Molecular Characterization and Pathogenicity of Chicken Parvovirus (ChPV) in Specific Pathogen-Free Chicks Infected Experimentally
Abstract
:1. Introduction
2. Results
2.1. Molecular Characterization of ChPV
2.2. Experimental Infection—Clinical Signs
2.3. Macroscopic Examination
2.4. Histopathology Examination
2.5. Detection and Quantification of Chicken Parvovirus Genome Copies in the Different Tissues
3. Discussion
4. Materials and Methods
4.1. Virus Strain, Suspension and DNA Extraction
4.2. Sequencing of the VP1 Gene of ChPV for Molecular Characterization
4.3. Experimental Infection
4.4. Obtaining Organs for Detection of ChPV through qPCR
4.5. Macroscopy and Histopathology Examination
4.6. Detection and Quantification of Chicken Parvovirus in Different Tissues through qPCR
4.7. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Number | Sequences | Nucleotides (%) | ||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Numbers of ChPV Sequences | Numbers of TuPV Sequences | |||||||||||||||||
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | ||||
ChPV | 1 | KJ486490.1 Korea | - | 99.6 | 99.8 | 98.8 | 95 | 95 | 92.3 | 91.9 | 92 | 91.1 | 88.3 | 73.5 | 73.3 | 73.5 | 72.9 | |
2 | KJ486489.1 Korea | 99.8 | - | 99.8 | 98.8 | 95.1 | 95.1 | 92.4 | 92.2 | 92.2 | 91.4 | 88.3 | 73.6 | 73.5 | 73.7 | 73.1 | ||
3 | KJ486491.1 Korea | 99.8 | 100 | - | 98.9 | 95.1 | 95 | 92.3 | 92.1 | 92.1 | 91.2 | 88.4 | 73.6 | 73.4 | 73.7 | 73.1 | ||
4 | KM254172.1 USA | 99.8 | 100 | 100 | - | 95 | 94.8 | 93 | 92 | 91.8 | 91.7 | 88.3 | 73.7 | 73.5 | 73.7 | 73.1 | ||
5 | USP362-3 Brazil | 98 | 98.2 | 98.2 | 98.2 | - | 94.9 | 91.8 | 91.8 | 91.8 | 91.6 | 88.2 | 73.6 | 73.4 | 73.6 | 72.9 | ||
6 | KM598414.1 USA | 98.3 | 98.5 | 98.5 | 98.5 | 97.9 | - | 91.3 | 92.7 | 92.2 | 91.9 | 88.7 | 73.2 | 73.2 | 73.4 | 72.8 | ||
7 | KM254173.1 USA | 96.4 | 96.5 | 96.5 | 96.5 | 96.2 | 95.8 | - | 89.5 | 89.2 | 90.7 | 87.7 | 73.1 | 72.9 | 73.2 | 72.6 | ||
8 | KU523900.1 China | 96.4 | 96.5 | 96.5 | 96.5 | 96.7 | 96.7 | 94.8 | - | 91.7 | 92.4 | 89 | 73.1 | 73.2 | 73.4 | 72.9 | ||
9 | KM598415.1 USA | 94.2 | 94.3 | 94.3 | 94.3 | 94.9 | 94.9 | 93.6 | 95.5 | - | 90.9 | 91.9 | 73.6 | 73.6 | 73.8 | 73.2 | ||
10 | KM598416.1 USA | 96.7 | 96.8 | 96.8 | 96.8 | 97.7 | 97 | 96.2 | 96.7 | 95.7 | - | 88.4 | 73.6 | 73.4 | 73.7 | 73.1 | ||
11 | NC_024452.1ABU-P1 HUN | 92.8 | 93 | 93 | 93 | 93 | 93.3 | 92.7 | 94.8 | 96.8 | 93 | - | 73.4 | 73.1 | 73.4 | 72.8 | ||
TuPV | 12 | KM598418.1 USA | 79.6 | 79.7 | 79.7 | 79.7 | 79.7 | 79.3 | 78.7 | 78.7 | 79.4 | 79.4 | 78.8 | - | 98.3 | 98.8 | 98.1 | |
13 | KM598419.1 USA | 78.8 | 79 | 79 | 79 | 79 | 78.5 | 78.1 | 78.2 | 78.8 | 78.7 | 78.2 | 98.2 | - | 99.5 | 98.8 | ||
14 | NC_024454.1 1078 USA | 79.6 | 79.7 | 79.7 | 79.7 | 79.7 | 79.3 | 78.8 | 79 | 79.6 | 79.4 | 79 | 99.4 | 98.8 | - | 99.3 | ||
15 | KM598420.1 USA | 77.8 | 77.9 | 77.9 | 77.9 | 77.9 | 77.5 | 77.1 | 77.2 | 77.9 | 77.6 | 77.4 | 97.6 | 97 | 98.2 | - | ||
Amino acids (%) |
Age of Post-Infection in Days | Clinical Signs | Postmortem Examination | ||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Depression | Ruffles Feather | Apathy | Somnolence | Lethargy | Runting | Stunting | Duodenum Loop Enrolled | Foamy Content | Aqueous Feces | Undigested Feed | Intestinal Segmentation | Pancreas Atrophy | Yolk Sac Un-Absorbed | |||||||||
D | J | I | D | J | I | D | J | I | D | J | I | |||||||||||
0 | − (5/5) | − (5/5) | − (5/5) | − (5/5) | - (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) | − (5/5) |
1 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | − (5/5) | − (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
2 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | − (5/5) | − (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
3 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | − (5/5) | − (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
4 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (4/5) | + (4/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
5 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
6 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
7 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
14 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (4/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) |
21 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (4/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (4/5) | + (5/5) |
28 | + (4/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (4/5) | + (4/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (4/5) | + (4/5) |
35 | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (3/5) | + (3/5) | + (4/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (4/5) | + (5/5) |
42 | + (3/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (3/5) | + (3/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (5/5) | + (3/5) |
Organ | Day of Harvest | Group | Mesenteritis | Crypt Morphology | Lamina Propria | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Crypt Hyperplasia | Cyst-Shape | Necrosis | Number of Cells in Mitosis | p-Value | Follicles | Heterophil | Lymphocytes | Intraepithelial Lymphocytes | p-Value | ||||||
Duodenum | 7 Days | Mock | 6.5 | ±1.19 | * 0.05 | 4.7 | ±0.88 | * 0.03125 | |||||||
ChPV | + (5/5) | + (2/5); ++ (3/5) * | + (3/5) | + (5/5) | 12 | ±6.09 | + (1/5) | 11.5 | 1.86 | ||||||
14 Days | Mock | 5.1 | ±0.92 | 2.1 | ±0.37 | * 0.03125 | |||||||||
ChPV | + (4/5) | + (2/5); ++ (3/5) * | + (3/5) | + (5/5) | 6.4 | ±2.03 | + (1/5) | 11.6 | ±3.72 | ||||||
21 Days | Mock | 3.4 | ±0.43 | * 0.03125 | 6.8 | ±1.3 | * 0.02895 | ||||||||
ChPV | + (4/5) | + (2/5); ++ (3/5) * | 11.1 | ±3.6 | + (2/5) | + (2/5) | 16.7 | ±1.15 | |||||||
28 Days | Mock | 4.8 | ±1.12 | * 0.03125 | 6.1 | ±1.84 | |||||||||
ChPV | + (5/5) | + (3/5) *; ++ (2/5) | + (3/5) | 15.9 | ±4.64 | + (1/5) | + (3/5) * | 15.6 | ±1.10 | ||||||
35 Days | Mock | 4 | ±1.22 | * 0.03125 | 6.1 | ±2.23 | |||||||||
ChPV | + (5/5) | + (5/5) * | 10.5 | ±2.06 | + (1/5) | + (1/5) | 13 | ±2.08 | |||||||
42 Days | Mock | 5.6 | ±0.79 | * 0.02895 | + (1/5) | 5.1 | ±0.69 | ||||||||
ChPV | + (3/5) | + (3/5) *; ++ (2/5) | + (3/5) | 17.9 | ±3.39 | + (1/5) | + (1/5) | 14 | ±1.25 | ||||||
Jejunum | 7 Days | Mock | 4.4 | ±1.26 | * 0.03125 | 5.9 | ±0.88 | ||||||||
ChPV | + (4/5) *; ++ (1/5) | + (2/5) | + (4/5) | 9.3 | ±4.38 | 7.9 | ±4.43 | ||||||||
14 Days | Mock | 5.5 | ±1.02 | 3.7 | ±1.09 | * 0.03125 | |||||||||
ChPV | + (1/5); ++ (4/5) * | + (3/5) | 8.2 | ±2.63 | + (3/5) * | 10.3 | ±3.21 | ||||||||
21 Days | Mock | + (5/5) | 4.9 | ±1.12 | * 0.03125 | 10.6 | ±0.69 | * 0.03126 | |||||||
ChPV | + (5/5) * | + (5/5) | 10.2 | ±2.93 | + (1/5) | + (1/5) | 16.3 | ±4.38 | |||||||
28 Days | Mock | 8.1 | ±1.36 | 9.9 | ±2.63 | ||||||||||
ChPV | + (1/5); ++ (4/5) * | + (1/5) | 15.5 | ±5.69 | + (1/5) | + (1/5); ++ (1/5) | 15 | ±5.69 | |||||||
35 Days | Mock | 8.3 | ±1.96 | 11.1 | ±3.67 | ||||||||||
ChPV | + (2/5) | + (3/5) *; ++ (2/5) | 10.2 | ±4.02 | + (1/5) | + (5/5) * | 11.8 | ±2.38 | |||||||
42 Days | Mock | 6.1 | ±1.48 | 4 | ±0.88 | ||||||||||
ChPV | + (2/5); ++ (2/5) | + (2/5) | 9.5 | ±2.56 | + (2/5) | 11.8 | ±2.33 | ||||||||
Ileum | 7 Days | Mock | 3.6 | ±0.38 | * 0.05 | 4.9 | ±1.54 | ||||||||
ChPV | + (3/5) *; ++ (2/5) | + (3/5) | + (4/5) | 7.7 | ±2.81 | 9.2 | ±2.96 | ||||||||
14 Days | Mock | 4.6 | ±0.69 | * 0.03125 | 4.9 | ±0.57 | |||||||||
ChPV | + (2/5); ++ (3/5) * | + (2/5) | 11.7 | ±2.92 | 15.3 | ±2.96 | |||||||||
21 Days | Mock | 4 | ±1.19 | * 0.03125 | 3.7 | ±0.33 | * 0.04876 | ||||||||
ChPV | + (3/5) *; ++ (2/5) | + (3/5) | + (5/5) | 7.5 | ±2.55 | 11.3 | ±1.57 | ||||||||
28 Days | Mock | 5.1 | ±0.73 | 3.9 | ±1.71 | * 0.03125 | |||||||||
ChPV | + (5/5) * | + (5/5) | 12.2 | ±4.92 | 10.9 | ±1.8 | |||||||||
35 Days | Mock | 5.7 | ±0.89 | * 0.03125 | 4 | ±0.27 | * 0.03125 | ||||||||
ChPV | + (3/5) *; ++ (2/5) | 13 | ±3.41 | + (4/5) *; ++ (1/5) | 13.8 | ±0.38 | |||||||||
42 Days | Mock | 4.9 | ±0.51 | * 0.03125 | + (2/5) | 3.6 | ±0.5 | ||||||||
ChPV | + (5/5) * | 14.3 | ±4.86 | + (1/5) | + (3/5) *; ++ (1/5) | 12.1 | ±2.5 |
Tissues/Organs * | Detection and Quantification of ChPV in the Tissues through qPCR at Different Days Post Infection | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 14 | 21 | 28 | 35 | 42 | |
Duodenum | −/0 | +/31 ** | +/81 | +/1085 | +/1612 | +/583 | +/1170 | +/9863 | +/116054 | +/24197 | +/4921 | +/159 | +/50 |
Jejunum | −/0 | +/884 | +/911 | +/665 | +/96 | +/665 | +/1554 | +/85808 | +/122531 | +/15258 | +/3100 | +/844 | +/146 |
Ileum | −/0 | +/1676 | +/1468 | +/14441 | +/19261 | +/28822 | +/21594 | +/26644 | +/206642 | +/121564 | +/1470 | +/6855 | +/97 |
Spleen | −/0 | + | + | + | + | + | +/4606 | +/1570 | +/8002 | +/31198 | +/8364 | +/7365 | +/1027 |
Thymus | −/0 | +/111 | +/50 | +/1140 | +/3452 | +/1267 | +/1610 | +/56 | +/82 | +/323 | +/688 | +/788 | +/1767 |
Liver | −/0 | +/54 | +/60 | +/667 | +/8163 | +/6430 | +/3899 | +/832 | +/5898 | +/7598 | +/3684 | +/3294 | +/1473 |
Kidney | −/0 | +/221 | +/33 | +/140 | +/936 | +/1229 | +/618 | +/101 | +/521 | +/91 | +/473 | +/44 | +/667 |
Pancreas | −/0 | +/59 | +/50 | +/62 | +/87 | +/257 | +/210 | +/70 | +/105 | +/230 | +/371 | +/1078 | +/1250 |
Proventriculus | −/0 | +/11 | +/21 | +/65 | +/1374 | +/426 | +/3124 | +/3527 | +/4850 | +/517 | +/489 | +/241 | +/295 |
Bursa | −/0 | + | + | + | + | +/592 | +/9230 | +/6000 | +/5590 | +/3979 | +/2345 | +/2300 | +/2483 |
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N. Nuñez, L.F.; Santander-Parra, S.H.; De la Torre, D.I.; Sá, L.R.M.d.; Buim, M.R.; Astolfi-Ferreira, C.S.; Piantino Ferreira, A.J. Molecular Characterization and Pathogenicity of Chicken Parvovirus (ChPV) in Specific Pathogen-Free Chicks Infected Experimentally. Pathogens 2020, 9, 606. https://doi.org/10.3390/pathogens9080606
N. Nuñez LF, Santander-Parra SH, De la Torre DI, Sá LRMd, Buim MR, Astolfi-Ferreira CS, Piantino Ferreira AJ. Molecular Characterization and Pathogenicity of Chicken Parvovirus (ChPV) in Specific Pathogen-Free Chicks Infected Experimentally. Pathogens. 2020; 9(8):606. https://doi.org/10.3390/pathogens9080606
Chicago/Turabian StyleN. Nuñez, Luis Fabian, Silvana H. Santander-Parra, David I. De la Torre, Lilian R. M. de Sá, Marcos R. Buim, Claudete S. Astolfi-Ferreira, and Antonio J. Piantino Ferreira. 2020. "Molecular Characterization and Pathogenicity of Chicken Parvovirus (ChPV) in Specific Pathogen-Free Chicks Infected Experimentally" Pathogens 9, no. 8: 606. https://doi.org/10.3390/pathogens9080606