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Plasmodium Ovale Cryptic Plasmodium ovale concurrent with mixed Plasmodium falciparum and Plasmodium malariae infection in two children from Central African Republic Cynthia Bichara, Philippe Flahaut, Damien Costa, Anne-Lise Bienvenu, Stéphane Picot, Gilles Gargala To cite this version: Cynthia Bichara, Philippe Flahaut, Damien Costa, Anne-Lise Bienvenu, Stéphane Picot, et al.. Cryp- tic Plasmodium ovale concurrent with mixed Plasmodium falciparum and Plasmodium malariae in- fection in two children from Central African Republic. Malaria Journal, BioMed Central, 2017, 16, pp.339. 10.1186/s12936-017-1979-5. hal-02316787 HAL Id: hal-02316787 https://hal.archives-ouvertes.fr/hal-02316787 Submitted on 15 Oct 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Bichara et al. Malar J (2017) 16:339 DOI 10.1186/s12936-017-1979-5 Malaria Journal CASE REPORT Open Access Cryptic Plasmodium ovale concurrent with mixed Plasmodium falciparum and Plasmodium malariae infection in two children from Central African Republic Cynthia Bichara1, Philippe Flahaut2, Damien Costa1, Anne‑Lise Bienvenu3, Stephane Picot3,4 and Gilles Gargala1* Abstract Background: Since several malaria parasite species are usually present in a particular area, co-infections with more than one species of Plasmodium are more likely to occur in humans infected in these areas. In many mixed infections, parasite densities of the cryptic species may be low and often not recognized in clinical practice. Case presentation: Two children (3 and 6 years old) adopted recently from Central African Republic were admitted to hospital because of intermittent fever. Thin blood smears stained with Giemsa showed Plasmodium falciparum and Plasmodium malariae co-infection for both children at admission. They were both treated with atovaquone-proguanil combination for 3 days. At day 7, both thin blood smears examination remained negative but at day 28, thin blood smear was positive for P. malariae trophozoites and for Plasmodium ovale for the girl and her brother, respectively. Samples collected at day 1 and day 28 were submitted to real-time PCR showing the presence of the three parasite species (P. falciparum, P malariae and P. ovale) in admission blood samples from the two children and only P. ovale at day 28. Conclusions: Twenty-eight days follow-up after treatment led to detection of a third parasite species in the blood of these two patients suggesting covert co-infection and a delayed appearance of one cryptic species following treat‑ ment. Concurrently infecting malaria species could be mutually suppressive, with P. falciparum tending to dominate other species. These observations provide more evidence that recommendations for treatment of imported malaria should take into account the risk of concurrent or cryptic infection with Plasmodium species. Clinicians and biologists should be aware of the underestimated frequency of mixed infections with cryptic species and of the importance of patient follow-up at day 28. Future guidelines should shed more light on the treatment of mixed infection and on the interest of using artemisinin-based combinations for falciparum and non-falciparum species. Keywords: Malaria, Plasmodium, Cryptic, Diagnosis, Follow-up Background is rare in clinical practice, and probably underestimated Mixed-species malaria infections have received spo- by microscopy. Expert microscopists are uncommon in radic attention [1], and the trend toward malaria control clinical laboratory, and distinguishing the species from in some endemic areas should reduce the occurrence of young trophozoite forms of Plasmodium parasites is concurrent infection with more than one species of Plas- sometimes difcult [2]. Te life-threatening P. falcipa- modium. However, diagnostic of these mixed-infections rum is obviously taking most of the attention consider- ing the potential consequences of a wrong diagnosis, and *Correspondence: gilles.gargala@univ‑rouen.fr other species are sometimes scanned with less intensity. 1 Department of Parasitology‑Mycology, Rouen University Hospital, In case of mixed infection, the concurrent species is 76 000 Rouen, France mostly below the threshold of microscopy. For example, Full list of author information is available at the end of the article © The Author(s) 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Bichara et al. Malar J (2017) 16:339 Page 2 of 4 the ratio of Plasmodium malariae/P. falciparum was coagulation test result for the prothrombin time–inter- described as extremely low in a study conducted in Ivory national normalized ratio was slightly normal. Ultrasono- Coast and the protective efect of lengthy subclinical P. graphic abdominal examination reported a splenomegaly malariae infections was suspected [3]. Moreover, the (100 mm, and lack of deep lymph nodes. liver stage of Plasmodium vivax and Plasmodium ovale, Tin blood smears stained with Giemsa showed P. falci- which are responsible for relapses, are not detectable [1]. parum and P. malariae co-infection for both children (P. Te delayed appearance of a cryptic species after suc- falciparum 1.3% + P. malariae 0.14% and P. falciparum cessful treatment of the frst diagnosed species has been 0.03% + P. malariae 0.15% for girl and boy, respectively). reported in area endemic for P. falciparum and P. vivax A rapid diagnostic test (Palutop4 +, AllDiag, Strasbourg, malaria [4]. France) showed three positive bands for the control, It has been known from decades that antagonism Plasmodium spp. (pLDH), and P. falciparum (pHRPII) between species may lead to a rapid decrease of circulat- markers. ing parasites, such as P. malariae when inoculated with P. Both children received 1 adult co-formulated tablet of vivax in an attempt to treat neurosyphilis [5]. Malarone, containing 250 mg atovaquone and 100 mg Triple species infection are known, but remain rare in proguanil hydrochloride, per day for 3 days. Te tablets the literature. Two parallel cases of triple infections (P. were administered during the meal without addition falciparum and P. malariae, followed by P. ovale delayed of fat. Te boy weighed 19 kg, the girl weighed 14.5 kg infection) in two adopted children from Central African and the administered weight-based dose of atovaquone/ Republic are presented here to document the risk of late proguanil was 10/5 and 17/7 mg/kg/day for the boy and reappearance and the need for clinical and parasitologi- girl respectively. Tey did not vomit and no other adverse cal follow-up. Tese cases focused more light on the risk events were noted during treatment. Both children of treating each malaria species by drugs with diferent received iron syrup supplementation and paracetamol. efcacy. Almost complete parasitaemia clearance was obtained at day 3 (P. falciparum <0.01% P. malariae <0.01% and Case report + P. falciparum 0% + P. malariae 0.02%, for girl and boy Two children (3 and 6 years old) were admitted to hospi- respectively). Tese fgures are not uncommon because tal because of intermittent fever and suspected malaria. atovaquone-proguanil is known to act relatively slowly. Tey have been adopted recently and came back from Apyrexia was obtained at day 2 and patients were dis- Central African Republic a week previously. charged on day 5 and did not return until scheduled Te 6-year-old boy presented with a temperature of appointments for control of parasitaemia on day 7. 37.9 °C, a systolic murmur in mitral focus, and a hepato- At day 7, both thin blood smears examination remained splenomegaly. Te haemoglobin level was 6.7 g/dL, the negative, while few picnotic parasites were detected on mean corpuscular volume was 65 fL, and the platelet thick blood smears. Surprisingly, at day 28, thin blood count was 216 G/L. Biochemical tests showed a normal smear from the girl was positive and revealed <0.01% liver function, increased serum levels of lactate dehy- P. malariae trophozoites while P. ovale (<0.01%) was drogenase (291 U/L, reference range 135–250 U/L) and detected from her brother’s blood. To confrm the species C-reactive protein (73, reference value <5). A coagulation identifcation, the samples collected at day 1 and day 28 test result for the prothrombin time–international nor- were submitted to real-time PCR as previously described malized ratio was slightly prolonged (14.7 s with a ratio at [6]. Molecular diagnosis demonstrated the presence of 67%, reference range 12.5 s with a ratio at 75–100%). An the three parasite species (P. falciparum, P. malariae and ultrasonographic abdominal examination showed hepa- P. ovale) in D0 samples from both children, and only P. tomegaly (107 mm thickness along the mammary line, ovale at day 28. reference value for this age 90 mm), and splenomegaly Both children were still apyretic and didn’t show any (115 mm, reference range 70–96 mm), and lack of deep symptoms of infection. Considering the suspected para- lymph nodes. An ultrasonographic heart examination sitological failure of the atovaquone-proguanil treat- reported no abnormalities. ment against P. ovale, patients were given a total dose of Te 3-year-old girl, presented with a temperature of 25 mg/kg of chloroquine over 3 days.
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