Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in Türkiye

Abstract Background Clostridial diseases are common in ruminants all over the world. This study investigated the efficacy of a novel recombinant vaccine developed against enterotoxemia and infectious necrotic hepatitis in sheep. Results Plasmids carrying the Clostridium novyi type B alpha toxin gene...

Full description

Saved in:
Bibliographic Details
Main Authors: Dilek Nur Ekinoğlu, Venhar Çelik, Esra Gül, Hakan Kalender, Hasan Öngör, Burhan Çetinkaya
Format: Article
Language:English
Published: BMC 2025-05-01
Series:BMC Veterinary Research
Subjects:
Online Access:https://doi.org/10.1186/s12917-025-04841-6
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849704173225902080
author Dilek Nur Ekinoğlu
Venhar Çelik
Esra Gül
Hakan Kalender
Hasan Öngör
Burhan Çetinkaya
author_facet Dilek Nur Ekinoğlu
Venhar Çelik
Esra Gül
Hakan Kalender
Hasan Öngör
Burhan Çetinkaya
author_sort Dilek Nur Ekinoğlu
collection DOAJ
description Abstract Background Clostridial diseases are common in ruminants all over the world. This study investigated the efficacy of a novel recombinant vaccine developed against enterotoxemia and infectious necrotic hepatitis in sheep. Results Plasmids carrying the Clostridium novyi type B alpha toxin gene (CnBtcn-alpha), Clostridium perfringens type C beta toxin gene (CpCcpb) and C. perfringens type D epsilon toxin gene (CpDetx) were constructed and the plasmids were transferred to Escherichia coli. Unpurified protein obtained from E. coli cell lysate supernatant was used to prepare the recombinant vaccine. The vaccine was prepared in three different formulas (RV100, RV200 and RV400), with 100, 200 and 400 µg of each protein in one vaccine dose. RV400 was preferred to immunise sheep as the antitoxin titres in the pooled blood serum of rabbits administered with this vaccine were determined to be above the minimum values specified in European Pharmacopoeia (10 IU/mL for beta, 5 IU/mL for epsilon, and 3.5 IU/mL for alpha). A total of 24 Akkaraman breed sheep with no antibodies against the relevant toxins were used for the experiment. All the animals in three groups (recombinant vaccine group, commercial polyvalent vaccine group and negative control group), each consisting of eight sheep, were vaccinated twice with an interval of 21 days and, the antitoxin titres were measured 14 days after the second vaccination by the mouse toxin neutralization test. The average antitoxin titres in sheep immunised with RV400 were calculated as 9.75 ± 1.28 IU/mL for C. perfringens beta, 13.75 ± 1.98 IU/mL for C. perfringens epsilon and 5.50 ± 0.93 IU/mL for C. novyi alpha toxins. On the other hand, the average values in sheep immunised with commercial vaccine were detected as 8.00 ± 2.14 IU/mL, 4.25 ± 1.67 IU/m L and 6.50 ± 0.93 IU/mL for C. perfringens beta, C. perfringens epsilon and C. novyi alpha, respectively. No antitoxin titre was detected in sheep in the negative control group (PBS). A statistically significant difference was observed between the recombinant and commercial vaccine groups in terms of C. perfringens epsilon antitoxin titres (P = 0.0002). Conclusions The present study was the first to investigate the efficacy of a combined recombinant vaccine prepared from unpurified proteins against enterotoxemia and infectious necrotic hepatitis and, the results suggested that it shows promise for protecting sheep against these diseases.
format Article
id doaj-art-52aae8e8edf7476e8658dca24fb33cb5
institution DOAJ
issn 1746-6148
language English
publishDate 2025-05-01
publisher BMC
record_format Article
series BMC Veterinary Research
spelling doaj-art-52aae8e8edf7476e8658dca24fb33cb52025-08-20T03:16:51ZengBMCBMC Veterinary Research1746-61482025-05-0121111110.1186/s12917-025-04841-6Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in TürkiyeDilek Nur Ekinoğlu0Venhar Çelik1Esra Gül2Hakan Kalender3Hasan Öngör4Burhan Çetinkaya5Graduate School of Natural and Applied Sciences, Firat UniversityFaculty of Engineering, Department of Bioengineering, Firat UniversityGraduate School of Natural and Applied Sciences, Firat UniversityFaculty of Veterinary Medicine, Department of Microbiology, Firat UniversityFaculty of Veterinary Medicine, Department of Microbiology, Firat UniversityFaculty of Veterinary Medicine, Department of Microbiology, Firat UniversityAbstract Background Clostridial diseases are common in ruminants all over the world. This study investigated the efficacy of a novel recombinant vaccine developed against enterotoxemia and infectious necrotic hepatitis in sheep. Results Plasmids carrying the Clostridium novyi type B alpha toxin gene (CnBtcn-alpha), Clostridium perfringens type C beta toxin gene (CpCcpb) and C. perfringens type D epsilon toxin gene (CpDetx) were constructed and the plasmids were transferred to Escherichia coli. Unpurified protein obtained from E. coli cell lysate supernatant was used to prepare the recombinant vaccine. The vaccine was prepared in three different formulas (RV100, RV200 and RV400), with 100, 200 and 400 µg of each protein in one vaccine dose. RV400 was preferred to immunise sheep as the antitoxin titres in the pooled blood serum of rabbits administered with this vaccine were determined to be above the minimum values specified in European Pharmacopoeia (10 IU/mL for beta, 5 IU/mL for epsilon, and 3.5 IU/mL for alpha). A total of 24 Akkaraman breed sheep with no antibodies against the relevant toxins were used for the experiment. All the animals in three groups (recombinant vaccine group, commercial polyvalent vaccine group and negative control group), each consisting of eight sheep, were vaccinated twice with an interval of 21 days and, the antitoxin titres were measured 14 days after the second vaccination by the mouse toxin neutralization test. The average antitoxin titres in sheep immunised with RV400 were calculated as 9.75 ± 1.28 IU/mL for C. perfringens beta, 13.75 ± 1.98 IU/mL for C. perfringens epsilon and 5.50 ± 0.93 IU/mL for C. novyi alpha toxins. On the other hand, the average values in sheep immunised with commercial vaccine were detected as 8.00 ± 2.14 IU/mL, 4.25 ± 1.67 IU/m L and 6.50 ± 0.93 IU/mL for C. perfringens beta, C. perfringens epsilon and C. novyi alpha, respectively. No antitoxin titre was detected in sheep in the negative control group (PBS). A statistically significant difference was observed between the recombinant and commercial vaccine groups in terms of C. perfringens epsilon antitoxin titres (P = 0.0002). Conclusions The present study was the first to investigate the efficacy of a combined recombinant vaccine prepared from unpurified proteins against enterotoxemia and infectious necrotic hepatitis and, the results suggested that it shows promise for protecting sheep against these diseases.https://doi.org/10.1186/s12917-025-04841-6EnterotoxemiaInfectious necrotic hepatitisImmune responseRecombinant vaccineSheep
spellingShingle Dilek Nur Ekinoğlu
Venhar Çelik
Esra Gül
Hakan Kalender
Hasan Öngör
Burhan Çetinkaya
Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in Türkiye
BMC Veterinary Research
Enterotoxemia
Infectious necrotic hepatitis
Immune response
Recombinant vaccine
Sheep
title Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in Türkiye
title_full Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in Türkiye
title_fullStr Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in Türkiye
title_full_unstemmed Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in Türkiye
title_short Sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in Türkiye
title_sort sheep immune response against a novel recombinant enterotoxemia and infectious necrotic hepatitis vaccine in turkiye
topic Enterotoxemia
Infectious necrotic hepatitis
Immune response
Recombinant vaccine
Sheep
url https://doi.org/10.1186/s12917-025-04841-6
work_keys_str_mv AT dileknurekinoglu sheepimmuneresponseagainstanovelrecombinantenterotoxemiaandinfectiousnecrotichepatitisvaccineinturkiye
AT venharcelik sheepimmuneresponseagainstanovelrecombinantenterotoxemiaandinfectiousnecrotichepatitisvaccineinturkiye
AT esragul sheepimmuneresponseagainstanovelrecombinantenterotoxemiaandinfectiousnecrotichepatitisvaccineinturkiye
AT hakankalender sheepimmuneresponseagainstanovelrecombinantenterotoxemiaandinfectiousnecrotichepatitisvaccineinturkiye
AT hasanongor sheepimmuneresponseagainstanovelrecombinantenterotoxemiaandinfectiousnecrotichepatitisvaccineinturkiye
AT burhancetinkaya sheepimmuneresponseagainstanovelrecombinantenterotoxemiaandinfectiousnecrotichepatitisvaccineinturkiye