Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine
Circovirus type 2 are an extremely common swine virus in terms of [is a swine virus extremely widespread in] industrial farming worldwide, including Ukraine]. A similar situation was de-scribed in Ukraine. It leads to an immunosuppressive condition of animals and high mortality of animals. Due to th...
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Інститут молекулярної біології і генетики НАН України
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Dudar, L.V. Budzanivska, I.G. Polishchuk, V.P. 2019-06-15T12:05:16Z 2019-06-15T12:05:16Z 2018 Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine / L.V. Dudar, I.G. Budzanivska, V.P. Polishchuk // Вiopolymers and Cell. — 2018. — Т. 34, № 1. — С. 41-48. — Бібліогр.: 18 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.00096F https://nasplib.isofts.kiev.ua/handle/123456789/154266 578 Circovirus type 2 are an extremely common swine virus in terms of [is a swine virus extremely widespread in] industrial farming worldwide, including Ukraine]. A similar situation was de-scribed in Ukraine. It leads to an immunosuppressive condition of animals and high mortality of animals. Due to the [Considering a] variety of PCV-2 strains in the world, and the difference be-tween strains common in wildlife and farms, [it is necessary to characterize] the characterization of field isolates found among wild boars in Ukraine is required. So, in this work [the] isolates from wild boars from different regions of Ukraine were characterized and differentiated by phy-logenent [phylogenetic] analysis. As a result of the study, it was shown that [the] isolates from the Chernihiv, Zaporizhzhya, Cherkasy and Kharkiv regions belong to different subgroups of the PCV-2 and have different origin. In addition [Additionally], a relatively high level of similarity with the isolates from Croatia and Brazil has been established. At the same time, the similarity of isolates isolated from wild boars from the Zaporizhzhya and Chernihiv oblasts has been shown to have been isolated from pigs from industrial farms. [At the same time, it has been shown the simi-larity of isolates from wild boars from the Zaporizhzhya and Chernihiv regions with those from pigs of industrial farms.] Цирковірус свиней 2 типу є надзвичайно розповсюдженим у всьому світі вірусом серед свиней в умовах промислового вирощування. Ураження вірусом призводить до імуносупресивного стану тварин та високої смертності. З огляду на велику різноманітність штамів ЦВС-2 у світі, та відмінність між штамами, розповсюдженими в дикій природі та на фермах, необхідною є характеристика польових ізолятів, виявлених в Україні серед диких кабанів. Мета. Охарактеризувати та диференціювати ізоляти ЦВС-2, виділені від диких кабанів з різних регіонів України. Методи. Філогенентичний аналіз. Результати. Показано, що ізоляти з Чернігівської, Запорізької, Черкаської та Харківської областей належать до різних субгруп ЦВС-2 та мають різне походження. Крім того, встановлено досить високий рівень їхньої подібності з ізолятами з Хорватії та Бразилії. Водночас, показано подібність ізолятів, виділених від диких кабанів із Запорізької та Чернігівської областей з такими, що були виділені від свиней з промислових господарств. Висновки. Цирковирус свиней 2 типа является чрезвычайно распространенным во всем мире вирусом среди свиней в условиях промышленного выращивания. Заражение вируса приводит к иммуносупрессивному состоянию животных и высокой смертности. Учитывая большое разнообразие штаммов ЦВС-2 в мире, и различие между штаммами, распространенными в дикой природе и на фермах, необходима характеристика полевых изолятов, выявленных в Украине среди диких кабанов. Цель. Охарактеризовать и дифференцировать изоляты ЦВС-2, выделенные от диких кабанов из разных регионов Украины. Методы. Филогенентический анализ . Результаты. Показано, что изоляты из Черниговской, Запорожской, Черкасской и Харьковской областей относятся к различным субгрупам ЦВС-2 и имеют разное происхождение. Кроме того, установлено достаточно высокий уровень их сходства с изолятами из Хорватии и Бразилии. В то же время, показано сходство изолятов, выделенных от диких кабанов Запорожской и Черниговской областей, с выделенными от свиней из промышленных хозяйств. Выводы. Ключевые слова: цирковирус свиней 2 типа, дикие кабаны, филогенетический анализ, генетическое разнообразие. en Інститут молекулярної біології і генетики НАН України Вiopolymers and Cell Viruses and Cell Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine Генетична характеристика ізолятів цирковірусу свиней 2 типу (ЦВС-2), детектованих від диких кабанів з різних регіонів України Генетическая характеристика изолятов цирковируса свиней 2 типа (ЦВС-2), детектированных от диких кабанов с разных регионов Украины Article published earlier |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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DSpace DC |
| title |
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine |
| spellingShingle |
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine Dudar, L.V. Budzanivska, I.G. Polishchuk, V.P. Viruses and Cell |
| title_short |
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine |
| title_full |
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine |
| title_fullStr |
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine |
| title_full_unstemmed |
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine |
| title_sort |
genetic characterization of porcine circovirus type 2 (pcv2) from wild boars detected in different regions of ukraine |
| author |
Dudar, L.V. Budzanivska, I.G. Polishchuk, V.P. |
| author_facet |
Dudar, L.V. Budzanivska, I.G. Polishchuk, V.P. |
| topic |
Viruses and Cell |
| topic_facet |
Viruses and Cell |
| publishDate |
2018 |
| language |
English |
| container_title |
Вiopolymers and Cell |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| format |
Article |
| title_alt |
Генетична характеристика ізолятів цирковірусу свиней 2 типу (ЦВС-2), детектованих від диких кабанів з різних регіонів України Генетическая характеристика изолятов цирковируса свиней 2 типа (ЦВС-2), детектированных от диких кабанов с разных регионов Украины |
| description |
Circovirus type 2 are an extremely common swine virus in terms of [is a swine virus extremely widespread in] industrial farming worldwide, including Ukraine]. A similar situation was de-scribed in Ukraine. It leads to an immunosuppressive condition of animals and high mortality of animals. Due to the [Considering a] variety of PCV-2 strains in the world, and the difference be-tween strains common in wildlife and farms, [it is necessary to characterize] the characterization of field isolates found among wild boars in Ukraine is required. So, in this work [the] isolates from wild boars from different regions of Ukraine were characterized and differentiated by phy-logenent [phylogenetic] analysis. As a result of the study, it was shown that [the] isolates from the Chernihiv, Zaporizhzhya, Cherkasy and Kharkiv regions belong to different subgroups of the PCV-2 and have different origin. In addition [Additionally], a relatively high level of similarity with the isolates from Croatia and Brazil has been established. At the same time, the similarity of isolates isolated from wild boars from the Zaporizhzhya and Chernihiv oblasts has been shown to have been isolated from pigs from industrial farms. [At the same time, it has been shown the simi-larity of isolates from wild boars from the Zaporizhzhya and Chernihiv regions with those from pigs of industrial farms.]
Цирковірус свиней 2 типу є надзвичайно розповсюдженим у всьому світі вірусом серед свиней в умовах промислового вирощування. Ураження вірусом призводить до імуносупресивного стану тварин та високої смертності. З огляду на велику різноманітність штамів ЦВС-2 у світі, та відмінність між штамами, розповсюдженими в дикій природі та на фермах, необхідною є характеристика польових ізолятів, виявлених в Україні серед диких кабанів. Мета. Охарактеризувати та диференціювати ізоляти ЦВС-2, виділені від диких кабанів з різних регіонів України. Методи. Філогенентичний аналіз. Результати. Показано, що ізоляти з Чернігівської, Запорізької, Черкаської та Харківської областей належать до різних субгруп ЦВС-2 та мають різне походження. Крім того, встановлено досить високий рівень їхньої подібності з ізолятами з Хорватії та Бразилії. Водночас, показано подібність ізолятів, виділених від диких кабанів із Запорізької та Чернігівської областей з такими, що були виділені від свиней з промислових господарств. Висновки.
Цирковирус свиней 2 типа является чрезвычайно распространенным во всем мире вирусом среди свиней в условиях промышленного выращивания. Заражение вируса приводит к иммуносупрессивному состоянию животных и высокой смертности. Учитывая большое разнообразие штаммов ЦВС-2 в мире, и различие между штаммами, распространенными в дикой природе и на фермах, необходима характеристика полевых изолятов, выявленных в Украине среди диких кабанов. Цель. Охарактеризовать и дифференцировать изоляты ЦВС-2, выделенные от диких кабанов из разных регионов Украины. Методы. Филогенентический анализ . Результаты. Показано, что изоляты из Черниговской, Запорожской, Черкасской и Харьковской областей относятся к различным субгрупам ЦВС-2 и имеют разное происхождение. Кроме того, установлено достаточно высокий уровень их сходства с изолятами из Хорватии и Бразилии. В то же время, показано сходство изолятов, выделенных от диких кабанов Запорожской и Черниговской областей, с выделенными от свиней из промышленных хозяйств. Выводы. Ключевые слова: цирковирус свиней 2 типа, дикие кабаны, филогенетический анализ, генетическое разнообразие.
|
| issn |
0233-7657 |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/154266 |
| citation_txt |
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine / L.V. Dudar, I.G. Budzanivska, V.P. Polishchuk // Вiopolymers and Cell. — 2018. — Т. 34, № 1. — С. 41-48. — Бібліогр.: 18 назв. — англ. |
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41
L. V. Dudar, I. G. Budzanivska, V. P. Polishchuk
© 2018 L. V. Dudar et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Bio-
polymers and Cell. This is an Open Access article distributed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium,
provided the original work is properly cited
UDС 578
Genetic characterization of porcine circovirus type 2 (PCV2) from
wild boars detected in different regions of Ukraine
L. V. Dudar, I. G. Budzanivska, V. P. Polishchuk
ESC "Institute of Biology and Medicine", Taras Shevchenko National University of Kyiv
64/13, Volodymyrska Str., Kyiv, Ukraine, 01601
liudmyla.dudar@hipra.com
Aim. Circovirus type 2 is a common swine virus in industrial farms worldwide, including
Ukraine. PCV2 infection leads to an immunosuppressive condition and high mortality of
animals. Methods. Due to the variety of PCV-2 strains in the world, and the difference between
wildlife and farm strains, we undertook the characterization of field isolates found among wild
boars in Ukraine using phylogenetic analysis. Results. We have shown that isolates from the
Chernihiv, Zaporizhzhya, Cherkasy and Kharkiv regions belong to different subgroups of the
PCV-2 and have different origins. In addition, a relatively high level of similarity with the
isolates from Croatia and Brazil was found. At the same time, the isolates from wild boars
from the Zaporizhzhya and Chernihiv regions, were found to be similar to that from pigs from
industrial farms. Conclusions. A high level of genetic diversity was found among the inves-
tigated isolates of PCV-2 isolated from wild boars in Ukraine. The fact that some of the isolates
of PCV-2 from wild boars characterized by high level of simillarity with isolates, from pigs
of industrial farms, indicates an existing pathway for the transmission of the virus between
demarcated animal populations and requires further research.
K e y w o r d s: porcine circovirus type 2, wild boar, phylogenetic analysis, genetic diversity.
Introduction
Porcine circovirus type 2 (PCV2) impacts
global swine production, is economically
important, and is associated with multiple
disease entities including multisystemic dis-
ease, wasting, pneumonia, diarrhea and re-
productive failure. Porcine circovirus (PCV),
which belongs to the family Circoviridae,
genus Circovirus [1], is one of the smallest
animal viruses with unenveloped, single-
stranded circular genome and a size of 17 nm
in diameter [2, 3]. Two species of PCV, PCV1
and PCV2, have been characterized [4].
PCV1 is considered to be nonpathogenic to
pigs by experimental inoculation [1], and has
originally been identified as a persistent tis-
sue culture contaminant in pig kidney cell
lines [5]. PCV2 has been shown to be the
causative agent of the post-weaning multi-
ISSN 1993-6842 (on-line); ISSN 0233-7657 (print)
Biopolymers and Cell. 2018. Vol. 34. N 1. P 41–48
doi: http://dx.doi.org/10.7124/bc.00096F
42
L. V. Dudar, I. G. Budzanivska, V. P. Polishchuk
systemic wasting syndrome (PMWS) of
pigs [6, 7].
At least, five Open Reading Frames (ORF)
have been reported to be effectively transcribed,
but the most studied and widely sequenced
region is the ORF2, which encodes for the Cap
protein. This protein represents the only com-
ponent of the viral capsid and has been proven
to be the major target of the host immune re-
sponse. The ORF2-based classification criteria
have been collectively adopted to define the
PCV2 genotypes because of its higher phylo-
genetic signal and lower tendency to recombine.
The PCV2 geographical distribution and its
wide presence have been undoubtedly linked to
livestock movements and trade routes leading
to the rapid spread of new strains in various
countries. The PCV2 genotype 2a is considered
to be prevalent until 2003, when a change in
genotype prevalence (commonly known as a
genotype shift) occurred in favour of the geno-
type 2b, with a parallel enhancement of the
outbreak severity. A similar situation happened
again in 2010 when the genotype 2d appeared
and rapidly spreaded to the detriment of PCV2b
prevalence. The aforementioned high evolution-
ary rate (about 10−3–10−4 substitutions/site/year)
and the huge viral population size provide op-
timal conditions for natural selection to act. One
of the suggested reasons for its wide acquired
genetic variability could reside in a selective
pressure promoted also by the vaccination-in-
duced immunity escape [8].
Consequently, the wild swine population
represents a major source of genetic variabil-
ity and/or simply of viral exchange, but it is
not concerned by the vaccination burden
(Porcine circovirus type 2, PCV2) evolution
before and after the vaccination introduction.
The aim of this study was to bring together
the available genomic information on the wild
and domestic pig PCV2 strains from different
regions of Ukraine to explore their evolution-
ary pathways.
Materials and Methods
Field samples: clinical samples (serum samples
and lymph nodes) from the different farms in
high pig density provinces of Ukraine submitted
to Molecular Diagnostic Laboratory at CVD
(Center of Veterinary Diagnostics) du ring 2014–
2015 were included in this study. These samples
were kept at –80°C until performing DNA ex-
traction and PCR. Viral DNA was extracted
from lymphoid tissue homogenates and serum
samples using NucleoSpin Extract Viral DNA
Kit (Macherey-Nagel, Düren, Germany) accor-
ding to the manufacturer’s instructions.
PCR amplification: A full-length ORF2
gene of PCV2 was amplified in 50 µl of reac-
tion mixture by PCR with forward primer,
PCV2-f1 (5’-CCA TGC CCT GAA TTT CCA
TA-3’) and reverse primer PCV2-r1 (5’-ACA
GCG CAC TTC TTT CGT TT-3’) according
to Takahagi et al. (2008). The amplification
reaction was performed with an initial step at
94°C for 2 min, followed by 35 cycles of de-
naturation at 94°C for 30 s, annealing at 60°C
for 30 s, extension at 72°C for 1 min and a
final extension step at 72°C for 7 min. The
PCV2 positive samples of 702 nt were used
for DNA sequencing.
Sequencing and phylogenetic analysis: the
PCR products were separated by 1.5% agarose
gel electrophoresis and purified with Nucleo-
Spin Extract II (Macherey-Nagel, Düren,
Germany) for the sequences. DNA sequencing
was carried out with primers used in the previ-
43
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine
ous PCR reaction. A total of 4 sequences from
Ukrainian pigs were obtained and translated
into amino acid sequences andanalyzed to-
gether with the representative complete ge-
nome sequences reported in GenBank. A phy-
logenetic tree was constructed by MEGA 6
software (Tamura et al., 2007) using the neigh-
bor-joining (NJ) method with 1000 bootstrap-
ping replicates (Saitou and Nei, 1987).
Results and Discussion
All four PCV2 sequences from wild boars
detected in Ukraine in this study had a genome
length of 1768 nt and revealed nucleotide iden-
tities ranged between 99–91.5% (Tab. 1), in-
dicating no significant difference between
PCV2 genotype of wild boars (complete se-
quences) from different countries.
Ukrainian strains from the Chernigiv,
Cherkassy and Zaporigga regions have a com-
mon origin with the strains from Brazil and
Germany. The strain from Kharkiv has a com-
mon origin with the strain from Croatia that
was allocated in 2009.
The evolutionary history was inferred by
using the Maximum Likelihood method based
on the JTT matrix-based model [9]. The tree
with the highest log likelihood (–3225.2487)
is shown. The percentage of trees in which the
associated taxa clustered together is shown
next to the branches. Initial tree(s) for the
heuristic search were obtained by applying the
BioNJ method to a matrix of pairwise dis-
tances estimated using a JTT model. A discrete
Gamma distribution was used to model evo-
lutionary rate differences among sites (2 ca te-
go ries (+G, parameter = 1.7649)). The tree is
drawn to scale, with branch lengths measured
in the number of substitutions per site. The
analysis involved 17 amino acid sequences.
The coding data were translated assuming a
Standard genetic code table. All positions con-
taining gaps and missing data were eliminated.
There were a total of 539 positions in the final
dataset. Evolutionary analyses were conducted
in MEGA6 [10].
The evolutionary history was inferred by
using the Maximum Likelihood method based
Table 1. Comparison of Ukrainian isolates of PCV2 from wild boar[s] (AK and NT sequences of complete
genome) with PCV2 strains
wb_Chernigiv_UA_16 wb_Cherkasy_UA_16 wb_Charkiv_UA_16 wb_Zaporigga_UA_16
NK AK NK AK NK AK NK AK
P._circovirus_2_BJ0402 98.7 97.2 99 97 96 91.5 99.7 99.4
wild_boar_Germ 95.7 91.1 96 91.3 95.7 90.4 95.9 90.9
wild_boar_Croatia_09 96.2 91.8 96.4 92 99.7 99.1 96.2 91.5
wild_boar_Brazil_12 98.8 97.8 99 98 96.5 92.9 98.6 96.7
wild_boar_72/12_52_Brazil_12 98.5 96.5 98.6 96.3 95.9 90.9 99.4 98.7
wild_boar_72/12_49_Br 98.5 96.5 98.6 96.3 95.9 90.9 99.4 98.7
wild_boar_72/12_36_Br 98.4 96.5 98.5 96.3 95.8 90.9 99.4 98.7
wild_boar_72/12_35_Br 98.5 96.5 98.6 96.3 95.9 90.9 99.4 98.7
wild_boar_72/12_34_Br 98.4 96.3 98.6 96.1 95.8 90.7 99.4 98.5
wild_boar_BRA_JAV2_2008 98.7 97.2 98.8 97 96.1 91.5 99.7 99.4
44
L. V. Dudar, I. G. Budzanivska, V. P. Polishchuk
wild boar 72/12 35
wild boar 72/12 34
wild boar 72/12 36
wild boar 72/12 49
wild boar 72/12 52 Brazil 12
wb Zaporigga UA 16
P. circovirus 2 BJ0402
wild boar BRA JAV2 2008
wild boar BRA JAV1 2008
sw Chernigiv UA 16
wb Chernigiv UA 16
wb Cherkasy UA 16
wild boar Brazil 12
wild boar Germ
wd Kharkiv UA 16
wild boar Croatia 09
PCV1-Hun Sus scrofa
86
88
94
98
86
98
100
39
0.05
Fig. 1. Molecular Phyloge-
netic analysis of PCV2 from
wild boars (AC) by Maxi-
mum Likelihood method.
wild boar 72/12 52 Brazil 12
wild boar 72/12 49
wild boar 72/12 36
wild boar 72/12 35
wild boar 72/12 34
wild boar BRA JAV2 2008
wild boar BRA JAV1 2008
wb Zaporigga UA 16
sw Chernigiv UA 16
P. circovirus 2 BJ0402
wb Chernigiv UA 16
wb Cherkasy UA 16
wild boar Brazil 12
wild boar Germ
wd Kharkiv UA 16
wild boar Croatia 09
PCV1-Hun Sus scrofa
80
42 88
72
84
68
88100
76
100
0.05
Fig. 2. Molecular Phyloge-
netic analysis of PCV2 from
wild boars (NK) by Maxi-
mum Likelihood method.
45
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine
SXTY PCV capsid protein
wild boar BRA JAV2 2008
SXTY14 ORF2
Henan cap protein
AUT5 capsid protein
Fd1 capsid
ZHZ1 cap protein
WB-H-1 capsid protein
DK558control capsid protein
NIVS-5 putative capsid protein
PT-34765-06 cap
CH3 capsid protein
sw Zaporigga UA 16
sw Chernigiv UA 16
wb Zaporigga UA 16
NL Control 3 capsid protein
wild boar 72/12 52
wild boar 72/12 49
wild boar 72/12 36
wild boar 72/12 35
wild boar 72/12 34
DK442case capsid protein
Bj2010PG cap
wb Cherkasy UA 16
PCV2 Uy 1 Cap
wb Chernigiv UA 16
PT-15152-03 cap
C7201-1 capsid protein
TangHe putative capsid protein
NL Control 1 capsid protein
FJFQ0511 cap
YZ cap protein
HN1-5 cap
PT-55935-05 cap
CHST cap
1 A/B
wild boar cap Crov
wd Kharkiv UA 16
09GD Cap
wild boar Cap Germ
sw Cherkasy UA 16
sw Kharkiv UA 16
2
91
97
62
72
88
93
72
60
24
58
97
91
58
93
66
34
0.01
Fig..3. Evolutionary relationships of
PCV2 from wild boars and pigs
46
L. V. Dudar, I. G. Budzanivska, V. P. Polishchuk
on the Kimura 2-parameter model [11]. The
tree with the highest log likelihood
(–4739.0701) is shown. The percentage of
trees in which the associated taxa clustered
together is shown next to the branches. Initial
tree(s) for the heuristic search were obtained
by applying the BioNJ method to a matrix of
pairwise distances estimated using the
Maximum Composite Like lihood (MCL) ap-
proach. A discrete Gamma distribution was
used to mo del evolutionary rate differences
among sites (2 categories (+G, parameter =
0.0500)). The tree is drawn to scale, with
branch lengths measured in the number of
substitutions per site. The analysis involved
17 nucleotide sequences. Codon positions in-
cluded were 1st+2nd+3rd. All positions con-
taining gaps and missing data were eliminated.
There were a total of 1743 positions in the
final dataset. Evolutionary analyses were con-
ducted in MEGA6 [10].
The tree topology based on the nucleotide and
amino acid sequence data coincides which con-
firms the validity of our conclusions (Fig.1, 2).
Comparison of the circulating strains of
wild and domestic animals in Ukraine showed
that most of them are quite different and be-
long to different subgroups. The same situation
was described in other European countries.
However, the strains isolated from the pigs of
the Zaporigga and Chernigiv regions have a
common origin with the strain isolated from
wild boars in the Zaporigga region. The strains
isolated from the pigs from the Cherkasy and
Kharkiv regions have a common origin with
the strain isolated from wild boars from the
Kharkiv region (Fig..3). This fact demonstrates
a common origin of described isolates and
could be caused by direct transmission of
PCV-2 between wild boars and farm animals,
that is not allowed by the biosecurity policy.
The evolutionary history was inferred using
the Neighbor-Joining method [12]. The opti-
mal tree with the sum of branch length =
0.28887349 is shown. The percentage of rep-
licate trees in which the associated taxa clus-
tered together in the bootstrap test (1000 rep-
licates) is shown next to the branches [13]. The
tree is drawn to scale, with branch lengths in
the same units as those of the evolutionary
distances used to infer the phylogenetic tree.
The evolutionary distances were computed
using the p-distance method and are in the
units of the number of amino acid differences
per site. The analysis involved 41 amino acid
sequences. The coding data were translated
assuming a Standard genetic code table. All
positions containing gaps and missing data
were eliminated. There were a total of 124 po-
sitions in the final dataset. Evolutionary anal-
yses were conducted in MEGA6 [10].
The phylogenetic tree analyses classified the
isolates of this study in two subgroups accord-
ing to the classification proposed by Grau-Roma
et al. (2008) [14]. Based on the subgroup ter-
minology described previously (Olvera et al.,
2007) [15], nucleotides 262–267 and amino
acids 88–89 of ORF2 were compared and clas-
sified. The nucleotide sequen ces “CCCCGC”,
“CCCCTC” and “AAAATC” are the signatures
motif for PCV2b subgroup 1A/B, 1C and
PCV2a, respectively. The amino acid “PR” was
enclosed with subgroup1A/B, while the PL and
KI were related with subgroup 1C and PCV2a
(Cheung et al., 2007) [16]. The isolates from
Zaporigga, Chernigiv and Cherkasy 1 were
divided into 1A/ B subgroups together with
PCV2 sequences of strains from wild boars
47
Genetic characterization of porcine circovirus type 2 (PCV2) from wild boars detected in different regions of Ukraine
from Brazil and Germany. The isolates from
Charkiv 1 were divided into 2 subgroups to-
gether with PCV2 sequences of strain from wild
boars from Croatia (Fig.1–3).
Genetically, the characteristics of Ukrainian
isolates of PCV2 showed a high level of di-
versity. As described, the detected isolates
belong to both subgroups (1A\B and 2) of PCV
strains. Additionally to the interesting model
that could be applied to other human and ani-
mal diseases, this rouses the interest to a con-
tinuous monitoring of viral epidemiology, par-
ticularly for rapidly evolving viruses like
PCV2, and the necessity to share the related
information to prevent or promptly act in re-
sponse to the potential emergence of actual
vaccine-immunity escape mutants [17–19].
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Генетична характеристика ізолятів цирковірусу
свиней 2 типу (ЦВС-2), детектованих від диких
кабанів з різних регіонів України
Л. В. Дудар, І. Г. Будзанівська, В. П. Поліщук
Цирковірус свиней 2 типу є надзвичайно розповсю-
дженим у всьому світі вірусом серед свиней в умовах
промислового вирощування. Ураження вірусом при-
зводить до імуносупресивного стану тварин та високої
смертності. З огляду на велику різноманітність штамів
ЦВС-2 у світі, та відмінність між штамами, розпо-
всюдженими в дикій природі та на фермах, необхідною
є характеристика польових ізолятів, виявлених в
Україні серед диких кабанів. Мета. Охарактеризувати
та диференціювати ізоляти ЦВС-2, виділені від диких
кабанів з різних регіонів України. Методи. Філо ге нен-
тич ний аналіз. Результати. Показано, що ізоляти з
Чернігівської, Запорізької, Черкаської та Харківської
областей належать до різних субгруп ЦВС-2 та мають
різне походження. Крім того, встановлено досить ви-
сокий рівень їхньої подібності з ізолятами з Хорватії
та Бразилії. Водночас, показано подібність ізолятів,
виділених від диких кабанів із Запорізької та Чер ні-
гівсь кої областей з такими, що були виділені від свиней
з промислових господарств. Висновки. Високий рівень
генетичного різноманіття був виявлений серед дослі-
джених ізолятів ЦВС-2, виділених від диких кабанів
України. Установлений факт подібності деяких ізолятів
ЦВС-2 від диких кабанів з ізолятами, від свиней з
промислових ферм, свідчить про існуючий шлях пе-
редачі вірусу між розмежованими популяціями тварин
і потребує подальших досліджень.
К л юч ов і с л ов а: цирковірус свиней 2 типу, дикі
кабани, філогенетичний аналіз, генетичне різноманіття.
Генетическая характеристика изолятов
цирковируса свиней 2 типа (ЦВС-2),
детектированных от диких кабанов с разных
регионов Украины
Л. В. Дудар, И. Г. Будзанивская, В. П. Полищук
Цирковирус свиней 2 типа является чрезвычайно рас-
пространенным во всем мире вирусом среди свиней в
условиях промышленного выращивания. Заражение
вируса приводит к иммуносупрессивному состоянию
животных и высокой смертности. Учитывая большое
разнообразие штаммов ЦВС-2 в мире, и различие
между штаммами, распространенными в дикой при-
роде и на фермах, необходима характеристика полевых
изолятов, выявленных в Украине среди диких кабанов.
Цель. Охарактеризовать и дифференцировать изоляты
ЦВС-2, выделенные от диких кабанов из разных ре-
гионов Украины. Методы. Филогенентический анализ.
Результаты. Показано, что изоляты из Черниговской,
Запорожской, Черкасской и Харьковской областей
относятся к различным субгрупам ЦВС-2 и имеют
разное происхождение. Кроме того, установлено до-
статочно высокий уровень их сходства с изолятами из
Хорватии и Бразилии. В то же время, показано сход-
ство изолятов, выделенных от диких кабанов Запо рож-
ской и Черниговской областей, с выделенными от
свиней из промышленных хозяйств. Выводы. Высокий
уровень генетического разнообразия был выявлен
среди исследованных изолятов ЦВС-2, выделенных от
диких кабанов Украины. Установленный факт родства
некоторых изолятов ЦВС-2 от диких кабанов с изоля-
тами, от свиней из промышленных свиноводческих
ферм, указывает на существующий путь передачи
вируса между разграниченными популяциями живот-
ных и требует дальнейших исследований.
К л юч е в ы е с л ов а: цирковирус свиней 2 типа,
дикие кабаны, филогенетический анализ, генетическое
разнообразие.
Received 22.11.2017
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