Evaluation of Contamination Risks with Coxsackievirus B4 E2 in Swiss Albino Mice Stools
Coxsackie B4 (CV-B4), is a major cause of viral myocarditis, dilated cardiomyopathy, and pancreatitis. Like other human enteroviruses, CV-B4 is ubiquitous, excreted in the stool, transmitted by fecal–oral route, and persists in the environment. In the context of studies on CV-B4 infection, it is important to investigate how this virus can be eliminated and to show the possibility of contamination risk with a CV-B4 E2 infected Swiss albino mice. Swiss albino female mice were inoculated with CV-B4 E2 strain and divided in two groups: the first was intraperitoneally (I.P.) infected; the second was orally infected. In order to study the CV-B4 E2 infection in mice, total RNA was extracted from thymus, spleen, pancreas, and intestine, and viral genome was detected using semi-nested (RT-PCR). To further demonstrate infection or immunization of mice, Sera obtained from infected mice were assayed in vitro for their neutralizing antibody. To detect virus in stool of infected mice, stool samples were collected at different post-infection (p.i.) times. Neutralizing antibodies were detectable all along the follow-up period (Day 0, 1, 3, 7, 9, 17, 22, 30, 45, 60 p.i.) in I.P and oral infected mice. Our results showed that when mice were inoculated successively at day 0 and day 8, neutralizing activity was increased in I.P route more than in the oral route. Viral isolation in HEp-2 cells showed negative results. Stool viral analyses reveal a low detection of the CV-B4 E2 genome for all infected mice. In conclusion, our experiments demonstrated that there are no risks linked with the stool of CV-B4 E2 of Swiss albino mice. It would be interesting to characterize the inhibitors of the virus infectivity in these biological samples (stool) and investigate their targets and mechanisms of action.
Leila Aguech-Oueslati was supported by the Comité Mixte de Coopération Universitaire Franco-Tunisian (CMCU 08/G808) with grants from Egide Paris. This work was supported by the Ministry for Scientific Research, Technology and the Development of Competencies, (LR99ES27), Tunisia, Ministry for the National Education of Research and Technology, Université de Lille, et CHU de Lille Laboratoire de Virologie EA3610, F-59037, Lille, France.
- 3.Bopegamage S, Borsanyiova M, Vargova A, Petrovicova A, Benkovicova M, Gomolcak P (2003) Coxsackievirus infection of mice I. Viral kinetics and histopathological changes in mice experimentally infected with coxscakievirus B3 and B4 by the oral route. Act Virol 47:245–251Google Scholar
- 4.Bopegamage S, Kovacova J, Vargova A, Motusova J, Petrovicova A, Benkovicova M, Gomolcak P, Bakkers J, van Kuppeveld F, Melchers WJG, Galama JM (2005) Coxsackie B virus infection of mice: inoculation by the oral route protects the pancreas from damage, but not from infection. J Gen Virol 86:3271–3280CrossRefPubMedGoogle Scholar
- 8.Elmastour F, Jaidane H, Aguech-Oueslati L, Benkahla MA, Aouni M, Gharbi J, Sane F, Hober D (2016) Immunoglobulin G-dependent enhancement of the infection with coxsackievirus B4 in a murine system. Virulence 30:1–9Google Scholar
- 9.Elmastour F, Jaïdane H, Ben Kahla MA, Aguech-Oueslati L, Sané F, Aymen Halouani A, Engelmann I, Bertin A, Mokni M, Gharbi J, Aouni M, Alidjinou EK, Hober D (2016) Anti-coxsackievirus B4 (CV-B4) enhancing activity of serum associated with increased viral load and pathology in mice reinfected with CV-B4. Virulence. doi: 10.1080/21505594.2016.1252018 Google Scholar
- 17.Knowles NJ, Hovi T, Hyypiä T, King AMQ, Lindberg M, Pallansch MA, Palmenberg AC, Simmonds P, Skern T, Stanway G (2012) Picornaviridae. In: King AMQ, Adams MJ, Carstens EB, LefkowitzEJ (eds) In virus taxonomy: classification and nomenclature of viruses: Ninth Report of the International Committee on taxonomy of viruses. Elsevier, San Diego, CA, pp 855–880Google Scholar
- 18.Leparc I, Aymard M, Acute Fuchs F (1994) Chronic and persistent enterovirus and poliovirus infections: detection of viral genome by seminested PCR amplification in culture-negative samples. Mol Cell 8:487–495Google Scholar
- 20.Melnick JL (1996) Enteroviruses: polioviruses, coxsackieviruses, echoviruses, and newer enteroviruses. In: Fields BN, Knipe DM (eds) Virology. Raven Press, New York, pp 549–605Google Scholar
- 25.Retoo KN, Osman SA, Illavia SJ, Cameron-Wilson CL, Banatvala JE, Muir P (2001) Quantitative analysis of viral RNA kinetic in coxsackievirus B3-induced murine myocarditis: biphasic pattern of clearance following acute infection with persistence of residual viral RNA throughout and beyond the inflammatory phase of disease. J Gen Virol 81(Pt11):2755–2762Google Scholar
- 26.Reed LJ, Muench H (1938) A simple method of estimating fifty percent end points. Am J Hyg 27:493–497Google Scholar