Molecular Detection of Nosema spp. in Honey in Bulgaria
Abstract
:1. Introduction
2. Materials and Methods
2.1. Honey Samples
2.2. DNA Extraction for Identification of eDNA from V. destructor, N. apis, and N. ceranae in Honey Samples, Using CTAB Protocol with a Spin Column
2.3. PCR Analysis
2.4. Sequence and Phylogenetic Analysis
3. Results
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Species | Primer Name | Primer Sequences (5′-3′) | Size in bp | Amplified Region | Reference |
---|---|---|---|---|---|
Apis mellifera | AM_Forward AM_Reverse | GGCAGAATAAGTGCATTG TTAATATGAATTAAGTGGGG | C lineage 85 | mtDNA coxI-coxII | [33] |
Varroa destructor | 10KbCOIF1 6,5KbCOIR | CTTGTAATCATAAGGATATTGGAAC AATACCAGTGGGAACCGC | 929 | coxI | [34,35] |
Varroa destructor | 10KbCytbF-1 10KbCytbPRIM | GCAGCTTTAGTGGATTTACCTAC CTACAGGACACGATCCCAAG | 985 | cytb | [34,35] |
Nosemaapis | 321APIS-FOR 321APIS-REV | GGGGGCATGTCTTTGACGTACTATGTA GGGGGGCGTTTAAAATGGAAACAACTATG | 321 | SSU 16S rDNA | [36] |
Nosemaceranae | 218MITOC-FOR 218MITOC-REV | CGGCGACGATGTGATATGAAAATATTAA CCCGGTCATTCTCAAACAAAAAACCG | 218 or 219 a | SSU 16S rDNA | [36] |
N. ceranae GenBank Acc. no. | 6 | 6 | 6 | 6 | 6 | 6 | 6 |
1 | 1 | 1 | 1 | 1 | 2 | 2 | |
8 | 8 | 8 | 8 | 9 | 0 | 0 | |
6 | 6 | 7 | 8 | 6 | 2 | 3 | |
2 | 7 | 7 | 4 | 3 | 9 | 5 | |
NW 020169312 reference | A | A | A | A | A | T | A |
MG657260 Bulgaria | |||||||
MW600361 Mexico | T | ||||||
MN649205 UK | A | ||||||
KC680638 Morocco | |||||||
KC680637 Lebanon | |||||||
MZ044978 Egypt | |||||||
MW396669 Turkey | |||||||
FJ227957 Argentina | |||||||
DQ673615 Switzerland | |||||||
DQ374656 Germany | |||||||
DQ374655 France | |||||||
LC493173 Japan | C | ||||||
LC510232 Japan NCS48 | G | ||||||
LC510233 Japan NCS49 | G | ||||||
HM581509 Mexico | G | ||||||
LC510243 Japan NCS59 | G | ||||||
HM859897 Italy | G | ||||||
HM802210 Mexico | G |
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Salkova, D.; Shumkova, R.; Balkanska, R.; Palova, N.; Neov, B.; Radoslavov, G.; Hristov, P. Molecular Detection of Nosema spp. in Honey in Bulgaria. Vet. Sci. 2022, 9, 10. https://doi.org/10.3390/vetsci9010010
Salkova D, Shumkova R, Balkanska R, Palova N, Neov B, Radoslavov G, Hristov P. Molecular Detection of Nosema spp. in Honey in Bulgaria. Veterinary Sciences. 2022; 9(1):10. https://doi.org/10.3390/vetsci9010010
Chicago/Turabian StyleSalkova, Delka, Rositsa Shumkova, Ralitsa Balkanska, Nadezhda Palova, Boyko Neov, Georgi Radoslavov, and Peter Hristov. 2022. "Molecular Detection of Nosema spp. in Honey in Bulgaria" Veterinary Sciences 9, no. 1: 10. https://doi.org/10.3390/vetsci9010010