Detection of Plasmodium falciparum in Saliva and Stool Samples from Children Living in Franceville, a Highly Endemic Region of Gabon.

Due to the difficulty of obtaining blood samples, which is the invasive method that is currently used for the detection of Plasmodium spp., alternative diagnostic sampling methods that are effective and non-invasive are needed, particularly for long-term studies. Saliva and stool samples from malaria-infected individuals contain trace amounts of Plasmodium DNA and therefore could be used as alternatives. Malaria was screened using rapid diagnosis tests and confirmed via microscopy. Nested PCR tests targeting the Plasmodium falciparum-specific STEVOR gene were performed for blood, saliva and stool samples that were positive for malaria. Three hundred sixty-seven (367) children were enrolled and eighty (22.22%) were confirmed to be positive for malaria. Matched blood, saliva and stool samples were available for 35 children. By using blood smears as the gold standard for the diagnosis of malaria, our study indicates that Plasmodium DNA was more detectable in blood (100%) than in saliva (22.86%) and stools (14.29%). Applying qPCR to the STEVOR gene to detect Plasmodium falciparum DNA in saliva and stool samples cannot be considered as an alternative to the current malaria detection processes using blood specimens.

Pentaclethra eetveldeana Leaves from Four Congo-Brazzaville Regions: Antioxidant Capacity, Anti-Inflammatory Activity and Proportional Accumulation of Phytochemicals.

Oxidative stress and inflammation play a key role in the occurrence of neurodegenerative diseases. Traditionally, Pentaclethra eetveldeana leaves are used in dementia treatment. Therefore, this study aimed to evaluate the antioxidant and anti-inflammatory activities as well as the phytochemical composition of Pentaclethra eetveldeana leaves from four Congo-Brazzaville regions. The 1.2-diphenyl-1-picrylhydrazyl radical-scavenging, ß-carotene bleaching and molybdenum reduction assays were used to assess the antioxidant activity. The protein denaturation and erythrocyte membrane stabilization tests were used to analyze the anti-inflammatory activity. Phytochemical screening, the quantification of polyphenols by spectrophotometry, as well as the determination of extraction yields were carried out. It was found that the extracts reduced molybdenum; furthermore, compared to ascorbic acid, they showed better antiradical activity and inhibited lipid peroxidation. Moreover, globally, the membrane-stabilizing power of the aqueous extracts was superior or comparable to diclofenac, while the same extracts were less effective for the inhibition of denaturation. All of the aqueous extracts contained polyphenols, saponins, alkaloids, anthraquinones, reducing sugar and cardiotonic glycosides. The total polyphenols, tannins and proanthocyanidins are produced proportionally from one region to another. Finally, the leaves from Brazzaville and Boundji contain flavonols, while those from Makoua and Owando contain flavones. Thus, Pentaclethra eetveldeana leaves contribute to traditional dementia treatment through their antioxidant and anti-inflammatory properties.

Severe malaria in children and adolescents in Southeast Gabon.

INTRODUCTION: Malaria remains a significant public health problem in sub-Saharan Africa. Child mortality due to severe malaria remains high in developing countries despite improvements in malaria management and a better understanding of its pathophysiology. To address the lack of epidemiological studies on severe malaria in Gabon, this study describes the epidemiological aspects of severe malaria in rural, semi-rural, and urban areas of southeast Gabon. METHODS: Demographic, clinical, and laboratory data for children and adolescents aged 0-18 years were collected in 2019 from hospital records at three health facilities in southeastern Gabon. The patients included in the study were positive for P falciparum malaria diagnosed by microscopy with at least one of the malaria severity criteria. RESULTS: Severe malaria accounted for 18.8% (667/3552) of malaria cases. Children aged 0-5 years accounted for 71.8% (479/667) of all severe malaria cases. Adolescents over 15 years of age were the least affected by severe malaria with 4.2% (28/667). Across the study, severe anemia (49.0%, 327/667), convulsions (43.0%, 287/667), respiratory distress (5.1%, 34/667), and altered consciousness (4.8%, 32/667) were the most frequent clinical signs of severe malaria in children. Franceville was the locality most affected by severe malaria with 49.2% (328/667), followed by Koulamoutou with 42.0% (280/667) and Lastourville with 8.8% (59/667). Convulsions (50.6%, 166/328) and coma (6.1%, 20/328) were more frequent in children living in urban areas. In contrast, severe anemia (56.7%, 186/339) and jaundice (6.8%, 23/339) were more common in children living in semi-rural areas. CONCLUSION: Severe malaria is more prevalent in urban areas in regions with a high malaria transmission intensity. However, in this study, the epidemiological characteristics of severe malaria were similar in the three settings (urban, rural, and semi-rural areas) despite different levels of urbanization. Nevertheless, the various signs of severity were more frequent in Franceville, an urban area. Children under 5 years of age remain the most vulnerable age group.

Severe malaria in Gabon: epidemiological, clinical and laboratory features in Amissa Bongo Hospital of Franceville.

BACKGROUND: Malaria is the most deadly parasitic disease and continues to claim more than a half million of deaths across the world each year, mainly those of under-fives children in sub-Saharan Africa. The aim of this study was to determine the epidemiological, clinical and laboratory features of patients with severe malaria at the Centre Hospitalier Régional Amissa Bongo (CHRAB), a referral hospital in Franceville. METHODS: It was an observational descriptive study conducted at CHRAB over 10 months. All admitted patients at the emergency ward of all ages presenting with positive test to falciparum malaria diagnosed by microscopy and rapid test with clinical signs of severe illness describe by World Health Organization were enrolled. RESULTS: During this study, 1065 patients were tested positive for malaria, of them 220 had severe malaria. Three quarters (75.0%) were less than 5 years of age. The mean time to consultation was 3.5 ± 1 days. The most frequent signs of severity on admission were dominated by neurological disorders 92.27% (prostration 58.6% and convulsion 24.1%), followed by severe anemia 72.7%, hyperlactatemia 54.6%, jaundice 25% and respiratory distress 21.82%.The other forms such as hypoglycemia, haemoglobinuria, renal failure were found in low proportions < 10%. Twenty-one patients died, coma (aOR = 15.54, CI 5.43-44.41, p < 0.01), hypoglycemia (aOR = 15.37, CI 2.17-65.3, p < 0.01), respiratory distress (aOR = 3.85, CI 1.53-9.73, p = 0.004) and abnormal bleeding (aOR = 16.42, CI 3.57-104.73, p = 0.003) were identified as independent predictors of a fatal outcome. Anemia was associated with decreased mortality. CONCLUSION: Severe malaria remains a public health problem affecting mostly children under 5 years. Classification of malaria helps identify the most severely ill patients and aids early and appropriate management of the severe malaria cases.

Intermittent preventive treatment and malaria amongst pregnant women who give birth at the Centre Hospitalier Régional Paul Moukambi de Koula-Moutou in southeastern Gabon.

BACKGROUND: Evaluating malaria control strategies for pregnant women is essential. The objective of this study was to determine the factors influencing antenatal care (ANC) visit attendance, complete intermittent preventive treatment with sulfadoxine-pyrimethamine (IPTp-SP) and its impact on the health of pregnant women and their newborn babies living in semi-urban and rural areas of southeastern Gabon. METHODS: This transversal study was performed at the Centre Hospitalier Régional Paul Moukambi de Koula-Moutou (CHRPMK). Information regarding age, frequency of prenatal consultations, obstetric history, use of malaria control measures, use of IPTp-SP, malaria diagnostic of women and their newborns, were collected: (i): from birth registers from 1 January, 2018 to 31 December, 2019 and, (ii): a questionnaire from January to April 2020. RESULTS: In total, 1,851 and 323 pregnant women were included during the first and the second sub-set of study, respectively. In the first sub-set of data, the mean age was 26.18 ± 7.02 years and 96.54% (1,787/1,851) of pregnant women had attended ANC service but 54.45% had complete ANC visit attendance (at least 4 ANC). The complete ANC visit was linked with age (p < 0.001) and profession (p < 0.001). The complete IPTp-SP (at least 3 doses) was 58.87%. Complete IPTp-SP was linked to profession (aOR = 1.49, 95% CI [1.04-2.18], p < 0.001), ANC visit (aOR = 0.176, 95% CI [0.14-0.22], p < 0.034) and age (p = 0.03). Birth weight was higher for babies whose mothers had received complete IPTp-SP (p < 0,001) but the Apgar score was not influenced by the use of IPTp-SP (p = 0.71). In the second sub-set of data, the prevalence of plasmodial infection was 3.10% (95% IC [1.21-5]) and Plasmodium falciparum was responsible for 100% of infections. The prevalence of plasmodial infection was the same for all age groups (p = 0.69), gravidity (p = 0.13) and domestic control measures (p > 0.05). A low birth weight was statistically linked to the mother's plasmodial infection (p < 0.01). Furthermore, plasmodial infection was statistically linked to premature birth (p < 0.001). CONCLUSIONS: It was observed that attendance of women to ANC service and a complete IPTp-SP course is insufficient.

Malaria research in the Central African Republic from 1987 to 2020: an overview.

BACKGROUND: The national malaria control policy in the Central African Republic (CAR) promotes basic, clinical, and operational research on malaria in collaboration with national and international research institutions. Preparatory work for the elaboration of National Strategic Plans for the implementation of the national malaria control policy includes developing the research component, thus requiring an overview of national malaria research. Here, this survey aims to provide an inventory of malaria research as a baseline for guiding researchers and health authorities in choosing the future avenues of research. METHODS: Data sources and search strategy were defined to query the online Medline/PubMed database using the "medical subject headings" tool. Eligibility and study inclusion criteria were applied to the selected articles, which were classified based on year, research institute affiliations, and research topic. RESULTS: A total of 118 articles were retrieved and 51 articles were ultimately chosen for the bibliometric analysis. The number of publications on malaria has increased over time from 1987 to 2020. These articles were published in 32 different journals, the most represented being the Malaria Journal (13.73%) and the American Journal of Tropical Medicine and Hygiene (11.76%). The leading research topics were drug evaluation (52.94%), expatriate patients (23.54%), malaria in children (17.65%), morbidity (13.7%), and malaria during pregnancy (11.76%). The publications' authors were mainly affiliated with the Institut Pasteur of Bangui (41%), the French Military Medical Service (15.5%), and the University of Bangui (11.7%). Collaborations were mostly established with France, the UK, and the USA; some collaborations involved Switzerland, Austria, Pakistan, Japan, Sri Lanka, Benin, Cameroun, Ivory Coast, and Madagascar. The main sources of research funding were French agencies (28.6%) and international agencies (18.3%). Most studies included were not representative of the whole country. The CAR has the capacity to carry out research on malaria and to ensure the necessary collaborations. CONCLUSION: Malaria research activities in the CAR seem to reflect the priorities of national policy. One remaining challenge is to develop a more representative approach to better characterize malaria cases across the country. Finally, future research and control measures need to integrate the effect of COVID-19.

Knowledge, Attitude and Practice during the COVID-19 Pandemic in South-East Gabon.

COVID-19 is an emerging respiratory disease; it was declared a global pandemic by the World Health Organization. This survey aimed to describe the knowledge, attitudes, perception and practices of the Gabonese public towards COVID-19. This study was performed on 1016 participants. All participants filled in the questionnaire voluntarily, reporting demographic characteristics and answering questions assessing their level of knowledge, attitudes, perceptions and practice towards COVID-19. Among participants, there were 535 men and 476 women. The mean age of the participants was 33.2 ± 16.7 years old. Almost all participants (98.1%) said that they had heard about COVID-19 but only 2.8% knew the pathogenic agent responsible for COVID-19. More than 80% knew that the disease could be transmitted by greeting infected people (87.3%), kissing an infected person (90.0%), touching an infected doorknob (83.5%) and attending meetings (83.9%). The mean knowledge score was higher among younger than older participants, higher among participants living in urban areas than those living in rural areas and higher among participants with higher levels of education than those with lower levels of education. In general, respondents had good knowledge of COVID-19 and a positive attitude towards using protective measures; however, there were differences according to gender, age group, place of residence, professional group and level of education.

Differential Prevalences of Pfmdr1 Polymorphisms in Symptomatic and Asymptomatic Plasmodium falciparum Infections in Lastoursville: A Rural Area in East-Central Gabon.

PURPOSE: Plasmodium falciparum malaria remains a major public health challenge in sub-Saharan Africa. Plasmodium falciparum drug resistance mediated by polymorphisms in the Pfmdr1 gene contributes to the persistence of the disease on the African continent. This study investigated P. falciparum infection features and differences in the Pfmdr1 genotypes between symptomatic and asymptomatic malaria cases in a rural area in east-central Gabon. PATIENTS AND METHODS: A total of 875 children aged from 5 to 185 months were screened for P falciparum infection using Optima-IT® rapid diagnostic tests and standard microscopy. Pfmdr1 polymorphisms at codons 86, 184 and 1246 were investigated using PCR-RFLP. RESULTS: Among the 448 P. falciparum-infected children, 57.08% (n=250) were symptomatic and 42.92% (n=198) were asymptomatic (p < 0.0001). In a sub-set of 79 isolates, the Pfmdr1 wild-type N86 was more prevalent in symptomatic (100%) than in asymptomatic infections (70.7%) (p=0.007). The mutant 86Y and mixed 86N/Y genotypes were observed only in asymptomatic infections. The Y184 and 184F genotype prevalences (39.1% vs 19.4% and 60.9% vs 80.6%, respectively) were not significantly different between the two groups (p=0.097). The prevalence of the wild-type D1246 differed significantly between symptomatic (10.3%) and asymptomatic (100%) (p < 0.0001). The NFD and YFD haplotypes were more prevalent in asymptomatic than in symptomatic infections [(61.9% vs 31%; p=0.005) and (16.7% vs 0.0%; p=0.01)], whereas the NYD and YYD haplotypes were not significantly different between the two groups [(21.4% vs 14.3%, p=0.39) and (0.0% vs 7.1%, p=0.24)]. CONCLUSION: Our results confirm a high transmission of P. falciparum infection in rural Gabon, with a high prevalence of asymptomatic carriage. The higher prevalences of wild-type N86 in symptomatic infections and of D1246 in asymptomatic infections suggest a pathogenicity associated with polymorphisms in Pfmdr1. These results highlight the need to monitor the efficacy of artemisinin-based combination therapies in Gabon.

Malaria in children and women of childbearing age: infection prevalence, knowledge and use of malaria prevention tools in the province of Nyanga, Gabon.

BACKGROUND: There is little information on the social perception of malaria and the use of preventative measures in Gabon, especially in rural areas. Adequate knowledge of malaria prevention and control can help in reducing the burden of malaria among vulnerable groups, particularly pregnant women and children under 5 years old living in malaria-endemic settings. This study was designed to assess the prevalence of malaria and the knowledge and attitude towards this disease in households in Nyanga Province. METHODS: A cross-sectional study was conducted to assess malaria knowledge, prevention practices and prevalence of the malaria infection in five departments of Nyanga Province. Plasmodial infection was diagnosed in children ≤ 5 years of age and women aged 15-49 years using rapid diagnostic tests. A questionnaire was administered randomly to women aged 15-49 years and to the parents or guardians of children aged ≤ 5 years in 535 households during a 2-week period in March 2018. Overall, the respondents' socio-demographic characteristics, knowledge of malaria, malaria prevention practices and malaria prevalence were evaluated and compared across the five departments. RESULTS: Data from a total of 1,307 participants were included in this study, including 631 women of childbearing age (61 of them pregnant) and 676 children. Practically the entire (97.7%) interviewed population had heard about malaria and attributed the cause of malaria to a mosquito bite (95.7%). This survey revealed that the reported rate of reported bed-net use was 73.3%. The study observed an average malaria parasite prevalence of 13.9%. All departmental capitals of Nyanga Province had a significant level of malaria infection except for Mayumba where no plasmodial infection was found. CONCLUSION: High malaria prevalence is found in the departmental capital cities of Nyanga Province. This study reveals that respondents have a high knowledge of the malaria symptoms, its mode of transmission and preventive measures. Despite this high level of knowledge of the disease and its preventive measures, the incidence of malaria remains relatively high in this rural community highlighting the need for other types of interventions.

Epidemiology and molecular characterization of the re-emerging measles virus among children and adults in the Haut-Ogooue, Gabon.

BACKGROUND: Measles is one of the most infectious diseases with a high mortality rate worldwide. It is caused by the measles virus (MeV) which is a single stranded RNA virus with genetic diversity based on the nucleoprotein gene, including 24 genotypes. In Gabon, several outbreaks occurred in the past few years, especially in 2016 in Libreville and Oyem. A surveillance network of infectious diseases highlighted a measles outbreak which occurred in the south of Gabon from April to June 2017. METHODS: Clinical specimens of urine, blood, throat and nasal swabs were collected in the two main cities of the Haut-Ogooue province, Franceville and Moanda. Virological investigations based on real-time polymerase chain reaction for molecular diagnosis and conventional PCR for genotype identification were done. RESULTS: Specimens were collected from 139 suspected measles patients. A total of 46 (33.1%) children and adults were laboratory-confirmed cases among which 16 (34.8%) were unvaccinated, 16 (34.8%) had received one dose, and 11 (23.9%) had received two doses of the measles vaccine. Phylogenetic analysis revealed that all the sequences of the nucleoprotein gene belonged to genotype B3. CONCLUSIONS: Measles infection was more commonly confirmed among those with one recorded dose compared to suspect cases with none, unknown or two recorded doses. The molecular characterization of the strains showed the circulation of the B3 genotype which is endemic on the African continent, thirty years after the B2 genotype was described in an outbreak in Libreville, the capital of Gabon. These findings highlight that surveillance and molecular investigation of measles should be continued in Gabon.

High Level of Specific Anti-Plasmodium Falciparum Merozoite IgG1 Antibodies in Rural Asymptomatic Individuals of Dienga, South-Eastern Gabon.

Plasmodium falciparum merozoite antigens (PfMAgs) play an essential role in the development of immunity to malaria. Currently, P. falciparum: protein 113 (Pf 113), apical membrane antigen 1 (AMA1), erythrocyte binding antigens (EBA175), and reticulocyte binding protein homologue 5 (RH5) are among the most PfMAgs studied. A comparative analysis of naturally acquired antibodies against these antigens in children would increase our knowledge about the development of protective immunity. Analysis of antibodies to Pf113, PfAMA1, PfEBA175, and PfRH5 was conducted in rural population during 2013 and 2014. Both prevalence and levels of total IgG anti-PfAMA1 were higher than that of IgG anti-PfEBA175, anti-PfRH5, and anti-Pf113. Seroconversion to PfAMA1 and PfEBA175 occurred moderately in young children and reached to the maximum in adolescent and in adults. High prevalence of IgG anti-Pf113 was observed in young children of 3 to 6 years old in 2013. The four antigens were recognized by IgG 1, 2, 3, and 4 antibodies from a large proportion of the subjects, and all of them induced high levels of specific IgG1 against PfAMA1, PfEBA175, fewer by Pf113 and PfRH5. Many asymptomatic children had specific IgG1 recognizing multiple antigens, and these IgG1 antibodies could be associated with a reduced risk of developing malaria symptoms.

Comparative Antibody Responses Against three Antimalarial Vaccine Candidate Antigens from Urban and Rural Exposed Individuals in Gabon.

The analysis of immune responses in diverse malaria endemic regions provides more information to understand the host's immune response to Plasmodium falciparum. Several plasmodial antigens have been reported as targets of human immunity. PfAMA1 is one of most studied vaccine candidates; PfRH5 and Pf113 are new promising vaccine candidates. The aim of this study was to evaluate humoral response against these three antigens among children of Lastourville (rural area) and Franceville (urban area). Malaria was diagnosed using rapid diagnosis tests. Plasma samples were tested against these antigens by enzyme-linked immunosorbent assay (ELISA). We found that malaria prevalence was five times higher in the rural area than in the urban area (p < 0.0001). The anti-PfAMA1 and PfRh5 response levels were significantly higher in Lastourville than in Franceville (p < 0.0001; p = 0.005). The anti-AMA1 response was higher than the anti-Pf113 response, which in turn was higher than the anti-PfRh5 response in both sites. Anti-PfAMA1 levels were significantly higher in infected children than those in uninfected children (p = 0.001) in Franceville. Anti-Pf113 and anti-PfRh5 antibody levels were lowest in children presenting severe malarial anemia. These three antigens are targets of immunity in Gabon. Further studies on the role of Pf113 in antimalarial protection against severe anemia are needed.

Immunoglobulin response to the low polymorphic Pf113 antigen in children from Lastoursville, South-East of Gabon.

Pf113 is a P. falciparum putatively GPI-anchored protein that has been so far localized at the surface of merozoites, suggesting it could interact with RBC surface during merozoite invasion. Previous studies conducted in Papua New Guinea and in Kenya have revealed that this protein is recognized by natural antibodies in individuals living in malaria-endemic areas and is associated with protective immunity in malaria, further supporting the potential of Pf113 for the development of anti-malaria vaccines. However, in Central Africa, no study on the immunogenicity of this protein has been conducted. Here, we report the characterization of the Pf113 immune response in 103 children by Enzyme-Linked Immunoabsorbent Assay (ELISA), using a recombinant form of Pf113 expressed in Escherichia coli, together with the study of the Pf113 polymorphism, after amplification and sequencing of 40 field isolates. Data showed that almost 51% of the studied individuals had positive antibody responses to the recombinant Pf113 protein, and that IgG subclass response was dominated by IgG3 (84%) followed by IgG1 (50%). Surprisingly the prevalence of IgG4 was 92%. In addition, gene analysis in field isolates from this region indicated that Pf113 was not highly polymorphic, in particular regarding high-activity binding peptides (HABPs). Our data reinforce the idea that Pf113 may be considered for inclusion in multicomponent blood-stage vaccines.

Malaria in urban, semi-urban and rural areas of southern of Gabon: comparison of the Pfmdr 1 and Pfcrt genotypes from symptomatic children.

BACKGROUND: Artesunate-amodiaquine (AS-AQ) and artemether-lumefantrine (AL) are first- and second-line treatments for uncomplicated Plasmodium falciparum malaria in Gabon. AL remains highly efficacious, but its widespread use has led to molecular selection of the NFD haplotype on Pfmdr1 and K76 in Pfcrt. In this study, plasmodial infection characteristics and the distribution of the Pfmdr1 and Pfcrt genotypes involved in reduced efficacy of artemisinin-based combination therapy (ACT) were investigated in four Gabonese localities. METHODS: A cross-sectional study was conducted in the paediatric units of rural (Lastourville and Fougamou), semi-urban (Koula-Moutou) and urban (Franceville) areas. Malaria was diagnosed with the rapid diagnostic test Optimal-IT(®) and confirmed by blood smear. Pfmdr1 codons 86, 184 and 1246 and Pfcrt codon 76 were genotyped by PCR-RFLP and sequencing. RESULTS: Among 1129 included children, the prevalence of plasmodial infection was 79.5 % at Lastourville, 53.6 % at Fougamou, 36.1 % at Koula-Moutou, and 21.2 % at Franceville. The prevalence was significantly higher among children over 60 months of age in both semi-urban (p = 0.01) and urban (p = 0.004) areas. The prevalence of Pfmdr1 wild-type N86 differed significantly between Lastourville (57.8 %) and Koula-Moutou (45.4 %) (p = 0.039). No difference in 184F-carrying parasites was found between Lastourville (73.8 %), Fougamou (81.6 %), Koula-Moutou (83.2 %), and Franceville (80.6 %) (p = 0.240). The prevalence of wild-type D1246 was significantly different between Lastourville (94.1 %), Koula-Moutou (85.6 %) and Franceville (87.3 %) (p = 0.01). The frequency of wild-type K76 was not significantly different across the four sites: Lastourville (16.5 %), Fougamou (27.8 %), Koula-Moutou (17.4 %), and Franceville (29.4 %) (p = 0.09). The mixed genotypes were only found in Lastourville and Franceville. The NFD, YFD and NYD haplotypes were mainly Lastourville (46.6, 25.8, 14.0 %), Fougamou (45.5, 9.1, 42.4 %), Koula-Moutou (35, 6.7, 40.4 %), and Franceville (40.0, 16.0, 32.0 %). CONCLUSION: This study shows an increase in the prevalence of childhood plasmodial infection in Gabon according to the low socio-economic level, and a high frequency of markers associated with AL treatment failure. Close monitoring of ACT use is needed.