Abstract: Strangles is a highly contagious disease of the equine upper respiratory tract caused by Streptococcus equi subspecies. Streptococcus equi subsp. equi (S. equi) and Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) was isolated, as local, hot, and field strains, from horses clinically suffering from respiratory distress. The isolated Streptococci were identified using bacteriological and molecular techniques. Four formulations of inactivated S. equi vaccines were developed and evaluated. The first formulation was prepared using the S. equi isolates, adjuvanted with MONTANIDE GEL adjuvant, while the second formulation was adjuvanted with MONTANIDE ISA-70 adjuvant. The other 2 formulations were inactivated combined vaccines prepared from both S. equi and S. zooepidemicus isolates. The 3rd formulation was the combined isolates adjuvanted with MONTANIDE GEL while the 4th formulation was the combined isolates adjuvanted with MONTANIDE ISA-70. The developed vaccines' physical properties, purity, sterility, safety, and potency were ensured. The immunizing efficacy was determined in isogenic BALB/c mice and white New Zealand rabbits using the passive hemagglutination test. Also, the antibodies' titer of the combined S. equi and S. zooepidemicus vaccine adjuvanted with MONTANIDE ISA-70 in foals was tracked using an indirect enzyme-linked immunosorbent assay. The protective efficacy of the developed vaccines was determined using a challenge test in both laboratory and field animal models, where a 75% protection rate was achieved. The combined vaccine proved to be more efficacious than the monovalent vaccine. Also, the MONTANIDE ISA-70 adjuvant provided significant protective efficacy than the MONTANIDE GEL. The current work is introducing a very promising mitigative and strategic controlling solution for strangles.
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Overview
This research article focuses on developing new vaccines against strangles, a contagious respiratory disease in horses caused by Streptococcus equi subspecies.
It evaluates four different inactivated vaccine formulations for their immunizing potential and protective efficacy using laboratory animals and foals.
Background on Strangles and Streptococcus equi
Strangles is a common, highly contagious infection affecting the upper respiratory tract of horses.
The disease is caused primarily by Streptococcus equi subsp. equi (S. equi), but Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) can also be involved.
The researchers isolated these bacteria strains from horses showing clinical signs of respiratory distress during outbreaks.
Both bacteriological culture and molecular methods were used to accurately identify the isolated strains.
Vaccine Development
Four different inactivated vaccine formulations were created:
Formulation 1: Inactivated S. equi isolates adjuvanted with MONTANIDE GEL.
Formulation 2: Inactivated S. equi isolates adjuvanted with MONTANIDE ISA-70.
Formulation 3: Combined inactivated S. equi and S. zooepidemicus isolates adjuvanted with MONTANIDE GEL.
Formulation 4: Combined inactivated S. equi and S. zooepidemicus isolates adjuvanted with MONTANIDE ISA-70.
The adjuvants, MONTANIDE GEL and MONTANIDE ISA-70, are used to enhance immune responses triggered by the vaccines.
Quality Assurance of Vaccines
The physical aspects of the vaccines such as appearance and consistency were confirmed to meet standards.
Rigorous purity and sterility tests ensured no contamination was present in the vaccine preparations.
Safety tests were conducted to confirm that the vaccines did not induce adverse effects in administered animals.
Potency tests measured the ability of the vaccine to induce an adequate immune response.
Assessment of Immunogenicity
The vaccines were tested in two laboratory animal models: isogenic BALB/c mice and white New Zealand rabbits.
Immunizing efficacy was assessed using the passive hemagglutination test, which detects antibody levels against Streptococcus strains.
For foals, the immune response to the combined vaccine containing both S. equi and S. zooepidemicus with MONTANIDE ISA-70 was tracked over time using an indirect enzyme-linked immunosorbent assay (ELISA), a sensitive method for antibody detection.
Evaluation of Protective Efficacy
The vaccines’ protective abilities were tested via challenge experiments, where animals were exposed to virulent Streptococcus strains after vaccination.
Both laboratory and field models were employed to simulate real-world conditions.
Results showed up to 75% protection in vaccinated animals, indicating substantial vaccine efficacy.
The combined vaccine (S. equi + S. zooepidemicus) demonstrated better protection than the monovalent vaccine (S. equi alone).
The formulation using MONTANIDE ISA-70 adjuvant resulted in significantly higher protection compared to the MONTANIDE GEL adjuvant.
Conclusion and Implications
This study presents promising new vaccine candidates to control and mitigate strangles in horses.
By combining two subspecies of Streptococcus equi and optimizing the choice of adjuvant, improved vaccine efficacy was achieved.
The findings suggest that the combined vaccines, particularly with MONTANIDE ISA-70, could form an effective strategic approach for managing strangles outbreaks in equine populations.
Further field studies and eventual commercialization could lead to better preventive measures to reduce the incidence and impact of this economically significant equine disease.
Cite This Article
APA
Soliman R, Yousef M, Gelil SA, Aboul-Ella H.
(2024).
Development of novel Streptococcus equi vaccines with an assessment of their immunizing potentials and protective efficacies.
BMC Vet Res, 20(1), 173.
https://doi.org/10.1186/s12917-024-04012-z
Ivens PA, Matthews D, Webb K, Newton JR, Steward K, Waller AS, Robinson C, Slater JD. Molecular characterization of ‘strangles’ outbreaks in the UK: the use of M-protein typing of Streptococcus equi ssp. equi.. Equine Vet J 2011;43(3):359–64.
Webb K, Barker C, Harrison T, Heather Z, Steward KF, Robinson C, Newton JR, Waller AS. Detection of Streptococcus equi subspecies equi using a triplex qPCR assay.. Vet J 2013;195(3):300–4.
Arafa AA, Hedia RH, Ata NS, Ibrahim ES. Vancomycin-resistant Streptococcus equi subsp. equi isolated from equines suffering from respiratory manifestation in Egypt.. Vet World 2021;14(7):1808–14.
Hemeda SM, Sayed RH, Hassan H, Sheima AE, Aboul-Ella H, Soliman R. Development of colloid gold-based lateral flow immunochromatographic kits for screening and rapid detection of beta-lactams antibiotic residues in dairy milk.. Adv Anim Vet Sci 2022;10(7):1616–22.
Hashem Z, Soliman R, Aziz MAE, Badr Y, Aboul-Ella H. Development, preparation, and evaluation of a murine monoclonal antibodies-based interferon-gamma enzyme-linked immunosorbent assay (ELISA) for rapid and accurate diagnosis of bovine tuberculosis.. Int J Veterinary Sci Int J Vet Sci 2023;12(4):599–606.
Sally R, Rafik S, Manal AA, Lamiaa O, Ahmed S, Heidy A, Hassan A. Preparation of a newly developed trivalent Pasteurella multocida, Avibacterium paragallinarum, and Ornithobacterium rhinotrachealeVaccine with an evaluation of its protective efficacy in chickens.. J Adv Vet Res 2023;13(5):737–42.
Soliman R, Waheeb N, Nasr E, El-Hariri M, Abo-Elyazeed H, Aboul-Ella H. Critical field appraisal for the use of bovine tuberculosis’ antibody detecting-serodiagnostics.. Adv Anim Vet Sci 2023;11(2):278–87.
Soliman R, Abd Elbaki S, Youssef M, Abdelmalak G, Aboul-Ella H. Production of Rhodococcus equi-specific horse polyclonal antibodies and evaluation of their therapeutic efficacy and prophylactic potentials in a mouse model.. Bulg J Vet Med 2024;online first.
Soliman R, Hashem Z, Abo-Elyazeed H, Aboul-Ella H. Development and production of Rabies virus-specific monoclonal antibodies and evaluation of their neutralizing and protective potentials.. Microbes Infect Dis 2024.
Sawant AA, Pillai SR, Jayarao BM. Evaluation of five selective media for isolation of catalase-negative gram-positive cocci from bulk tank milk.. J Dairy Sci 2002;85(5):1127–32.
Timoney JF, Artiushin SC, Boschwitz JS. Comparison of the sequences and functions of Streptococcus equi M-like proteins SeM and SzPSe.. Infect Immun 1997;65(9):3600–5.
. Animals and Animal products. Standard requirements, part 1: Sects. 113, 1996, Washington, USA Government printing office.. Code Federal Regulation 9. CFR9 1550.
Bazid AH, Amer HM, Nayel M, Attia M, Maklad N, Wasfy M, Abdelmegeid M, El-Sayed MM, Magouz A, Badr Y. Assessment of the potency and effectiveness of a heptavalent oil-adjuvanted (ISA 206) foot-and-mouth disease vaccine in Egypt.. Arch Virol 2023;168(2):62.
Yi L, Wang Y, Ma Z, Lin HX, Xu B, Grenier D, Fan HJ, Lu CP. Identification and characterization of a Streptococcus equi ssp. zooepidemicus immunogenic GroEL protein involved in biofilm formation.. Vet Res 2016;47:50.
Knowles EJ, Mair TS, Butcher N, Waller AS, Wood JL. Use of a novel serological test for exposure to Streptococcus equi subspecies equi in hospitalized horses.. Vet Rec 2010;166(10):294–7.
Moraes CM, Conceição FR, Rocha ASR, Santos Júnior AG, Ribas LM, Vagas APC, Nogueira CEW, Gil-Turnes C, Leite FBL. Cloning, expression, and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA.. Arq Bras Med Vet Zootec 2014;664:1015–22.
Jacobs JA, Schot CS, Schouls LM. The Streptococcus anginosus species comprises five 16S rRNA ribogroups with different phenotypic characteristics and clinical relevance.. Int J Syst Evol Microbiol 2000;3:1073–9.
Aucouturier J, Dupuis L, Deville S, Ascarateil S, Ganne V. Montanide ISA 720 and 51: a new generation of water in oil emulsions as adjuvants for human vaccines.. Expert Rev Vaccines 2002;1:111–8.
Facciolà A, Visalli G, Laganà A, Di Pietro A. An overview of Vaccine adjuvants: current evidence and future perspectives.. Vaccines (Basel) 2022;10(5):819.
Galán JE, Timoney JF. Immunologic and genetic comparison of Streptococcus equi isolates from the United States and Europe. J Clin Microbiol 1988;26(6):1142–6.
Fahmy BGA, El-Genaidy HM. Biochemical and pathological profilesin streptococci infection in imported horses. Egypt J Comp Pathol Clin Pathol 2010;23(1).
Galán JE, Timoney JF. Molecular analysis of the M protein of Streptococcus equi and cloning and expression of the M protein gene in Escherichia coli. Infect Immun 1987;55(12):3181–7.
Meehan M, Muldowney DA, Watkins NJ, Owen P. Localization and characterization of the ligand-binding domain of the fibrinogen-binding protein (FgBP) of Streptococcus equi subsp. equi. Microbiology 2000;146(5):1187–94.