Antimicrobial Plants Used by Fang Populations and Phytochemical Profiling of Erismadelphus exsul.

Gabon has a rich flora, many species of which are used in traditional medicine. However, little research has been carried out on this wealth. An ethnopharmacological survey in the Fang language was carried out among traditional practitioners to collect antimicrobial medicinal plants. Phytochemical profiling of ethanolic and methanolic extracts from Erismadelphus exsul Mildbr leaves was carried out using HPLC-ESI-Q/TOF and a molecular network approach. Antibacterial activity was assessed with disk diffusion and microdilution methods, antioxidant activity via DPPH and FRAP methods, and in vitro cell viability via Cell Counting Kit-8. A total of 21 medicinal plants were collected, grouped into 10 families, of which the Fabaceae is the most represented. Erismadelphus exsul was chosen for chemical and biological studies due to its citation frequency (RCF = 0.59) and the absence of previous phytochemical studies. These studies revealed 4 major families of natural compounds and annotated 19 compounds for the first time. The crude leaf extract showed significant antioxidant and antibacterial activity. Cytotoxicity studies showed that the leaves were not cytotoxic, unlike the bark. This study underlines the importance of preserving the ancestral knowledge of the Fang populations, while showing promising results for Erismadelphus exsul.

Targeted Isolation of Hemitheion from Mostuea brunonis, a Proposed Biosynthetic Intermediate of Theionbrunonines.

Hemitheion (1), a new sulfur-containing vobasane-type indole alkaloid, was isolated, together with three known compounds, vobasine (2), gelsedine (3), and gelsemicine (4), from the alkaloid extract of the stems of Mostuea brunonis Didr. (Gelsemiaceae). Compound 1 could be straightforwardly isolated. Its structure was elucidated by a combination of spectroscopic methods. Besides corresponding to a formerly postulated biosynthetic intermediate toward theionbrunonines, hemitheion (1) stands among the few monomeric vobasanes lacking an oxygen at C-3. Hemitheion (1) showed moderate antiplasmodial activity in the micromolar range against the strain FcB1 of Plasmodium falciparum and no cytotoxic activity against the MRC-5 cell line at 20 µM.

A genome scan for Plasmodium falciparum malaria identifies quantitative trait loci on chromosomes 5q31, 6p21.3, 17p12, and 19p13.

BACKGROUND: Genome-wide studies have mapped several loci controlling Plasmodium falciparum mild malaria and parasitaemia, only two of them being significant at the genome level. The objective of the present study was to identify malaria resistance loci in individuals living in Burkina Faso. METHODS: A genome scan that involved 314 individuals belonging to 63 families was performed. Markers located within chromosomes 6p21.3 and 17p12 were genotyped in 247 additional individuals belonging to 55 families. The linkage and the association of markers with parasitaemia and mild malaria were assessed by using the maximum-likelihood binomial method extended to quantitative trait linkage and the quantitative trait disequilibrium test, respectively. RESULTS: Multipoint linkage analysis showed a significant linkage of mild malaria to chromosome 6p21.3 (LOD score 3.73, P = 1.7 10-5), a suggestive linkage of mild malaria to chromosome 19p13.12 (LOD score 2.50, P = 3.5 10-4), and a suggestive linkage of asymptomatic parasitaemia to chromosomes 6p21.3 (LOD score 2.36, P = 4.9 10-4) and 17p12 (LOD score 2.87, P = 1.4 10-4). Genome-wide family-based association analysis revealed a significant association between three chromosome 5q31 markers and asymptomatic parasitaemia, whereas there was no association with mild malaria. When taking into account 247 additional individuals, a significant linkage of asymptomatic parasitaemia to chromosome 17p12 (LOD score 3.6, P = 2 10-5) was detected. CONCLUSION: A new genome-wide significant malaria locus on chromosome 17p12 and a new suggestive locus on chromosome 19p13.12 are reported. Moreover, there was evidence that confirmed the influence of chromosomes 5q31 and 6p21.3 as loci controlling mild malaria or asymptomatic parasitaemia.

Cytotoxicity and Identification of Antibacterial Compounds from Baillonella toxisperma Bark Using a LC-MS/MS and Molecular Networking Approach.

Baillonella toxisperma is a medicinal plant used in northern Gabon to treat microbial diseases. It is a plant well-known by local populations, but very few studies have focused on the molecules responsible for the antibacterial activities of B. toxisperma. This study proposes a dereplication strategy based on molecular networking generated from HPLC-ESI-Q/TOF data, allowing investigation of the molecules responsible for the antibacterial activity of B. toxisperma. From this strategy, eighteen compounds were putatively identified. All of these compounds belonged mainly to five families of natural compounds, including phenylpropanolamines, stilbenes, flavonoids, lignans and phenolic glycosides. The chemical study carried out from the bark of B. toxisperma allowed us to identify, for the first time, compounds such as resveratrol and derivatives, epicatechin, epigallocatechin and epigallocatechin gallate. In addition, antibacterial activity (diffusion method and microdilution) and cytotoxicity (Cell Counting Kit-8 (CCK-8 Assay)) in vitro were evaluated. The crude ethanolic extract, as well as the fractions of B. toxisperma, showed significant antibacterial activity. However, the ethanolic fractions F2 and F4 presented high antibacterial activity compared to the crude extract. Cytotoxicity studies on colon-cancer cells (Caco-2) and human keratinocyte cells (HaCaT) showed moderate cytotoxicity in both cell types. This study clearly shows the therapeutic potential of the ethanolic extract of the bark of B. toxisperma and provides information on the phytochemical composition and bioactive compounds of the plant.

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.

Molecular Characterization of Canine Parvovirus Variants CPV-2a and CPV-2c, Associated with Vaccinated Dogs at Libreville, Gabon.

The first detection of canine parvovirus type-2 (CPV-2) was in the early 1970s, when it was known to cause severe gastroenteritis in dogs. However, it has evolved over the years into CPV-2a within 2 years, into CPV-2b after 14 years, into CPV-2c after 16 years and more recently CPV-2a-, 2b- and 2c-like variants reported in 2019, with a global distribution. Reports on the molecular epidemiology of this virus are missing in most African countries. The report of clinical cases among vaccinated dogs in Libreville in Gabon triggered the execution of this study. The objective of this study was to characterize circulating variants from dogs showing clinical signs suggestive of CPV that were examined by a veterinarian. A total of eight (8) fecal swab samples were collected, and all had positive PCR results. Sequencing, Blast analysis and assembly of two whole genomes and eight partial VP2 sequences were performed, and the sequences submitted to GenBank. Genetic characterization revealed the presence of CPV-2a and CPV-2c variants with predominance of the former. Phylogenetically, the Gabonese CPVs formed distinct groups similar to Zambian CPV-2c and Australian CPV-2a sequences. The antigenic variants CPV-2a and CPV-2c have not yet been reported in Central Africa. However, these CPV-2 variants circulate in young, vaccinated dogs in Gabon. These results suggest additional epidemiological and genomic studies are required in order to evaluate the occurrence of different CPV variants in Gabon and effectiveness of the commercial vaccines used against protoparvovirus in the country.

The Antibiotics Used in Livestock and Their Impact on Resistance in Enterococcus faecium and Enterococcus hirae on Farms in Gabon.

The emergence of antibiotic resistance is a major concern around the world. The objective of this study was to investigate the antibiotics used in livestock and their impact on resistance in Enterococcus faecium and Enterococcus hirae on farms in Gabon. A structured questionnaire was used to collect information on the farms. Samples were collected from farms (n = 20) tested for Enterococcus by culture and isolation and were identified using a polymerase chain reaction (PCR) and sequencing. Antibiotic susceptibility was determined by the disc diffusion method on Mueller Hinton agar. The 20 farms included laying hens (6), swine (6), sheep (4) and cattle farms (4). Tetracycline was the most used antibiotic family (91%) and the most used prophylactic method (47%) for the treatment of animals. A total of 555 samples were collected and 515 (93%) Enterococcus spp. isolates of the genus were obtained. The prevalence of E. faecium and E. hirae were 10% and 8%, respectively. The isolates from E. faecium and E. hirae we found were related to clinical and human isolates in the NCBI database. E. faecium and E. hirae isolates showed a high resistance to tetracycline (69% and 65%) and rifampicin (39% and 56%). The tet(M) gene was detected in 65 tetracycline-resistant isolates with a large majority in hens (78% (21/27) and 86% (12/14) in E. faecium and E. hirae, respectively). The consumption of antibiotics favours the emergence of antibiotic resistance in animals in Gabon.

Investigating antibiotic resistance in enterococci in Gabonese livestock.

Background and Aim: The emergence of antibiotic resistance is a major problem worldwide. Antibiotics are often used to prevent or treat infections in livestock. This study aimed to investigate antibiotic resistance in enterococci in Gabonese livestock. Materials and Methods: We collected 174 animal samples (46 laying hens, 24 swine, 62 cattle, and 42 sheep) from farms in four provinces of Gabon. Bacterial strains belonging to the genus Enterococcus were obtained using selective media and polymerase chain reaction targeting the tuf gene. Antibiotic susceptibility was determined by the disk diffusion method on Mueller-Hinton agar. Results: Enterococci were present in 160 of the samples (97%), distributed as follows: laying hens (100%, 41/41), swine (100%, 22/22), small ruminants (88%, 37/42), and cattle (100%, 60/60). Resistance to cephalothin/cephalexin, streptomycin, and rifampicin (RIF) was high, and resistance to vancomycin (VAN), erythromycin, and tetracycline was moderate. A high diversity of resistance was found in Haut-Ogooué and Estuaire provinces. Laying hens and swine showed moderate levels of resistance to ciprofloxacin and penicillin, while sheep and cattle had high levels of resistance to RIF. All species showed a high level of resistance to VAN. We found various patterns of multiple resistances in the isolates, and the multiple resistance indexes ranged from 0.2 to 0.8. Conclusion: This study shows that livestock in Gabon can be considered potential reservoirs of resistance.

Combined LC-MS/MS and Molecular Networking Approach Reveals Antioxidant and Antimicrobial Compounds from Erismadelphus exsul Bark.

Erismadelphus exsul Mildbr bark is widely used in Gabonese folk medicine. However, little is known about the active compounds associated with its biological activities. In the present study, phytochemical profiling of the ethanolic extract of Erismadelphus exsul was performed using a de-replication strategy by coupling HPLC-ESI-Q/TOF with a molecular network approach. Eight families of natural compounds were putatively identified, including cyclopeptide alkaloids, esterified amino acids, isoflavonoid- and flavonoid-type polyphenols, glycerophospholipids, steroids and their derivatives, and quinoline alkaloids. All these compounds were identified for the first time in this plant. The use of molecular networking obtained a detailed phytochemical overview of this species. Furthermore, antioxidant (2,2-diphenyl-1-picryl-hydrazylhydrate (DPPH) and ferric reducing capacity (FRAP)) and in vitro antimicrobial activities were assessed. The crude extract, as well as fractions obtained from Erismadelphus exsul, showed a better reactivity to FRAP than DPPH. The fractions were two to four times more antioxidant than ascorbic acid while reacting to FRAP, and there was two to nine times less antioxidant than this reference while reacting to DPPH. In addition, several fractions and the crude extract exhibited a significant anti-oomycete activity towards the Solanaceae phytopathogen Phytophthora infestans in vitro, and, at a lower extent, the antifungal activity against the wheat pathogen Zymoseptoria tritici had growth inhibition rates ranging from 0 to 100%, depending on the tested concentration. This study provides new insights into the phytochemical characterization and the bioactivities of ethanolic extract from Erismadelphus exsul bark.

Vaginal Infections' Etiologies in South-Eastern Gabon - An Overview.

Introduction: Discomfort in women of childbearing age associated with vaginal infections, namely bacterial vaginosis (BV), aerobic vaginitis (AV), vulvovaginal candidiasis (VVC), and trichomoniasis (TV), represent a serious and ongoing gynecological complication throughout the world. Objective: This study aimed to investigate the etiologies of vaginal infections among outpatients in south-eastern Gabon. Methodology: A cross-sectional study was designed using participants referred directly by their treating doctor for a vaginal swab. Socio-demographic data were collected using a structured questionnaire. Microscopic examinations were used for TV and BV diagnostic. All vaginal swabs were cultured for AV and VVC isolates using standard microbiology methods. Results: A total of 573 women of reproductive age participated in the study. The most common identified vaginal infections were BV (62.8%) and AV (51.1%) followed by VVC (34.1%). No significant difference was observed for each etiology compared to socio-demographic data. Streptococcus B (23.9%), Staphylococcus aureus (17.7%), Klebsiella spp. (11.6%), and E. coli (5.8%) were the bacteria most associated with AV. A high incidence of non-C. albicans Candida (NCAC) strains causing vulvovaginitis were found. The prevalence of TV (2.1%) was low. Mixed infections had been common among participants. No association was found with TV and other vaginal infections, unlike others studies. The present study identified BV 228 (83.5%) and AV 227 (83.2%) as the main cause of mixed infections. The mixed infection AV-BV 113 (41.4%) was the most represented. Conclusion: Also that simultaneous AV-BV-VVC represented 69 (25.3%) of mixed infections. Molecular analyses would be needed to identify the key species commonly associated with these vaginal infections.

Fruit bats as reservoirs of Ebola virus.

The first recorded human outbreak of Ebola virus was in 1976, but the wild reservoir of this virus is still unknown. Here we test for Ebola in more than a thousand small vertebrates that were collected during Ebola outbreaks in humans and great apes between 2001 and 2003 in Gabon and the Republic of the Congo. We find evidence of asymptomatic infection by Ebola virus in three species of fruit bat, indicating that these animals may be acting as a reservoir for this deadly virus.

Frequency and diversity of trypanosomes in sheep and goats from Mongo County in South Gabon, Central Africa.

BACKGROUND AND AIM: Trypanosomosis is a major impediment to livestock farming in sub-Saharan Africa. It is a vector-borne disease caused by several species of protozoan parasites, namely, trypanosomes. The present study aimed to identify the diversity of trypanosome species infecting sheep and goats from Mongo County and to determine the frequency of these parasites. MATERIALS AND METHODS: This study was conducted on 286 trypanotolerant goats and sheep from Mongo regions located in South Gabon, using polymerase chain reaction. RESULTS: Analyses showed that the overall occurrence of trypanosomosis in small ruminants was 13.6% (39/286). Our results also showed that two factors, species and season, could affect the occurrence rate of Trypanosoma. A total of six Trypanosoma species were identified, two in sheep (Trypanosoma simiae and Trypanosoma theileri) and five in goats (Trypanosoma vivax, T. simiae, T. simiae Tsavo, Trypanosoma congolense, and Trypanosoma brucei), though Trypanosoma simiae was the most important species. Mixed infections were also found in goats (54.5%) and sheep (3.57%). CONCLUSION: Our study demonstrated that small ruminants could represent a reservoir of biodiversity for Trypanosoma parasites.

Phenylpropane as an Alternative Dearomatizing Unit of Indoles: Discovery of Inaequalisines A and B Using Substructure-Informed Molecular Networking.

Inaequalisines A and B (1 and 2), the first examples of hybrid alkylated phenylpropane monoterpene indole alkaloids, were isolated from the roots of Callichilia inaequalis, guided by the combined use of molecular networking and substructure annotation. Their structures, including absolute configuration, were elucidated by spectroscopic methods and ECD calculations. A possible biosynthetic pathway for 1 and 2 was postulated.

Theionbrunonines A and B: Dimeric Vobasine Alkaloids Tethered by a Thioether Bridge from Mostuea brunonis.

Theionbrunonines A and B (1 and 2), the first examples of monoterpene bisindole alkaloids linked by a thioether bridge, were isolated from the stems of Mostuea brunonis, guided by a molecular networking-based dereplication strategy. Their structures were elucidated by a combination of spectroscopic data and ECD calculations. A plausible biosynthetic pathway for 1 and 2 was postulated. Theionbrunonines A and B (1 and 2) showed moderate antiplasmodial activities in the micromolar range against the strain FcB1 of Plasmodium falciparum and no cytotoxic activity against the MRC-5 cell line at 20 µM.

Pleiokomenines A and B: Dimeric Aspidofractinine Alkaloids Tethered with a Methylene Group.

Pleiokomenines A and B (1 and 2), first examples of dimeric aspidofractinine alkaloids linked by a methylene bridge, were isolated from the stem bark of Pleiocarpa mutica. Their structures were elucidated by a combination of spectroscopic analyses and semisynthetic derivatization of pleiocarpinine (3) and a formaldehyde equivalent mediated by scandium trifluoromethanesulfonate. Pleiokomenines A (1) and B (2) showed moderate antiplasmodial activities in the micromolar range against the FcB1 strain.

Molecular identification of trypanosome species in trypanotolerant cattle from the south of Gabon.

The aim of this study was to provide information on trypanosome species infecting trypanotolerant cattle from southern Gabon. The study was conducted on 224 trypanotolerant cattle from three regions located in southern Gabon, using ITS1 primer-based PCR. Seventy-two (32%) N'dama cattle were found polymerase chain reaction (PCR) positive with trypanosomes. The overall prevalence of trypanosomosis was 57% (63/110), 4% (4/100), and 36% (5/14) in the Gala section of the Nyanga ranch, the Miyama ranch, and Ossiele, respectively. Trypanosoma congolense and Trypanosoma vivax were identified. In Gala section and Ossiele, T. congolense and T. vivax were found. In the Miyama ranch, only T. vivax was identified. Mixed infections were also found. The forest (9%) and savannah (63%) subgroups of T. congolense were identified. The presence of the two subgroups was detected in 16 out of 56 cattle (29%). T. congolense and T. vivax would appear to be the main agents responsible for bovine trypanosomosis in southern Gabon. Although trypanotolerant, N'dama cattle may serve as a reservoir, and this should be further studied. On the other hand, these trypanotolerant cattle can be reared in such tsetse infested areas, which gives them an advantage compared to other trypanosensitive breeds, and this shows that they represent a key factor in biodiversity which has to be promoted.

The natural history of Ebola virus in Africa.

Several countries spanning the equatorial forest regions of Africa have had outbreaks of Ebola hemorrhagic fever over the last three decades. This article is an overview of the many published investigations of how Ebola virus circulates in its natural environment, focusing on the viral reservoir, susceptible animal species, environmental conditions favoring inter-species transmission, and how the infection is transmitted to humans. Major breakthroughs have been made in recent years but many outstanding questions must be dealt with if we are to prevent human outbreaks by interfering with the viral life cycle.

Wild animal mortality monitoring and human Ebola outbreaks, Gabon and Republic of Congo, 2001-2003.

All human Ebola virus outbreaks during 2001-2003 in the forest zone between Gabon and Republic of Congo resulted from handling infected wild animal carcasses. After the first outbreak, we created an Animal Mortality Monitoring Network in collaboration with the Gabonese and Congolese Ministries of Forestry and Environment and wildlife organizations (Wildlife Conservation Society and Programme de Conservation et Utilisation Rationnelle des Ecosystemes Forestiers en Afrique Centrale) to predict and possibly prevent human Ebola outbreaks. Since August 2001, 98 wild animal carcasses have been recovered by the network, including 65 great apes. Analysis of 21 carcasses found that 10 gorillas, 3 chimpanzees, and 1 duiker tested positive for Ebola virus. Wild animal outbreaks began before each of the 5 human Ebola outbreaks. Twice we alerted the health authorities to an imminent risk for human outbreaks, weeks before they occurred.

Ebola virus antibody prevalence in dogs and human risk.

During the 2001-2002 outbreak in Gabon, we observed that several dogs were highly exposed to Ebola virus by eating infected dead animals. To examine whether these animals became infected with Ebola virus, we sampled 439 dogs and screened them by Ebola virus-specific immunoglobulin (Ig) G assay, antigen detection, and viral polymerase chain reaction amplification. Seven (8.9%) of 79 samples from the 2 main towns, 15 (15.2%) of 99 samples from Mekambo, and 40 (25.2%) of 159 samples from villages in the Ebola virus-epidemic area had detectable Ebola virus-IgG, compared to only 2 (2%) of 102 samples from France. Among dogs from villages with both infected animal carcasses and human cases, seroprevalence was 31.8%. A significant positive direct association existed between seroprevalence and the distances to the Ebola virus-epidemic area. This study suggests that dogs can be infected by Ebola virus and that the putative infection is asymptomatic.