MAJOR ARTICLE Ebola Virus Persistence in Body Fluids • OFID • 1 Open Forum Infectious Diseases Received 26 September 2019; editorial decision 26 October 2019; accepted 5 November 2019. a A. K. K. and N. V. contributed equally to this work. Correspondence: Martine Peeters, PhD, IRD, TransVIHMI, 911 avenue Agropolis, 34394 Montpellier Cedex5, France (martine.peeters@ird.fr). Open Forum Infectious Diseases ® © The Author(s) 2019. Published by Oxford University Press on behalf of Infectious Diseases Society of America. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/ by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com DOI: 10.1093/ofd/ofz482 A 40-Month Follow-Up of Ebola Virus Disease Survivors in Guinea (PostEbogui) Reveals Long-Term Detection of Ebola Viral Ribonucleic Acid in Semen and Breast Milk Alpha Kabinet Keita, 1,2,a Nicole Vidal, 1,a, Abdoulaye Toure, 2,3 Mamadou Saliou Kalifa Diallo, 2 N’Fally Magassouba, 4 Sylvain Baize, 5,6 Mathieu Mateo, 5,6 Herve Raoul, 7 Stephane Mely, 7 Fabien Subtil, 8,9 Cécé Kpamou, 2 Lamine Koivogui, 3 Falaye Traore, 3 Mamadou Saliou Sow, 2,10 Ahidjo Ayouba, 1 Jean Francois Etard, 1 Eric Delaporte, 1 and Martine Peeters 1, ; for the PostEbogui Study Group 1 TransVIHMI, IRD/INSERM/Monpellier University, Montpellier, France, 2 Centre de Recherche et de Formation en Infectiologie de Guinée, Université Gamal Abdel Nasser de Conakry, Conakry, Guinea, 3 Institut National de Santé Publique, Conakry, Guinea, 4 Laboratory of Virology, Conakry University, Projet de Recherche sur les Fièvres Hémorragiques en Guinée, Conakry, Guinea, 5 Institut Pasteur, Unit of Biology of Emerging Viral Infections - National Reference Center for Viral Hemorrhagic Fevers, Lyon, France, 6 Centre International de Recherche en Infectiologie, Lyon University - INSERM - ENS Lyon - CNRS, Lyon, France, 7 Laboratoire P4 Inserm-Jean Mérieux, US003 Inserm, Lyon, France, 8 Hospices Civils de Lyon, Service de Biostatistique, Lyon, France, 9 CNRS UMR 5558 Laboratoire de Biométrie et Biologie Évolutive, Équipe Biostatistique-Santé, Villeurbanne, France, 10 Donka National Hospital, Conakry, Guinea Background. With the increasing frequency and impact of Ebola virus disease (EVD) outbreaks illustrated by recent epidemics, a good understanding of the extent of viral persistance or ribonucleic acid (RNA) detection in body fuids from survivors is urgently needed. Methods. Ebola viral RNA shedding was studied with molecular assays in semen (n = 1368), urine (n = 1875), cervicovaginal fuid (n = 549), saliva (n = 900), breast milk (n = 168), and feces (n = 558) from EVD survivors in Guinea (PostEbogui cohort, n = 802) at a regular base period until 40 months afer inclusion. Results. Twenty-seven of 277 (9.8%) male survivors tested positive for Ebola RNA in at least 1 semen sample. Te probability of remaining positive for Ebola RNA in semen was estimated at 93.02% and 60.12% afer 3 and 6 months. Viral RNA in semen was more frequent in patients with eye pain (P = .036), joint pain (P = .047), and higher antibody levels to Ebola virus antigens (nucleo- protein [P = .001], glycoprotein [P = .05], and viral protein-40 [P = .05]). Ebola RNA was only rarely detected in the following body fuids from EVD survivors: saliva (1 of 454), urine (2 of 593), breast milk (2 of 168), cervicovaginal secretions (0 of 273), and feces (0 of 330). Ribonucleic acid was detected in breast milk 1 month afer delivery but 500 days afer discharge of Ebola treatment unit (ETU) in 1 woman who became pregnant 7 months afer discharge from the ETU. Conclusions. Te frequency and potential long-term presence of viral RNA in semen confrmed that systematic prevention measures in male survivors are required. Our observation in breast milk suggests that our knowledge on viral reservoir in immune- privileged sites and its impact are still incomplete. Keywords. body fuids; breast milk; Ebola; Guinea; semen. Since the first outbreak of Ebola virus disease (EVD) in 1976, in the Democratic Republic of Congo (DRC), 28 outbreaks have been reported across Africa [1]. In general, Ebola virus (EBOV) outbreaks remained restricted to rural or semirural areas and with a limited number of victims. The most important outbreak occurred between December 2013 and March 2016, when the virus spread from a rural area in southeast Guinea to the neigh- boring countries of Liberia and Sierra Leone and reached the capital cities from each country. More than 28 000 individuals became infected, and at least 11 000 people died [2]. The overall frequency of EBOV outbreaks seems to increase; for example, 3 independent outbreaks occurred between May 2017 and July 2018 in the DRC, and 2 of them also reached major cities [3, 4]. The actual outbreak that started in August 2018 in North Kivu has reached several densely populated cities, and today more than 3200 cases have already been identified, with mortality rate of approximately 60% [5]. Te major route of human-to-human transmission of EBOV is direct contact with infected body fuids from symptomatic or deceased patients. However, episodes of EBOV re-emergence and unusual transmission chains have been reported in the West African outbreak [6–10]. Te majority of these episodes were associated with viral persistence in semen, but transmission chains through other body fuids (breast milk, cervicovagianl fuids) are also suspected [7, 9, 11–13]. Te persistence of EBOV in body fuids was already shown in the outbreak from Downloaded from https://academic.oup.com/ofid/article-abstract/6/12/ofz482/5614563 by guest on 03 June 2020