Species of the genus Varroa are ectoparasitic mites of the Apis honey bees. Unlike the well-known species of mites V. destructor and V. jacobsoni, V. underwoodi is still poorly studied. According to foreign publications, the currently recognized distribution of V. underwoodi in the A. cerana population includes Nepal, South Korea, Japan, Malaysia, India, Indonesia, Papua New Guinea, Vietnam, and China. Recently it has been detected in Russia (Primorsky krai) on A. cerana ussuriensis honey bees. Since V. destructor and V. jacobsoni have the ability to easily switch to other bee species, there is a possibility that later V. underwoodi may also extensively switch from the Asian honey bees A. cerana to the European honey bees A. mellifera. The first case of V. underwoodi parasitizing in A. mellifera colonies was recorded in Papua New Guinea. The parasitic mite V. underwoodi requires careful study, since it is a new potential parasite of the honey bees A. mellifera, which can also bring new species and strains of viruses and bacteria, change the composition of the gut microbiome, and disrupt the protective and adaptive mechanisms of the bees. This article presents the data of morphometry and polymorphism of the COX1 gene mtDNA. Varroa underwoodi was compared with the other mite species V. destructor and V. jacobsoni. The mean genetic divergence and p-distance between V. underwoodi and other Varroa species were 9% and 0.09, respectively, which is consistent with the level of species differences in insects. The nucleotide sequences of the COX1 gene mtDNA of V. underwoodi from Primorsky krai of Russia LC532104 and from the Jilin province of China MH205176 turned out to be identical and were assigned to the China 1 MH205176 haplotype. It is assumed that there is continuous migration between the A. cerana populations of Russia and China, which led to the spread of V. underwoodi in the natural population of A. cerana ussuriensis in the Primorsky krai of Russia, and the mite is currently distributed up to 45.06° N. Thus, the northern border of the V. underwoodi range is located on the territory of the Russian Far East and, probably, coincides with the range of A. cerana ussuriensis. It is likely that the selection of A. mellifera bee colonies for hygienic behavior against the V. destructor mite may also be effective against V. underwoodi and will prevent the possible transition of V. underwoodi from Asian A. cerana to European A. mellifera.

Ильясов, Р.А., Такахаши, Д.И., Ли, М.Л., Прощалыкин, М.Ю., Лелей, А.С., Квон, Х.В., Даниленко, В.Н., Николенко, А.Г., 2022. Характеристика клещей Varroa underwoodi (Acari Varroidae) в популяции Apis cerana ussuriensis (Hymenoptera: Apidae) в Приморском крае, Россия. Журнал Общей Биологии. 83 (1), 38-50. doi: 10.31857/S0044459622010055. (SCI, WoS, Scopus). IF = 1,028. Ilyasov, R.A., Takahashi, J.I., Lee, M.L., Proshchalykin, M.Y., Lelej, A.S., Kwon, H.W., Danilenko, V.N., Nikolenko, A.G., 2022. Characteristics of Varroa underwoodi mites (Acari: Varroidae) in the population of Apis cerana ussuriensis (Hymenoptera: Apidae) in the Primorsky Krai of Russia. Biology Bulletin Reviews (Zhurnal obshchei biologii). 83 (1), 38-50. doi: 10.31857/S0044459622010055. (SCI, WoS, Scopus). IF = 1,028. ISSN 0044-4596 Abstract. Species of the genus Varroa are ectoparasitic mites of the Apis honey bees. Unlike the well-known species ofmites V. destructor and V. jacobsoni, V. underwoodi is still poorly studied. According to foreign publications, thecurrently recognized distribution of V. underwoodi in the A. cerana population includes Nepal, South Korea,Japan, Malaysia, India, Indonesia, Papua New Guinea, Vietnam, and China. Recently it was discovered inthe Russia (Primorsky Krai) on the honey bees A. cerana ussuriensis. Since V. destructor and V. jacobsoni havethe ability to easily switch to other bee species, there is a possibility that later V. underwoodi may also extensively switch from Asian honey bees A. cerana to European honey bees A. mellifera. The first case of V. underwoodi parasitizing in A. mellifera colonies was recorded in Papua New Guinea. The parasitic mite V. underwoodi requires careful study, since it is a new potential parasite of honey bees A. mellifera, which can also bringnew species and strains of viruses and bacteria, change the composition of the gut microbiome, and disruptthe protective and adaptive mechanisms of the bees. The article presents the data on morphometry and polymorphism of the gene COX1 of mtDNA. Varroa underwoodi was compared with other mite species V. destructor and V. jacobsoni. The mean genetic divergence and p-distance between V. underwoodi and other Varroaspecies were 9% and 0.09, respectively, which are consistent with the level of species differences in insects.The nucleotide sequences of the gene COX1 of mtDNA of V. underwoodi from Primorsky Krai of RussiaLC532104 and from the Jilin province of China MH205176 turned out to be identical and were assigned tothe China 1 MH205176 haplotype. It is assumed that there is continuous migration between the A. ceranapopulations of Russia and China, which led to the spread of V. underwoodi in the natural population of A. cerana ussuriensis in the Primorsky Krai of Russia, and the mite is currently distributed up to 45.06°N. Thus, thenorthern border of the V. underwoodi range is located on the territory of Russian Far East and, probably, coincides with the range of A. cerana ussuriensis. It is likely that the selection of A. mellifera bee colonies for hygienic behavior against the V. destructor mite may also be effective against V. underwoodi and will prevent thepossible transition of V. underwoodi from Asian A. cerana to European A. mellifera. Аннотация. Виды рода Varroa являются эктопаразитическими клещами медоносных пчел рода Apis. В отличие от хорошо известных видов клещей V. destructor и V. jacobsoni, V. underwoodi остается мало изученным. Современный ареал V. underwoodi в популяции A. cerana включает Непал, Южную Корею, Японию, Малайзию, Индию, Индонезию, Папуа-Новую Гвинею, Вьетнам и Китай. Недавно он был найден в России (Приморский край) на A. cerana ussuriensis. Поскольку V. destructor и V. jacobsoni обладают способностью легко переходить на другие виды медоносных пчел, есть большая вероятность того,что V. underwoodi может в дальнейшем перейти в массовом порядке с A. cerana на A. mellifera. Первый случай паразитирования V. underwoodi в семьях пчел A. mellifera зафиксирован в Папуа-Новой Гвинее. Varroa underwoodi требует тщательного изучения, поскольку является новым потенциальным паразитом A. mellifera и способен принести с собой новые виды и штаммы вирусов и бактерий, изменить состав микробиома кишечника пчел, нарушить защитные и адаптивные механизмы их организма. В статье представлены данные морфометрии V. underwoodi и полиморфизма его гена COX1мтДНК. Проведено сравнение V. underwoodi с другими видами клещей V. destructor и V. jacobsoni. Средняя генетическая дивергенция и p-дистанция между V. underwoodi и другими видами Varroa составляли 9% и 0.09 соответственно, что согласуется со средним уровнем видовых различий у насекомых. Нуклеотидные последовательности гена COX1 мтДНК V. underwoodi из Приморского края (Россия) LC532104 и провинции Цзилинь (Китай) MH205176 оказались идентичными и отнесены кгаплотипу China 1 MH205176. Предполагается, что между популяциями A. cerana России и Китая происходит непрерывный обмен, который привел к появлению V. underwoodi в природной популяции A. cerana ussuriensis в Приморском крае. Современная северная граница ареала V. underwoodi проходит по территории Дальнего Востока России и, вероятно, совпадает с таковой A. cerana ussuriensis (45.06° с.ш.). Вероятно, селекция семей пчел A. mellifera по гигиеническому поведению против клещей V. destructor может оказаться эффективной также против V. underwoodi и позволит предотвратить возможный переход V. underwoodi с азиатских пчел A. cerana на европейских пчел A. mellifera.

Ilyasov, R.A., Takahashi, J.I., Proshchalykin, M.Y., Lelej, A.S., Lee, M.L., Kwon, H.W., Nikolenko, A. G. (2021). First evidence of presence of Varroa underwoodi mites on native Apis cerana colonies in Primorsky Territory of Russia based on COX1 gene. Journal of Apicultural Science 65 (1), 177-186. doi: 10.2478/JAS-2021-0014. Abstract. The species of genus Varroa mites parasitize on the honey bees of genus Apis. Unlike the well-studied V. destructor and V. jacobsoni mites, V. underwoodi remain less known. According to English language publications, the proven V. underwoodi distribution area of A. cerana colonies covers Nepal, South Korea, Indonesia, Papua New Guinea, Vietnam and China, but not Russia even though it had been described morphometrically in Russian language publications in Russia’s Primorsky Territory. According to Vavilov’s law (1920) of a homologous series, all the species of V. underwoodi, V. destructor and V. jacobsoni have the ability to spill over onto new hosts. Thus, V. underwoodi is a potential parasite of A. mellifera that should be carefully studied. In this study, V. underwoodi mites in colonies of honey bee subspecies A. c. ussuriensis native to Russia’s Primorsky Territory are first proven using both morphometry and mitochondrial COX1 gene sequencing. The genetic divergence and p-distances between V. underwoodi and other Varroa species ranged from 7 to 10% and from 0.072 to 0.099, respectively, which matched the intraspecific level of differences. Two identical northernmost V. underwoodi samples from Russia’s Primorsky Territory and China’s Jilin province with GenBank accession number MH205176 were assigned as COX1 haplotype China 1 MH205176. The first discovery of V. underwoodi in the Primorsky Territory in northern Asia outlined the northern border of its range.

Apis cerana ussuriensis Ilyasov et al., 2019 is the northernmost subspecies of the Asian honey bee A. cerana Fabricius, 1793, common in the forests of Primorsky krai and Khabarovsk krai as far as 47°54′ N. Genetic studies of this subspecies are of great interest for science and apiculture, since all its adaptive traits were formed under the influence of the natural environment without human interference. We sequenced and annotated the complete mitochondrial DNA (mtDNA) sequences of bees of subspecies Apis cerana ussurien-sis Ilyasov et al., 2019 (Genbank accession number AP018450) from Primorsky krai and Apis cerana koreana Ilyasov et al., 2019 from South Korea, as well as six exons of the nuclear DNA (nDNA) vitellogenin VG E2

Ильясов Р.А., Хан Г.Ю., Ли М.Л., Ким К.В., Парк Д.Х., Такахаши Д.И., Квон Х.В., Николенко А.Г. Эволюционные взаимоотношения кавказской и карпатской популяций медоносной пчелы. Часть 1. Пчеловодство, 3, 2021. С. 16-19. (Ilyasov R.A., Han G.Y., Lee M.L., Kim K.W., Park J.H., Takahashi J.I., Kwon H.W., Nikolenko A.G. Evolutionary relationships of caucasian Apis mellifera caucasia and carpathian Apis mellifera carpathica poplations of the honey bee. Part 1. Russian Journal of Beekeeping "Pchelovodstvo", Issue 3, 2021. P. 16-19.) Впервые секвенированы оследовательности полного митохондриального генома подвидов пчел Apis mellifera caucasia Pollmann, 1889 (AP018404, 16341 п.н.) и Apis mellifera carpathica Foti et al., 1965 (AP018403, 16336 п.н.). Митохондриальные ДНК (мтДНК) обоих подвидов содержат 13 кодирующих белок генов, 22 гена тРНК, 2 гена рРНК и AT-обогащенную регуляторную область. Отношение транзиций к трансверсиям tr/tv полной мтДНК между A. m. caucasia и A. m. carpathica соответствует 2,05, что характеризует формирование адаптаций к сменяющимся условиям среды обитания. Гены с наибольшим содержанием GC — COX1, COX2, CYTB, COX3 и ND1 могут быть высокополиморфны и использованы в филогенетических и популяционных исследованиях пчел. Большинство генов мтДНК обоих подвидов расположены на тяжелой цепи и меньшее количество — на легкой цепи. Кластерный анализ последовательности полной мтДНК и оценка структуры межгенной области тРНК-Leu(UUR)-COX2 с единственным элементом Q размером 192 п.н. показали, что A. m. caucasia и A. m. carpathica являются представителями линии C с гаплотипами C2 и C2j соответственно. Указанные подвиды можно дифференцировать друг от друга по 34 уникальным SNP в 11 генах мтДНК и маркеру рестрикции XbaI в гене ND5. Эти генетические маркеры могут способствовать сохранению чистопородных генофондов A. m. caucasia и A. m. carpathica в пределах их естественного ареала. Ключевые слова: Apis mellifera, подвиды пчел, A. m. caucasia, A. m. carpathica, митохондриальный геном, мтДНК, гаплотипы, консервативная генетика. The sequences of the complete mitochondrial genome of the honey bee subspecies Apis mellifera caucasia Pollmann, 1889 (AP018404, 16341 bp) and Apis mellifera carpathica Foti et al., 1965 (AP018403, 16336 bp) were first sequenced. Mitochondrial DNA (mtDNA) of both subspecies contains 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and 1 AT-rich regulatory region. The ratio of transitions to transversions tr/tv in complete mtDNA between A. m. caucasia and A. m. carpathica was 2.05, which characterizes the formation of adaptations to changing environmental conditions. Genes with the highest GC content — COX1 (24%), COX2 (19.6%), CYTB (19.1%), COX3 (17.2%) and ND1 (17.2%) can be highly polymorphic and used in phylogenetic and population studies of bees. Most of mtDNA genes for both subspecies are located on the heavy chain (9 protein coding genes and 14 tRNA genes) and fewer genes (4 protein coding genes, 2 rRNA genes and 8 tRNA genes) are located on the light chain. Cluster analysis of the complete mtDNA sequence and assessment of the structure of the tRNA-Leu(UUR)-COX2 intergenic region with a single Q element of 192 bp showed that both subspecies A. m. caucasia and A. m. carpathica are representatives of the line C with haplotypes C2 and C2j, respectively. Subspecies of the honey bee A. m. caucasia and A. m. carpathica can be differentiated from each other by 34 unique SNPs in 11 mtDNA genes and the XbaI restriction marker in the ND5 gene. These genetic markers can contribute to the preservation of purebred gene pools of honey bee subspecies A. m. caucasia and A. m. carpathica within their natural range. Keywords: Apis mellifera, honeybee subspecies, A. m. caucasia, A. m. carpathica, mitochondrial genome, mtDNA, haplotypes, conservation genetics.