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Design, synthesis, and evaluation of peptides derived from L1 protein against bovine papillomavirus-1/2 identified along Mexico’s cattle export route

Paola Leonor García Coronado
Paola Leonor García Coronado
, 
César Iván Romo Sáenz
César Iván Romo Sáenz
, 
Jorge R. Kawas
Jorge R. Kawas
, 
Diana Ginette Zarate Triviño
Diana Ginette Zarate Triviño
, 
Yareellys Ramos Zayas
Yareellys Ramos Zayas
, 
Silvia Elena Santana Krímskaya
Silvia Elena Santana Krímskaya
, 
Cristina Rodríguez Padilla
Cristina Rodríguez Padilla
 and 
Moisés Armides Franco Molina
Moisés Armides Franco Molina
   | Feb 28, 2023
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Published Online: Feb 28, 2023
Page range: 11 - 21
Received: May 23, 2022
Accepted: Jan 26, 2023
DOI: https://doi.org/10.2478/jvetres-2023-0003
Keywords
© 2023 P.L. García Coronado et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

Bovine papillomavirus BPV-1 and BPV-2 shown in representative samples which were positive for viral genetic material when electrophoresed in 1.5% agarose gel M – 100 base pair molecular weight marker; C+ – BPV-1 positive sample from Nuevo León; C− – DNA-free reaction mix; A – BPV-1 presence in cutaneous/genital warts: lanes 1–11, 13–14 and 16–25; B – BPV-2 presence in cutaneous/genital warts: lanes 8–11, 13–14 and 18–25 (Nuevo León, lanes 1–5 and 24–25; Veracruz, 18–23; Chiapas, 6–7 and 14–17; and Tabasco, 8–13)
Bovine papillomavirus BPV-1 and BPV-2 shown in representative samples which were positive for viral genetic material when electrophoresed in 1.5% agarose gel M – 100 base pair molecular weight marker; C+ – BPV-1 positive sample from Nuevo León; C− – DNA-free reaction mix; A – BPV-1 presence in cutaneous/genital warts: lanes 1–11, 13–14 and 16–25; B – BPV-2 presence in cutaneous/genital warts: lanes 8–11, 13–14 and 18–25 (Nuevo León, lanes 1–5 and 24–25; Veracruz, 18–23; Chiapas, 6–7 and 14–17; and Tabasco, 8–13)

Fig. 2

Samples negative for bovine papillomavirus (BPV)-1/2/4 and therefore with genotype unknown A – sample from Emiliano Zapata, Tabasco; B – sample from Libertad, Chiapas. Samples had cauliflower morphology suggesting that they be considered BPV positive
Samples negative for bovine papillomavirus (BPV)-1/2/4 and therefore with genotype unknown A – sample from Emiliano Zapata, Tabasco; B – sample from Libertad, Chiapas. Samples had cauliflower morphology suggesting that they be considered BPV positive

Fig. 3

Bovine papillomavirus (BPV)-1 and BPV-2 genotype distribution per state from representative samples. BPV-1 was found as a unique genotype or as a co-infection with BPV-2, but BPV-2 was found only as a co-infection
Bovine papillomavirus (BPV)-1 and BPV-2 genotype distribution per state from representative samples. BPV-1 was found as a unique genotype or as a co-infection with BPV-2, but BPV-2 was found only as a co-infection

Fig. 4

Bovine papillomavirus (BPV)-1 phylogenetic tree of Mexican isolates (partial L1 sequence). Sequences were compared with sequences from different nationalities recorded in the NCBI database, of which the accession numbers are indicated. The comparison used a neighbour-joining method in MEGA 10.1. RS – representative sample
Bovine papillomavirus (BPV)-1 phylogenetic tree of Mexican isolates (partial L1 sequence). Sequences were compared with sequences from different nationalities recorded in the NCBI database, of which the accession numbers are indicated. The comparison used a neighbour-joining method in MEGA 10.1. RS – representative sample

Fig. 5

Bovine papillomavirus (BPV)-2 phylogenetic tree of Mexican isolates (partial L2 sequence). Sequences were compared with sequences from different nationalities recorded in the NCBI database, of which the accession numbers are indicated. The comparison used a neighbour-joining method in MEGA 10.1. RS – representative sample
Bovine papillomavirus (BPV)-2 phylogenetic tree of Mexican isolates (partial L2 sequence). Sequences were compared with sequences from different nationalities recorded in the NCBI database, of which the accession numbers are indicated. The comparison used a neighbour-joining method in MEGA 10.1. RS – representative sample

Fig. 6

Geographical bovine papillomavirus (BPV) genotype localisation in Mexico. Tabasco, Chiapas, and Veracruz states are considered breeder states and cattle collection centres for Central America. Nuevo León is a final feedlot destination state for national consumption or international exportation. The intensity of grey indicates the percentage of co-infection A – Chiapas; B – Tabasco; C – Veracruz; D – Nuevo León
Geographical bovine papillomavirus (BPV) genotype localisation in Mexico. Tabasco, Chiapas, and Veracruz states are considered breeder states and cattle collection centres for Central America. Nuevo León is a final feedlot destination state for national consumption or international exportation. The intensity of grey indicates the percentage of co-infection A – Chiapas; B – Tabasco; C – Veracruz; D – Nuevo León

Fig. 7

Candidate peptide prediction by online server. The amino acid sequence of the selected peptide possesses antigenicity. Prediction with default values was performed in Bepipred Linear Epitope Predictor
Candidate peptide prediction by online server. The amino acid sequence of the selected peptide possesses antigenicity. Prediction with default values was performed in Bepipred Linear Epitope Predictor

Fig. 8

Specificity of the candidate peptide for BPV-1. BLASTp showed it to be specific to BPV-1 and BPV-2 with 100% homology
Specificity of the candidate peptide for BPV-1. BLASTp showed it to be specific to BPV-1 and BPV-2 with 100% homology

Fig. 9

Sera reactivity against WWL/synthetic peptide and antibody specificity against BPV A – Anti-wart antibody reactivity against WWL at 1: 100 dilution on day 7 and 1:10,000 on days 14 and 35; B – Anti-peptide serum reactivity against synthetic peptide at a titre of 1:1,000,000 on days 7, 14 and 30. Cut-off points (horizontal solid line) were calculated at 0.087 and 0.053, respectively. **** – extremely significant difference (Tukey test P < 0.0001); *** – highly significant difference (Tukey test P < 0.0002) (n = 2); ** – significant difference (Tukey test P < 0.0021); * - significant difference (Tukey test P < 0.0332) (n = 2); C – Anti-wart serum reactivity against peptide at titres of 1 : 10,000–1 : 1,000,000 on day 14 and at a titre of 1 : 10,000 on day 35; D – Anti-peptide serum reactivity against WWL at a titre of 1 : 10,000 on days 7, 14 and 35, and at a titre of 1 : 1,000,000 on days 14 and 35. Letters a, b, and c represent significant difference based on the cut point (horizontal line) of 0.219 in both C and D (Tukey’s test P < 0.05) (n = 2)
Sera reactivity against WWL/synthetic peptide and antibody specificity against BPV A – Anti-wart antibody reactivity against WWL at 1: 100 dilution on day 7 and 1:10,000 on days 14 and 35; B – Anti-peptide serum reactivity against synthetic peptide at a titre of 1:1,000,000 on days 7, 14 and 30. Cut-off points (horizontal solid line) were calculated at 0.087 and 0.053, respectively. **** – extremely significant difference (Tukey test P < 0.0001); *** – highly significant difference (Tukey test P < 0.0002) (n = 2); ** – significant difference (Tukey test P < 0.0021); * - significant difference (Tukey test P < 0.0332) (n = 2); C – Anti-wart serum reactivity against peptide at titres of 1 : 10,000–1 : 1,000,000 on day 14 and at a titre of 1 : 10,000 on day 35; D – Anti-peptide serum reactivity against WWL at a titre of 1 : 10,000 on days 7, 14 and 35, and at a titre of 1 : 1,000,000 on days 14 and 35. Letters a, b, and c represent significant difference based on the cut point (horizontal line) of 0.219 in both C and D (Tukey’s test P < 0.05) (n = 2)

Fig. 10

Anti-wart serum reactivity against WWL at titres of 1 : 100–1 : 10,000 and at a titre of 1 : 100 on day 42 in anti-peptide serum against WWL. The solid line indicates the cut-off point at 0.08; **** – extremely significant difference (Tukey test P < 0.0001); *** – highly significant difference (Tukey test P < 0.0002) (n = 2); ** – significant difference (Tukey test P < 0.0021); * – significant difference (Tukey test P < 0.0332) (n = 2)
Anti-wart serum reactivity against WWL at titres of 1 : 100–1 : 10,000 and at a titre of 1 : 100 on day 42 in anti-peptide serum against WWL. The solid line indicates the cut-off point at 0.08; **** – extremely significant difference (Tukey test P < 0.0001); *** – highly significant difference (Tukey test P < 0.0002) (n = 2); ** – significant difference (Tukey test P < 0.0021); * – significant difference (Tukey test P < 0.0332) (n = 2)

Bovine papillomatosis prevalence in Nuevo León, Chiapas, Veracruz and Tabasco, Mexico

State: Papillomatosis n
Nuevo León
Farm 1 (Linares) 3.77% 450
Farm 2 (Linares) 18% 250
Farm 3 (Cadereyta) 11.43% 700
Chiapas
Farm 4 (Palenque) 12.83% 1200
Farm 5 (Libertad) 11.6% 500
Farm 6 (Salto de Agua) 32.8% 125
Farm 7 (Agua Fria) 20% 50
Veracruz
Farm 8 (Temapache) 10.67% 150
Farm 9 (Tihuatlán) 10% 160
Tabasco
Farm 10 (Emiliano Zapata) 9.2% 1500
Farm 11 (Balancan) 16.25% 80
Farm 12 (Tenosique) 23.44% 320

Amino acid matches between epitope predictions in different software

Online server L1 epitope Position Threshold Score
Bepitope SPATKCASNVIPAK 421 0.75 Yes
ABCpred SILEDTYRYESPATK 410 0.75 0.87
Bepipred PSVLQNWEIGVQPPTSSILEDTYRYIESPATKCASNVIPAKEDPYAGFKF 394 0.75 Yes
LBtope VQPPTSSILEDTYRYIES 404 0.75 85.2
BCEpred DTYRYIESPATKCASNV 414 1.9 Yes

Nucleotide sequence identity analysis of bovine papillomavirus 1 L1 fragments isolated from samples positive for the virus

Sequence Expect (E) value Identity % (alignment with KC595244.2) Gaps
Chiapas (RS7) 7e−129 99% 2/265
Chiapas (RS6) 4e−118 99% 0/242
Nuevo León (RS4) 2e−129 99% 3/267
Nuevo León (RS2) 2e−101 99% 0/209
Nuevo León (RS5) 3e-131 99% 2/266
Veracruz (RS20) 1e−125 99% 1/257
Tabasco (RS8) 1e−125 99% 1/257
Nuevo León (RS3) 2e−126 99% 2/261
Nuevo León (RS1) 6e−128 99% 4/267

Nucleotide sequence identity analysis of bovine papillomavirus 2 L2 fragments isolated from samples positive for the virus

Sequence Expect (E) value Identity % (alignment with MH187961.1) Gaps
Veracruz (RS20) 7e−25 97% 0/71
Tabasco (RS8) 6e−45 91% 3/132
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