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Effect of serial in vivo passages on the adaptation of H1N1 avian influenza virus to pigs

Kinga Urbaniak
Kinga Urbaniak
, 
Andrzej Kowalczyk
Andrzej Kowalczyk
, 
Małgorzata Pomorska-Mól
Małgorzata Pomorska-Mól
, 
Krzysztof Kwit
Krzysztof Kwit
 and 
Iwona Markowska-Daniel
Iwona Markowska-Daniel
   | Mar 25, 2022
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Published Online: Mar 25, 2022
Page range: 9 - 19
Received: Jan 12, 2022
Accepted: Mar 02, 2022
DOI: https://doi.org/10.2478/jvetres-2022-0013
Keywords
© 2022 K. Urbaniak et al. published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

A schematic representation of the experimental timeline. (a) Scheme of virus passages; (b) Scheme of the introduction of contact piglets to experimentally inoculated animals. In each passage/contact group, two piglets were used. p – passage; HI – haemagglutination inhibition; rRT-PCR – real time reverse transcriptase PCR; inocul. – inoculation; i.n. – intranasally; r.t. – rectal temperature; n.s. – nasal swabs; N – passage number; EI – experimentally inoculated; CE – contact-exposed
A schematic representation of the experimental timeline. (a) Scheme of virus passages; (b) Scheme of the introduction of contact piglets to experimentally inoculated animals. In each passage/contact group, two piglets were used. p – passage; HI – haemagglutination inhibition; rRT-PCR – real time reverse transcriptase PCR; inocul. – inoculation; i.n. – intranasally; r.t. – rectal temperature; n.s. – nasal swabs; N – passage number; EI – experimentally inoculated; CE – contact-exposed

Fig. 2

Rectal temperature of experimentally inoculated (EI) and contact-exposed (CE) piglets in individual passages. (a) EI gilts; (b) EI barrows; (c) CE gilts; (d) CE barrows; dpi – days post inoculation; dpc – days post contact; I – first group; II – second group
Rectal temperature of experimentally inoculated (EI) and contact-exposed (CE) piglets in individual passages. (a) EI gilts; (b) EI barrows; (c) CE gilts; (d) CE barrows; dpi – days post inoculation; dpc – days post contact; I – first group; II – second group

Fig. 3

Pathological changes in the lungs of animals from selected passages. (a) experimentally inoculated (EI) gilt at passage (p) 10, lung lesion score (LLS) = 2; (b) EI gilt at p12, LLS = 3; (c) EI gilt at p24, LLS = 8; (d) EI barrow at p24, LLS = 9; (e) contact-exposed (CE) group I gilt at p24, LLS = 11; (f) CE group I barrow at p24, LLS = 6
Pathological changes in the lungs of animals from selected passages. (a) experimentally inoculated (EI) gilt at passage (p) 10, lung lesion score (LLS) = 2; (b) EI gilt at p12, LLS = 3; (c) EI gilt at p24, LLS = 8; (d) EI barrow at p24, LLS = 9; (e) contact-exposed (CE) group I gilt at p24, LLS = 11; (f) CE group I barrow at p24, LLS = 6

Fig. 4

rRT-PCR results. (a) results for all tested tissue samples from EI piglets in each passage; (b) results for all tested tissue samples from CE piglets. p – passage; CE – contact-exposed; b – barrow; g – gilt; I – first group; II – second group
rRT-PCR results. (a) results for all tested tissue samples from EI piglets in each passage; (b) results for all tested tissue samples from CE piglets. p – passage; CE – contact-exposed; b – barrow; g – gilt; I – first group; II – second group

Fig. 5

rRT-PCR results for nasal swabs from the EI and CE piglets. (a) results from animals at passage 20; (b) results from animals at passage 24. p – passage; EI – experimentally inoculated; CE – contact-exposed; b – barrow; g – gilt; I – first group; II – second group; dpi – day post inoculation; dpc – day post contact
rRT-PCR results for nasal swabs from the EI and CE piglets. (a) results from animals at passage 20; (b) results from animals at passage 24. p – passage; EI – experimentally inoculated; CE – contact-exposed; b – barrow; g – gilt; I – first group; II – second group; dpi – day post inoculation; dpc – day post contact

Fig. 6

rRT-PCR results for each tissue type from EI and CE piglets. NM – nasal mucosa; T – trachea; RCrL – right cranial lobe; RML – right middle lobe; RCaL – right caudal lobe; LCrL – left cranial lobe; LML – left middle lobe; LCaL – left caudal lobe; AL – accessory lobe; strong positive Ct < 30; weak positive Ct 30–35; negative Ct > 35
rRT-PCR results for each tissue type from EI and CE piglets. NM – nasal mucosa; T – trachea; RCrL – right cranial lobe; RML – right middle lobe; RCaL – right caudal lobe; LCrL – left cranial lobe; LML – left middle lobe; LCaL – left caudal lobe; AL – accessory lobe; strong positive Ct < 30; weak positive Ct 30–35; negative Ct > 35

Amino acid changes in the proteins of virus strains obtained from selected passages

Gene a.a. position Inoculum vEIp9 vEIp16 vEIp20 vCEp20 vEIp24 vCEp24
144 R Q Q/* * § * §
PB2 477 R G G G § G §
554 I L L L § L §
701 D N N N § N §
PB1 654 S * * G § G §
370 L * * */I § * §
PA 465 I */T * * § * §
613 E * */K */K § */K K
159 S * */R * § * §
180 K E E E § E/G §
189 E * * * § */D §
203 D/N N N N § N §
HA 204 E D D D § D §
239 G * */E E § D §
286 D * */E * § * §
411 G S S S § S §
NP 63 I M M M § M §
132 R K K K § K §
30 I * * L § L §
34 V * * */I I */I §
95 G S S S § S §
NA 127 L M M M § M §
131 A T T T § T §
135 T * * * § * S
364 N S S S § S §
M1 248 M I I I § I §
M2 19 C Y Y Y § Y §
NS 56 M */I I I § I §
226 V I I I § I §

Results of CE piglets’ serological tests

Test dpc p6 p9 p11 p16 p20 p21 p25
g b g b g b g b g b gI bI gII bII
HI 7 < < < < < < < < < < < < < <
14 < < < < < < < < < < n.t. n.t. < <
21 < < < < < < < < < < n.t. n.t. < <
IPMA 7 < < < < < < < < 32 128 64 64 128 64
14 < < < < < < < < 32 256 n.t. n.t. 128 64
21 < < < < < < < < 32 128 n.t. n.t. 64 128
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