The Wood equation allows consistent fitting of individual antibody-response profiles of Zika virus or SARS-CoV-2 infected patients.

Antibody kinetic curves obtained during a viral infection are often fitted using aggregated patient data, hiding the heterogeneity of individual humoral immune responses. Individual antibody responses can be modeled using the Wood equation and grouped according to their profile. Such modeling takes into account several important kinetic parameters, such as the day when antibody detection becomes positive [daypos], the day of the maximal response [daymax], the maximum antibody level [levelmax], and the day when antibody detection becomes negative [dayneg]. Potential associations between these profiles and studied factors can then be tested.

Discrimination of 15 Amazonian Anopheline Mosquito Species by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.

Precise identification of anopheline species is paramount for incrimination of malaria vectors and implementation of a sustainable control program. Anopheline mosquitoes are routinely identified morphologically, a technique that is time-consuming, needs high level of expertise, and prone to misidentifications especially when considering Amazonian species. The aim of this study was therefore to develop a DNA-based identification technique to supplement traditional morphological identification methods for the discrimination of anopheline mosquitoes collected in French Guiana. The internal transcribed spacer 2 (ITS2) region of ribosomal DNA (rDNA) for anopheline species was amplified by polymerase chain reaction (PCR), and digested with AluI/MspI restriction enzymes. PCR-restriction fragments length polymorphism (RFLP) assay was compared to sequencing of the ITS2 region for validation. Fifteen Anopheles species have shown distinct PCR-RFLP profiles. A concordance of 100% was obtained when identification by PCR-RFLP was compared to sequencing of ITS2. A high throughput, fast, and cost-effective PCR-RFLP assay has been developed for unambiguous discrimination of fifteen anopheline mosquito species from French Guiana including primary and suspected secondary malaria vectors.

Leishmaniavirus genetic diversity is not related to leishmaniasis treatment failure.

OBJECTIVES: The outcome of American tegumentary leishmaniasis (ATL) may depend on the presence of the Leishmania RNA virus (LRV). This virus may be involved in treatment failure. We aimed to determine whether genetic clusters of LRV1 are involved in this therapeutic outcome. METHODS: The presence of LRV1 was assessed in 129 Leishmania guyanensis isolates from patients treated with pentamidine in French Guiana. Among the 115 (89%) isolates found to carry LRV1, 96 were successfully genotyped. Patient clinical data were linked to the LRV data. RESULTS: The rate of treatment failure for LRV1-positive isolates was 37% (15/41) versus 40% (2/5) among LRV1-negative isolates (p 0.88). Concerning LRV1 genotypes, two predominant LRV1 groups emerged, groups A (23% (22/96)) and B (70% (67/96)). The treatment failure rate was 37% (3/8) for group A and 45% (9/20) for group B (p 0.31). DISCUSSION: Neither the presence nor genotype of LRV1 in patients with L. guyanensis seemed to correlate with pentamidine treatment failure.

Risk of daytime transmission of malaria in the French Guiana rain forest.

Between 2008 and 2014, there were 1070 malaria cases reported in French Guiana among members of the armed forces. Most of the malaria outbreaks investigated were multifactorial and followed missions conducted at illegal gold mining sites. For example, a malaria outbreak occurred in September 2013, three weeks after the deployment of 15 soldiers at Dagobert, which is such a site. The attack rate was 53%, with seven Plasmodium vivax infections and one coinfection with both Plasmodium vivax and Plasmodium falciparum. Two months later, an entomological investigation in the field caught 321 anopheles by the human landing catch method. Among them, 282 were Anopheles darlingi. One specimen was PCR-positive for P. vivax, for an infection rate of 0.4% (1/282). In 15.7% of these cases, the An. darlingi was caught during the day. The existence of daytime biting activity by An. darlingi in the Guianese forest might play a key role in malaria outbreaks among military personnel. This finding requires that the Army Health Service adapt its recommendations concerning malaria prevention in French Guiana.

Lung nodule in French Guiana in a immunocompetent patient.

We report the case of an immunocompetent French soldier stationed in French Guiana, who developed symptomatic pulmonary histoplasmosis.

Spatial and Seasonal Dynamics of Anopheles Mosquitoes in Saint-Georges de l'Oyapock, French Guiana: Influence of Environmental Factors.

Little is known about the Anopheles fauna of Saint-Georges de l'Oyapock, a persistent malaria-endemic municipality in French Guiana. This study aimed to update the knowledge of local Anopheles diversity, and their ecology and role in malaria transmission. Sampling sessions were implemented between September 2013 and October 2014. Four species were identified from the 3,450 specimens collected: Anopheles darlingi Root, An. braziliensis, An. triannulatus s.l., and An. nuneztovari s.l. Anopheles darlingi was the predominant species. Its involvement in malaria transmission was suspected due to 1) its abundance, 2) the presence of a density peak during the malaria emergence period, and 3) a dynamic correlated with malaria cases observed two months later. Present and past studies show that the influence of environmental conditions on malaria vector dynamics is high, and may vary drastically according to the local context. This supports evidence that control strategies must be designed at fine scales.

[Role of primaquine in malaria control and elimination in French-speaking Africa]. / Place de la primaquine dans la lutte contre le paludisme en Afrique francophone.

Primaquine, an 8-aminoquinoline, is a relatively unknown and underutilized drug in French-speaking African countries. It acts against the liver stage parasites of all human malaria species, asexual blood stages of Plasmodium vivax and, to a lesser degree, Plasmodium falciparum; P. falciparum mature gametocytes, and P. vivax and Plasmodium ovale hypnozoites. Gastrointestinal disturbances are its most common side effects. The clinical utility of primaquine is limited due to its hematological side effects in individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency and other contraindications (pregnant woman, breastfeeding woman, infants less than 6 months old). In the light of the recent recommendations of the World Health Organization (WHO), we propose to examine how primaquine can be used in French-speaking Africa to improve malaria control and move towards malaria elimination. Two indications supported by the WHO are of relevance in Africa. First, artemisinin-based combination therapies and primaquine given as a single low dose (0.25 mg base/kg) are effective to kill asexual and sexual parasites of P. falciparum, are well-tolerated, and have very little risk even in mild to moderate G6PD-deficient patients. This strategy may be helpful to contain transmission in an area in Africa where P. falciparum malaria incidence has decreased considerably. There is an ethical concern in administering primaquine as a gametocytocide as it does not confer any direct benefit to the treated patient. However, the single low-dose primaquine is most likely associated with very low risk for adverse hematological effects, and WHO recommends its use even without prior G6PD testing. In our opinion, clinical studies including G6PD test should be conducted to assess the safety of low-dose primaquine in African patients. Second, primaquine is effective and necessary for radical treatment of P. vivax and P. ovale, but the standard 14-day treatment (0.25-0.5 mg base/kg/day) is not recommended in patients with G6PD deficiency. Prior G6PD testing is required before prescribing primaquine for radical treatment. The use of primaquine for radical treatment in patients without contraindications does not raise any major ethical problem since the probability of relapse in patients who do not receive anti-hypnozoite treatment can be relatively high and each relapse can cause or aggravate anemia, especially in children. In our opinion, patients with mild or moderate G6PD deficiency should not be treated with primaquine at present. Further clinical studies are necessary to define the role of this drug for radical treatment in G6PD-deficient African patients. Without primaquine, the eventual elimination of P. vivax and P. ovale malaria appears to be very difficult. Updated epidemiological data on G6PD, Duffy antigen, and the current distribution of and burden due to P. vivax and P. ovale are required for a rational use of primaquine in the African continent. Moreover, clinical studies on primaquine are required in Africa.

Zika virus infections.

Since its discovery in 1947 in Uganda, the Zika virus (ZIKV) remained in the shadows emerging in 2007 in Micronesia, where hundreds of dengue-like syndromes were reported. Then, in 2013-2014, it was rife in French Polynesia, where the first neurological effects were observed. More recently, its arrival in Brazil was accompanied by an unusually high number of children with microcephaly born to mothers infected with ZIKV during the first trimester of pregnancy. In 2016, the World Health Organization declared ZIKV infection to be a public health emergency and now talks about a ZIKV pandemic. This review aims to summarize the current knowledge about ZIKV infection, successively addressing its transmission, epidemiology, clinical aspects, diagnosis, treatment, and prevention before discussing some perspectives.

[Delayed diagnosis of Plasmodium falciparum in a soldier in Uganda: false-positive rapid diagnostic test associated with reduced repeats in pfhrp2]. / Retard de diagnostic d'un accès palustre à Plasmodium falciparum chez un militaire en Ouganda : négativité des tests de diagnostic rapide associée à un polymorphisme de l'antigène HRP2.

Rapid diagnostic tests (RDTs) are the best alternative for malaria diagnosis where a microscopic examination cannot be performed. We report here the first case of P. falciparum (false-negative) misdiagnosis in a soldier stationed in Uganda, associated with a reduced number of repeats in the P. falciparum histidine-rich protein 2 gene (pfhrp2). This gene was subsequently sequenced to determine the reason for the discordance between the RDT results and the later microscopic examination. Ten repeats of the type 2 motif AHHAHHAAD and four repeats of the type 7 motif AHHAAD were found. This isolate belongs to the group of non-sensitive parasites (<43 repeats) that are not detected by HRP2 RDTs. This inappropriate case management could have been fatal for the patient. This case confirms the problem of negative RDT results in isolated situations and of basing a therapeutic strategy on these negative results. Investigations should be conducted in Uganda and other areas of Africa to determine the presence and the geographical spread of parasites with pfhrp2 gene deletion to ensure the best performance of RDTs.

Absence of association between ex vivo susceptibility to doxycycline and pftetQ and pfmdt copy numbers in Plasmodium falciparum isolates from Dakar, Senegal.

The objective of this study was to validate the use of pftetQ and pfmdt genes as molecular markers of decreased in vitro susceptibility to doxycycline in 113 Plasmodium falciparum isolates from Dakar, Senegal. The results show that copy numbers of pftetQ and pfmdt, estimated by TaqMan real-time PCR, are not significantly associated with reduced susceptibility to doxycycline in vitro; however, the number of samples with a high doxycycline IC(50) was likely to be too low to derive statistically significant results. Thus, no definitive conclusions could be drawn. The markers should be further tested by analysing more isolates.

[Molecular markers of Plasmodium falciparum drug resistance]. / Marqueurs moléculaires des résistances de Plasmodium falciparum aux antipaludiques.

The main Plasmodium falciparum genes known to be associated with drug resistance are pfcrt, pfmdr1, pfdhfr pfdhps, pfcytb, pfmrp, pfnhe1, pfmdt, pfserca and pftetQ. Molecular markers for resistance to chloroquine, amodiaquine, mefloquine, sulfadoxine-pyrimethamine, cycloguanil and atovaquone have been validated and used to predict the parasitological and clinical efficacy of these drugs (i.e. based on in vivo tests). These molecular markers have numerous advantages. They allow evaluation of large series of samples in less time than in vivo tests. Collection, transport and storage of samples are much easier than for in vitro tests. These markers can be used for epidemiological monitoring of resistance for an entire country as well as for prediction of therapeutic outcome for a single individual. Development of high-throughput molecular biology techniques, availability of the nucleotidic sequences of P. falciparum genomes, and close collaboration between fundamental researcher workers, clinical practitioners, and officials in charge of the national malaria control programs are major assets in the research and development of molecular markers for P. falciparum resistance to antimalarial drugs.

[Distinguishing Plasmodium falciparum populations for clinical research]. / Distinguer les populations de Plasmodium falciparum en recherche clinique.

The genetic diversity of Plasmodium falciparum is generally high. Study of this diversity is useful to differentiate the strains present in an individual before and after malaria treatment or to check if several individuals have been infected by the same parasites. Interpretation of the in vivo test recommended by the WHO to evaluate antimalarial drug efficacy requires distinguishing recrudescence from new infection when recurrent parasitemia suggests the possibility of therapeutic failure and antimalaria resistance. For this purpose, molecular biology techniques such as nested-PCRs are becoming increasingly available and can now be used in most endemic areas. An effort has been made to standardize P. falciparum genotyping carried out to distinguish recrudescence from new infection. Standardization has focused not only on genotyping methods but also on interpretation criteria. Compliance with these recommendations should improve the quality of clinical research and allow comparison of data from different centers.

[Proteomic analysis and parasitosis: principles and applications]. / Analyses protéomiques et parasitoses: principes et applications.

O'Farrel described a method allowing two-dimensional (2D) protein separation more than 30 years ago. Since then the original technique has made enormous progress. This progress has been accompanied by advances in mass spectrometry technology as well as various genome-sequencing programs. Today 2D electrophoresis has become the workhorse of proteomics, allowing resolution of complex structures containing thousands of proteins and providing a global view of the state of a proteome. This article presents the different steps and limitations of proteomic analysis: preparation of biological material, 2D electrophoresis, protein detection systems, and available tools for protein identification. Alternative proteomic approaches to 2D electrophoresis are also presented. A few applications are described as examples to illustrate the utility of proteomic analysis for studying the mechanisms underlying virulence, resistance to antimalarial therapies and immune response against pathologic agents.

[Cyclines and malaria]. / Cyclines et paludisme.

The emergence and rapid spread of Plasmodium falciparum resistance to various antimalarials compounds is gradually reducing the clinician's options for treating malaria and for adapting prophylaxis to each traveler and destination. In this context doxycycline is an increasingly useful alternative except in individuals with contraindications, mainly children under the age of eight and pregnant women. Already used successfully in association with quinine for treatment of malaria in areas with multiresistance, doxycline has also proven to be effective and well tolerated for prophylaxis of malaria. No resistance to doxycycline has been observed to date. Most reported prophylactic failures have been related to poor compliance during the month following return from the endemic zone. The mechanisms of action of doxycycline on the parasite are still unclear. Identification of the molecular targets of doxycycline would open the way for the design of more active structural analogues with longer half-life.

[Malaria vaccines: prospects and reality]. / Vaccins contre le paludisme: perspectives et réalité

The development of a malaria vaccine has been accelerating in the last ten years. The number of clinical trials has increased and some malaria antigens have been tested in endemic areas. No potential vaccine has yet shown sufficient and lasting efficacy to justify its inclusion in a public health program. However, trials have unambiguously shown that some level of anti-malaria clinical immunity can be achieved by vaccination, both in experimental and in field conditions. Advances in malaria vaccine development are presented.

In vitro inhibition of Chikungunya and Semliki Forest viruses replication by antiviral compounds: synergistic effect of interferon-alpha and ribavirin combination.

Chikungunya virus (CHIKV) and Semliki Forest virus (SFV) were used in our laboratory to screen active antiviral compounds against viruses of the Alphavirus genus. Antiviral activity was estimated by the reduction of the cytopathic effect of each alphavirus on infected Vero cells and by virus titer reduction. Cytotoxicity was evaluated by determining the inhibition of Trypan blue exclusion in confluent cell cultures and by the evaluation of the inhibitory effect on cell growth. With CHIKV and SFV, the selectivity indices of human recombinant interferon-alpha and iota-carrageenan were much higher than that of ribavirin, which has been previously investigated for its inhibitory effect on alphavirus infections. Compared to ribavirin, 6-azauridine was more effective against CHIKV and showed a similar antiviral activity against SFV. IFN-alpha2b, glycyrrhizin, 6-azauridine, and ribavirin caused a concentration-dependent reduction in the virus yield with CHIKV and SFV. Moreover, the combination of IFN-alpha2b and ribavirin had a subsynergistic antiviral effect on these two alphaviruses and should be evaluated for the treatment of these infections.