Variation of prevalence of malaria, parasite density and the multiplicity of Plasmodium falciparum infection throughout the year at three different health centers in Brazzaville, Republic of Congo.

BACKGROUND: In the Republic of Congo, hot temperature and seasons distortions observed may impact the development of malaria parasites. We investigate the variation of malaria cases, parasite density and the multiplicity of Plasmodium falciparum infection throughout the year in Brazzaville. METHODS: From May 2015 to May 2016, suspected patients with uncomplicated malaria were enrolled at the Hôpital de Mfilou, CSI « Maman Mboualé¼, and the Laboratoire National de Santé Publique. For each patient, thick blood was examined and parasite density was calculated. After DNA isolation, MSP1 and MSP2 genes were genotyped. RESULTS: A total of 416, 259 and 131 patients with suspected malaria were enrolled at the CSI «Maman Mboualé¼, Hôpital de Mfilou and the Laboratoire National de Santé Publique respectively. Proportion of malaria cases and geometric mean parasite density were higher at the CSI «Maman Mboualé¼ compared to over sites (P-value <0.001). However the multiplicity of infection was higher at the Hôpital de Mfilou (P-value <0.001). At the Laboratoire National de Santé Publique, malaria cases and multiplicity of infection were not influenced by different seasons. However, variation of the mean parasite density was statistically significant (P-value <0.01). Higher proportions of malaria cases were found at the end of main rainy season either the beginning of the main dry season at the Hôpital de Mfilou and the CSI «Maman Mboualé¼; while, lowest proportions were observed in September and January and in September and March respectively. Higher mean parasite densities were found at the end of rainy seasons with persistence at the beginning of dry seasons. The lowest mean parasite densities were found during dry seasons, with persistence at the beginning of rainy seasons. Fluctuation of the multiplicity of infection throughout the year was observed without significance between seasons. CONCLUSION: The current study suggests that malaria transmission is still variable between the north and south parts of Brazzaville. Seasonal fluctuations of malaria cases and mean parasite densities were observed with some extension to different seasons. Thus, both meteorological and entomological studies are needed to update the season's periods as well as malaria transmission intensity in Brazzaville.

Diverse RNA viruses of arthropod origin in the blood of fruit bats suggest a link between bat and arthropod viromes.

Bats host diverse viruses due to their unique ecology, behavior, and immunology. However, the role of other organisms with which bats interact in nature is understudied as a contributor to bat viral diversity. We discovered five viruses in the blood of fruit bats (Hypsignathus monstrosus) from the Republic of Congo. Of these five viruses, four have phylogenetic and genomic features suggesting an arthropod origin (a dicistrovirus, a nodavirus, and two tombus-like viruses), while the fifth (a hepadnavirus) is clearly of mammalian origin. We also report the parallel discovery of related tombus-like viruses in fig wasps and primitive crane flies from bat habitats, as well as high infection rates of bats with haemosporidian parasites (Hepatocystis sp.). These findings suggest transmission between arthropods and bats, perhaps through ingestion or hyperparasitism (viral infection of bat parasites). Some "bat-associated" viruses may be epidemiologically linked to bats through their ecological associations with invertebrates.

No polymorphisms in K13-propeller gene associated with artemisinin resistance in Plasmodium falciparum isolated from Brazzaville, Republic of Congo.

BACKGROUND: In the Republic of Congo, artemisinin-based combinations have been recommended for the treatment of uncomplicated malaria since 2006. However, the emergence of resistant parasites again these combinations in Southeast Asia is a threat for the control of this disease, especially in sub-Saharan Africa where the weight of the disease is important. Indeed, polymorphisms in Plasmodium falciparum K13-propeller gene have been involved in variations of drug sensitivity of Plasmodium falciparum to artemisinin-based combinations. The aim of the current study is to determine the prevalence of mutations of this gene in isolates collected in three health centers in Brazzaville. METHODS: From May 2015 to May 2016, a total of 131, 259 and 416 samples from patients with suspected malaria were collected at the Laboratoire National de Santé Publique, Hôpital de Mfilou, and the CSI «Maman Mboualé¼ respectively. After DNA isolation, genotyping and sequencing of Plasmodium falciparum K13-propeller were performed in positive Plasmodium falciparum isolates identified after msp-2 gene genotyping. RESULTS: All 806 samples collected were msp-2 genotyped and Plasmodium falciparum infections were confirmed in 287 samples with 43, 85, 159 samples from Laboratoire National de Santé Publique, Hôpital de Mfilou, and the CSI «Maman Mboualé¼ respectively. Of these 287 msp-2 positives samples, K13-propeller nested PCR products were successfully obtained from 145 (50.52%) isolates and sequences were generated from 127(87.58%) nested products. None of mutations that were associated with ACTs resistance in Southeast Asia were detected on the samples from three different study sites from Brazzaville. However, one mutation type was observed at position 578, where alanine was substituted by serine (A578S) in two isolates (1.57%, 2/127), those from the Hôpital de Mfilou. No mutation was found in isolates from the two other sites. CONCLUSION: The current study shows a very limited polymorphism in the K13-propeller gene in isolates from the Republic of Congo and K13 polymorphisms associate with ACT resistance are not present in this country. However, permanent and large surveillance of resistant parasite population using K13-propeller gene is recommended.

Evaluation of Routine Microscopy Performance for Malaria Diagnosis at Three Different Health Centers in Brazzaville, Republic of Congo.

BACKGROUND: In Republic of Congo, malaria diagnosis still widely relies on microscopy. We aimed to evaluate the performance of routine microscopy for malaria diagnosis at three different health centers in Brazzaville. METHODS: A total of 259, 416, and 131 patients with clinical signs of uncomplicated malaria were enrolled at the Hôpital de Mfilou, Centre de Santé Intégré "Maman Mboualé," and Laboratoire National de Santé Publique, respectively. Two thick blood smears were prepared for each patient, the first being examined by routine microscopists and the second by expert. RESULTS: At the Hôpital de Mfilou, sensitivity was 62.1% and specificity was 67.3%. Positive and negative predictive values were 55.6% and 72.9%, respectively. At the Centre de Santé Intégré "Maman Mboualé," sensitivity was 94.2% and specificity was 33.6%. Positive and negative predictive values were 50% and 89.1%, respectively. At the Laboratoire National de Santé Publique, sensitivity and specificity were high with 91.7% and 94.9%, respectively. Positive and negative predictive values were 64.7% and 99.1%, respectively. CONCLUSION: The performance of routine malaria microscopy in Brazzaville remains inaccurate with large variations among different health centers. Therefore, repeated training including supervision and evaluation would improve routine malaria diagnosis for better management of malaria in Brazzaville, the Republic of Congo.

Detection of novel RNA viruses from free-living gorillas, Republic of the Congo: genetic diversity of picobirnaviruses.

Most of the emerging infectious diseases reported so far originated in wildlife. Therefore, virological surveillance of animals and particularly great apes is of great interest to establish the repertory of viruses associated with healthy hosts. This will further help to identify the emergence of new viruses and predict the possibility of interspecies transmission. In this study, we performed shotgun viral metagenomics on stool samples collected from seventeen free-living wild gorillas from the Republic of the Congo. The analysis revealed the presence of novel RNA viruses (picobirnaviruses, partitivirus, and Picornavirales (posa-like and dicistrovirus-like viruses)). Among these, picobirnavirus-related sequences were abundantly covered in the stools. Based on genetic variations both in capsid and RdRp proteins of picobirnaviruses, at least 96 variants were identified and most of them were novel. Among the 96, 22 variants had a nearly complete genome or segment. A comprehensive sequence analysis identified a potential new genogroup/genetic cluster and the presence of a short linear amino acid motif (ExxRxNxxxE) in a hypothetical protein. The sequence analysis of posa-like virus and dicistrovirus showed that these two viruses were novel members in the respective viral families. In conclusion, the identification of novel RNA viruses and their genetic diversity increases our knowledge about viruses that are associated with stools of wild gorillas and contributes to the initiatives in the search for potential emerging zoonotic viruses.

Syphilis and HIV infections among pregnant women attending antenatal clinics in Republic of Congo.

INTRODUCTION: HIV and syphilis during pregnancy remain a public health concern especially in developing countries. Pregnant women attending antenatal clinics sites for the first time between September and December 2011 and who accepted to participate in the study were enrolled. The objective was to estimate the syphilis and HIV infection rate in this population. METHODS: A study was conducted in 44 selected ANCs from 12 departments (5 urban and 7 rural). Pregnant women who accepted to participate in the study, attending selected sentinel ANCs sites for the first time between September and December 2011 were enrolled. To detect HIV antibodies, two consecutive ELISA assays were used (Genscreen Ultra HIV Ag/Ac, (BioRad, France) and Enzygnostic Intergral II (Siemens, GMBH, Marbug-Germany). In case of discordant results, the Western blot test II, HIV1 and 2 (Bio-Rad, Marne la Coquette, France) was used as the reference method. The RPR (Bio-Scan, Karnataka, India) test was performed to detect syphilis infection. The RPR positive results were confirmed using the TPHA test (Biotech, Cambridge, UK). Data were analyzed using SPSS 17.0 software. RESULTS: A total of 2979 pregnant women attending ANCs were enrolled. The global HIV infection rate was estimated to be 3.6% (CI: 95%; 3.0-4.4). As expected, HIV prevalence was significantly higher in women aged above 25 years (4.4% (3.4-5.6), p = 0.026) and those attending urban ANCs (5.04%, p < 0.01). Also, women living in the urban area are more at risk to be infected (5.04 VS 2.38, p < 0.01). The RPR test was positive in 117 pregnant women (3.92%). The risk for syphilis occurrence was significantly higher among the single women compared to the married ones (4.4% VS 2.7%; p < 0.01). It was also estimated that the HIV and syphilis coinfection occurred in 22 cases (0.73%). CONCLUSION: The prevalence's of syphilis and HIV were relatively low. Marital status and sentinel site location were a risk factor associated with HIV and syphilis infections respectively. Therefore, substantial effort is needed to reinforce prevention strategies in this population to prevent mother-to-child and further horizontal transmissions of these infections.

HIV-1 genetic diversity and primary drug resistance mutations before large-scale access to antiretroviral therapy, Republic of Congo.

BACKGROUND: In this work, we investigated the genetic diversity of HIV-1 and the presence of mutations conferring antiretroviral drug resistance in 50 drug-naïve infected persons in the Republic of Congo (RoC). Samples were obtained before large-scale access to HAART in 2002 and 2004. METHODS: To assess the HIV-1 genetic recombination, the sequencing of the pol gene encoding a protease and partial reverse transcriptase was performed and analyzed with updated references, including newly characterized CRFs. The assessment of drug resistance was conducted according to the WHO protocol. RESULTS: Among the 50 samples analyzed for the pol gene, 50% were classified as intersubtype recombinants, charring complex structures inside the pol fragment. Five samples could not be classified (noted U). The most prevalent subtypes were G with 10 isolates and D with 11 isolates. One isolate of A, J, H, CRF05, CRF18 and CRF37 were also found. Two samples (4%) harboring the mutations M230L and Y181C associated with the TAMs M41L and T215Y, respectively, were found. CONCLUSION: This first study in the RoC, based on WHO classification, shows that the threshold of transmitted drug resistance before large-scale access to antiretroviral therapy is 4%.

A study on HIV, Syphilis, and Hepatitis B and C virus infections among female sex workers in the Republic of Congo.

BACKGROUND: Female Sex Workers (FSWs) are considered to be at high risk for transmission of Sexually Transmitted Infections (STIs) and are defined as a priority of the national HIV/AIDS response in the Republic of Congo (RoC). However, no data are available regarding STIs in this group. This study aimed to determine the prevalences of HIV, syphilis and hepatitis B and C among FSWs in five cities in the country. METHODS: A cross-sectional study was conducted from November 2nd 2011 to May 15th 2012. Participants were recruited in Brazzaville, Pointe-Noire, Dolisie, Nkayi and Pokola using a respondent-driven sampling method. RESULTS: A total of 805 FSWs were recruited with an average age of 28.31 ± 9.15 years. The overall prevalences of HIV, syphilis, HBV and HCV were 7.50%, 2.20%, 4.20% and 0.70%, respectively. The age groups 35-39 (20.51% [0%-36.93%], p = 0.0057) and greater than 40 years (16.67% [0%-34.93%], P = 0.016) were positively associated with behaviors at high risk of HIV infection. For syphilis, the most infected age group was the one greater than 40 years, at 6.25% ([1.06% -72.37%] p = 0.04). Pointe-Noire was the most infected city for syphilis and HBV, with 5.15% (p = 0.0061) and 4.22% (p˂0.001), respectively. No risk factors were associated with HCV infection. FSWs practicing in mobile prostitution sites had a significantly higher infection rate (2.1% [0%-11.09%] p = 0.04). CONCLUSION: This study shows that the prevalence of HIV and other STIs in FSWs is high. Therefore, a combination of individual and structural interventions could reduce the risk of an STI "reservoir" among this population.

Head Lice of Pygmies Reveal the Presence of Relapsing Fever Borreliae in the Republic of Congo.

BACKGROUND: Head lice, Pediculus humanus capitis, occur in four divergent mitochondrial clades (A, B, C and D), each having particular geographical distributions. Recent studies suggest that head lice, as is the case of body lice, can act as a vector for louse-borne diseases. Therefore, understanding the genetic diversity of lice worldwide is of critical importance to our understanding of the risk of louse-borne diseases. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report the results of the first molecular screening of pygmies' head lice in the Republic of Congo for seven pathogens and an analysis of lice mitochondrial clades. We developed two duplex clade-specific real-time PCRs and identified three major mitochondrial clades: A, C, and D indicating high diversity among the head lice studied. We identified the presence of a dangerous human pathogen, Borrelia recurrentis, the causative agent of relapsing fever, in ten clade A head lice, which was not reported in the Republic of Congo, and B. theileri in one head louse. The results also show widespread infection among head lice with several species of Acinetobacter. A. junii was the most prevalent, followed by A. ursingii, A. baumannii, A. johnsonii, A. schindleri, A. lwoffii, A. nosocomialis and A. towneri. CONCLUSIONS/SIGNIFICANCE: Our study is the first to show the presence of B. recurrentis in African pygmies' head lice in the Republic of Congo. This study is also the first to report the presence of DNAs of B. theileri and several species of Acinetobacter in human head lice. Further studies are needed to determine whether the head lice can transmit these pathogenic bacteria from person to another.

Inflammatory status hepatic enzymes and serum creatinine in HIV-, HIV+ and HIV-TB co-infected adult Central Africans.

BACKGROUND AND AIM: Human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome is a major public health issue in Africa. The objective of this study was to determine which of isolated HIV-infection, isolated naive pulmonary tuberculosis (PTB), or naive HIV-PTB coinfection was more harmful to inflammatory, hepatic, and renal functions. METHODS: This cross-sectional study was undertaken among ten patients with isolated HIV infection, ten patients with isolated naive HIV infection, ten patients with isolated PTB and 32 patients with HIV-PTB coinfection, with the aim of determining which group had the highest levels of oxidative stress and hepatic and renal dysfunction markers. Serum aminotransferase (AST), alanine transferase (ALT), gamma-glutamyl transferase (GGT), and creatinine measurements were compared across the three groups of patients, who were managed from admission in the pulmonology division of the Brazzaville Teaching Hospital, Congo. RESULTS: HIV patients had the highest levels of ALT, GGT, and creatinine before and after adjusting for age and sex. Adjusted levels of AST, ALT, GGT, and creatinine were higher in HIV-PTB coinfection patients than in sero-negative PTB patients. CONCLUSION: There is a significant association between HIV infection and increase in concentration of ALT, GGT, and creatinine.

Biomarkers of oxidative stress and personalized treatment of pulmonary tuberculosis: emerging role of gamma-glutamyltransferase.

Background. The objectives were (i) to evaluate the impact of acute pulmonary tuberculosis (PTB) and anti-TB therapy on the relationship between AST, ALT, and GGT levels in absence of conditions related to hepatotoxicity; (ii) to evaluate the rate and the time of alterations of AST, ALT, and GGT. Design and Methods. A prospective followup of 40 adults (21 males; mean age of 34.7 ± 5.8 years) with active PTB on initial phase and continuation phase anti-TB. Results. Only 3% (n = 1) developed a transient and benign ADR at day 30 without interruption of anti-TB treatment. Within normal ranges, GGT decreased significantly from day 0 to day 60, while AST and ALT increased significantly and respectively. During day 0-day 60, there was a significant, negative, and independent association between GGT and AST. Conclusion. The initial two months led to significant improvement of oxidative stress. Values of oxidative markers in normal ranges might predict low rate of ADR.

[Lipid, lipoproteins and atherogenesis profiles in sickle cell disease among Central African patients]. / Évaluation du profil lipoprotéique et du risque athérogène chez les drépanocytaires homozygotes et hétérozygotes de Brazzaville.

Lipid and lipopproteins disorders are well established in sera from sickle cell disease (SCD) patients out of Central Africa. The present case-control study was conducted to compare serum levels of total cholesterol (TC), HDL-C, triglycerides (TG), LDL-C and TC/HDL-C ratio (atherogenic index) from SCD homozygotes (SS) in steady state, SCD heterozygotes (AS) and controls (AA) in Brazzaville, Congo. Significant reductions of TC and LDL-C vs. increase in TG were reported in SS. However, significant decrease in HDL-C and increase in atherogenic index were observed in AS. We recommend prevention of oxidative stress, dyslipidemia and atherosclerosis in SCD using hygiene-diet measures. Only longitudinal studies in large populations will provide pathophysiological basis of lipid and lipoproteins disorders in SCD.

Report of the 4th International Conference on Envenomations by Snakebites and Scorpion Stings in Africa, Dakar, April 25-29, 2011.

The authors present a summary of the proceedings and the recommendations of the the 4th International Conference on Envenomations by Snakebites and Scorpion Stings in Africa, held from April 25th to 29th in Dakar. After a two day training workshop for Senegalese health personnel on the most relevant aspects of the management of envenomations, about 270 participants met to share their experiences in the field. Nearly a hundred oral and poster contributions concerning the epidemiology of snakebites and scorpion stings in Africa, the composition and action of venoms, as well as the manufacture and use of antivenoms, were presented and discussed. The last day was devoted to an institutional debate joining experts, representatives of national health authorities and concerned professionals (physicians, pharmacists, nurses and traditional healers) as well as members of the pharmaceutical industry, to discuss and elaborate a set of recommendations. It was agreed that it is necessary to improve knowledge of the epidemiological situation by case reporting. Quality control of anivenoms and procedures for their registration at the level of national health authorities should aim at improving the distribution of safe and effective antivenoms in peripheral health centers, which bear the heaviest burden of cases. It was also recommended that adequate training of health personnel in all aspects of medical management of envenomations should constitute a priority. Finally, financing mechanisms to ensure an equitable distribution of resources must be sought, as well as the constitution of a network of African experts were discussed at length.

HIV-1 subtypes and recombinants in the Republic of Congo.

To document the actual genetic diversity of HIV-1 strains in the Republic of Congo, 114 HIV-1 positives persons were sampled in 2003 and 2004 after their informed consent. They were attending the teaching hospital, the reference health center in Makelekele, Brazzaville and the regional hospital centers in Pointe-Noire, Gamboma and Ouesso. A total of 104 samples were genetically characterized by direct sequencing of the p24 gag region and 80 were also subtyped in the V3-V5 env region. The genetic subtype distribution of the Congolese strains showed the predominance of subtype A (36.5% and 32.5% in gag and env, respectively) and G (30.8% and 21.25%), whereas subtype D strains represented 12.5% and 15%. Subtypes C, F, H, J, K and the CRFs-01, -02, -05 -06, and also the recently characterized CRF18 were seen at lower rates. Finally, 4.8% (gag) and 6.25% (env) of the strains could not be classified. Moreover, a high intra-subtype diversity was observed in our study. Among 70 strains which have been characterized in the two genomic regions, 14 (20%) appeared to be unique recombinants. These data show a high genetic variability in the Republic of Congo, where all the subtypes have been documented together with certain subsubtypes and several CRFs.

A novel simian immunodeficiency virus from black mangabey (Lophocebus aterrimus) in the Democratic Republic of Congo.

In order to understand primate lentivirus evolution, characterization of additional simian immunodeficiency virus (SIV) strains is essential. Here, an SIV from a black mangabey (Lophocebus aterrimus) originating from the Democratic Republic of Congo was analysed phylogenetically. The monkey had cross-reactive antibodies against human immunodeficiency virus type 1 (HIV-1) and HIV-2. The viral pol region sequence was amplified by nested PCR and sequence analysis confirmed that it was related to known SIV sequences. This is the first report to characterize genetically an SIV from the monkey genus Lophocebus. Phylogenetic analysis of the pol region revealed that this novel SIV, designated SIVbkm, fell into the SIVsyk and SIVgsn virus group, containing viruses isolated from the genus Cercopithecus, and suggests that cross-species transmission has occurred between species of the genera Lophocebus and Cercopithecus.

Genetic diversity of HIV type 1 in Likasi, southeast of the Democratic Republic of Congo.

To investigate the prevalence of subtypes A and C, and the existence of recombinants of both subtypes in the southeast of the Democratic Republic of Congo (DRC), blood samples were collected from 27 HIV-infected individuals in Likasi, located in an area bordering close to Zambia, and analyzed phylogenetically. Out of the 24 strains with a positive PCR profile for pol-IN and env-C2V3, 15 (62.5%) had a discordant subtype or CRF designation: one subtype A/G (pol/env), four A/U (unclassified), three G/A, one G/CRF01, three H/A, one J/C, one CRF02 (G)/A, and one U/A. Nine (37.5%) strains had a concordant subtype or CRF designation: five subtype A, two C, one D, and one CRF02/G. The remaining three samples negative for PCR with env-C2V3 primers used in this study were further analyzed with env-gp41 primers and revealed the presence of two profiles: two J/J (pol-IN/env-gp41) and one C/G. These data highlight the presence of a high proportion (16/27, 59.3%) of recombinant strains and a low prevalence (4.1 and 7.4%) of subtype C based on env-C2V3 and pol-IN analyses, respectively, in Likasi. In addition, this is the first report that CRF02_AG exists in DRC, though the epidemiological significance of the existence of CRF02_AG in DRC remains unknown.