.35 × 1071.10 × 106−5.03 × 1051.12 × 1054.61 × 1061.07 × 105−1.05 × 105
A4 91.62 × 1082.45 × 1077.64 × 104−2.42 × 1056.43 × 1081.95 × 1061.58 × 104−−
A4 109.39 × 1072.40 × 1071.90 × 106−−3.51 × 1052.93 × 1067.64 × 105−−
A4 117.00 × 1072.25 × 1071.16 × 107−−4.20 × 1052.19 × 1074.43 × 106−9.20 × 103
A4 122.82 × 1083.74 × 1073.55 × 105−1.60 × 1054.10 × 1051.10 × 1075.64 × 104−4.56 × 104
A4 131.00 × 1083.32 × 1071.15 × 1065.84 × 1052.59 × 1051.72 × 1098.10 × 1062.88 × 1054.19 × 1087.95 × 105
Mean9.73 × 107 a1.72 × 107 b2.44 × 106 bc1.27 × 106 cd1.83 × 105 d5.75 × 108 a4.47 × 106 a1.11 × 106 a2.63 × 108 a5.21 × 106 b
SD1.09 × 1081.93 × 1072.46 × 1062.37 × 1062.75 × 1051.75 × 1095.38 × 1061.22 × 1066.58 × 1082.43 × 107

1 SD – standard deviation. Data represent numbers of virus copies/μL of analyte. – denotes a number of viral copies in samples under the detection limit. Mean viral loads of materials with different lowercase letters are statistically significantly different (P < 0.05)

DOI: https://doi.org/10.2478/jvetres-2023-0046 | Journal eISSN: 2450-8608 (formerly 2300-3235)
Journal RSS Feed
Language: English
Page range: 323 - 331
Submitted on: Mar 22, 2023
Accepted on: Jul 21, 2023
Published on: Sep 20, 2023
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Keywords:
acute bee paralysis virus,
black queen cell virus,
deformed wing virus,
honey bee viruses,
sacbrood virus
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Life sciences, other,
Medicine,
Veterinary medicine

© 2023 Eliška Čukanová, Jana Prodělalová, Miroslava Palíková, Kristýna Kováčová, Petr Linhart, Ivana Papežíková, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Volume 67 (2023): Issue 3 (September 2023)
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