Characterization of Plasmodium Falciparum Resistance Genes to Common Antimalarial Drugs in Semi-urban Areas of Burkina Faso.

INTRODUCTION: Malaria still remains the most frequent parasitic disease on the world with, in 2022, 249 million cases and 608,000 deaths worldwide. Malaria control is compromised by the spread of the parasite's resistance to available antimalarials. The objective of our study is to characterize the Plasmodium falciparum resistance genes to common antimalarial drugs in semi-urban areas of Burkina Faso. MATERIALS AND METHODS: This is a prospective cross-sectional study whose collection took place from June to October 2021 and from June to October 2022 in five health facilities in Burkina Faso. The molecular analysis based on PCR-RFLP took place from January to June 2023 at Centre National de Recherche et de Formation (CNRFP) to determine resistance genes such as Pfcrt, Pfmdr1, Pfdhps, and Pfdhfr. RESULTS: A total of 150 samples were analyzed giving a prevalence of 46.67, 1.33, 0.67, 20, 82, and 4.67%, for Pfcrt 76 T, Pfmdr1 86Y, Pfdhps 437G, Pfdhfr 51I, Pfdhfr 59R, and Pfdhfr 108N mutations, respectively. There are no mutations observed Pfdhps 540E and Pfdhfr 164L positions. However, mutation on Pfdhfr 59R position was the most common. In addition, triple mutation (Pfdhps 437G + Pfdhfr 59R + Pfdhfr 108N) was found with a low frequency which is 0.67%. CONCLUSION: Surveillance of Plasmodium falciparum resistance markers to antimalarial drugs, remains one of the priorities in the context of the control or malaria elimination.

Plasmodium falciparum Genetic Diversity and Resistance Genotype Profile in Infected Placental Samples Collected After Delivery in Ouagadougou.

Purpose: Intermittent preventive treatment with sulfadoxine-pyrimethamine is widely used for the prevention of malaria in pregnant women in Africa. Known resistance cases of sulfadoxine-pyrimethamine during pregnancy need to be follow up to support IPTp implementation in Burkina Faso. However, data on the development and spread of resistance to this molecule are lacking. This study aimed to investigating the genetic diversity of P. falciparum and the mutation prevalence in the dhfr and dhps genes infected from postpartum infected placentas. Patients and Methods: This was a prospective and cross-sectional study conducted between April 2019 and March 2020 in four health districts of Ouagadougou capital city. From the placentas collected after delivery, P. falciparum detection and mps1 and msp2 polymorphism analysis were performed by nested PCR. The resistance profile was checked after analyzing the mutation point on dhfr and dhps genes. Results: PCR-positive samples were estimated at 96% for msp1 and 98% for msp2. The polymorphism analysis showed that the RO33 and 3D7 allelic families were the most widespread with 62.5% and 91.83%, respectively. Multiple infections by msp1 and msp2 were frequent with 12.50% and 92.92%, respectively. The prevalence of individual dhfr mutation point, 51I, 108A, and 59R, was 1.96, 15.68, and 7.84, respectively, and the dhps mutation point, 437G, was 3.92. There is no detected mutation at the point 164L and 540E. The triple (51I+108A+59R) in dhfr and quadruple (51I+108A+59R+ 437G) mutation were not found. Conclusion: The results showed that Plasmodium falciparum has a high genetic diversity of msp1 and msp2. This suggests that dhfr and dhps mutant genotypes are potential early warning factors in the increase in the sulfadoxine-pyrimethamine resistance.

Ethnopharmacological Value and Biological Activities via Antioxidant and Anti-Protein Denaturation Activity of Morinda lucida Benth and Momordica charantia L. Leaves Extracts from Benin.

Momordica charantia Linn. (Cucurbitaceae), the wild variety of bitter melon, and Morinda lucida Benth (Rubiaceae) were commonly used as a popular folk medicine in Benin. This study aimed to appreciate the ethnopharmacological knowledge and evaluate the antioxidant and anti-inflammatory effects of M. charantia and M. lucida leaves extracts. Semi-structured surveys supported by individual interviews were conducted with herbalists and traditional healers in southern Benin. The antioxidant activities were evaluated by a micro-dilution technique using ABTS and FRAP methods. These activities were supported by cyclic voltammetry analysis. The anti-inflammatory activity was evaluated by the albumin denaturation method. The volatile compounds were analysed by GC-MS analysis. All the respondents involved in this study have good knowledge of the two plants. We identify 21 diseases grouped into five categories of condition. The two plants' extracts possess variable antioxidant capacity. Indeed, all the active extracts of M. charantia presented an IC50 < 0.078 mg/mL, while the extracts of M. lucida had an IC50 up to 0.21 ± 0.02 mg/mL. For anti-inflammatory activity, a dose-response activity (p < 0.001) was observed in the protein denaturation inhibition rate of the extracts. It should be noted that the highest inhibition rate (98.34 ± 0.12) of the albumin denaturation was observed with M. lucida dichloromethane extract. A total of 59 volatile compounds were identified by GC-MS analysis in the extracts of the two plants. The M. charantia ethyl acetate extract shows the presence of 30 different compounds with a relative abundance of 98.83%, while that of M. lucida shows 24 compounds with a relative abundance of 98.30%. These plants are potential candidates to discover new compounds with therapeutic properties that could be used to solve public health problems.

Molecular Characterization and the Antimicrobial Resistance Profile of Salmonella spp. Isolated from Ready-to-Eat Foods in Ouagadougou, Burkina Faso.

The emergence of antimicrobial-resistantfood-borne bacteria is a great challenge to public health. This study was conducted to characterize and determine the resistance profile of Salmonella strains isolated from foods including sesames, ready-to-eat (RTE) salads, mango juices, and lettuce in Burkina Faso. One hundred and forty-eight biochemically identified Salmonella isolates were characterized by molecular amplification of Salmonella marker invA and spiC, misL, orfL, and pipD virulence genes. After that, all confirmed strains were examined for susceptibility to sixteen antimicrobials, and PCR amplifications were used to identify the following resistance genes: bla TEM, temA, temB, StrA, aadA, sul1, sul2, tet(A), and tet(B). One hundred and eight isolates were genetically confirmed as Salmonella spp. Virulence genes were observed in 57.4%, 55.6%, 49.1%, and 38% isolates for pipD, SpiC, misL, and orfL, respectively. Isolates have shown moderate resistance to gentamycin (26.8%), ampicillin (22.2%), cefoxitin (19.4%), and nalidixic acid (18.5%). All isolates were sensitive to six antibiotics, including cefotaxime, ceftazidime, aztreonam, imipenem, meropenem, and ciprofloxacin. Among the 66 isolates resistant to at least one antibiotic, 11 (16.7%) were multidrug resistant. The Multiple Antimicrobial Resistance (MAR) index of Salmonella serovars ranged from 0.06 to 0.53. PCR detected 7 resistance genes (tet(A), tet(B), bla TEM, temB, sul1, sul2, and aadA) in drug-resistant isolates. These findings raise serious concerns because ready-to-eat food in Burkina Faso could serve as a reservoir for spreading antimicrobial resistance genes worldwide.

Antimalarial drugs resistance genes of Plasmodium falciparum: a review.

Malaria remains the most common parasitic disease on the planet, with 229 million cases and 409,000 deaths worldwide in 2019, including 274,030 children under the age of 5. It is one of the most important infectious diseases in the world and its control is compromised by the spread of the parasite's resistance to antimalarial drugs. This study aims to review the literature of resistant Plasmodium falciparum genes over the past twenty years. One hundred and five (105) articles were collected and read while the resistance of P. falciparum was being studied. Several P. falciparum gene resistances antimalarial drugs were discovered over the past twenty years. The most recent one is the Kelch13 gene of P. falciparum (Pfkelch13) which has showed resistance to artemisinin in Asia. In Africa, this gene represents a potential candidate for resistance to artemisinin, although no resistance was reported.

Technological and microbiological characteristics of indigenous food produced in Gabon.

The purpose of this study was to provide contextual information on indigenous food's technologies and safety from Gabon. The strategic focus being to promote local food with enhanced nutritional value and improved safety. An investigation and monitoring were carried out to elucidate their process flow diagrams and to identify safety failures. Samples were taken for microbiological analysis using conventional culture-based techniques. Detection and identification of Salmonella in samples were confirmed using PCR based method by targeting invasion plasmid antigen B (IpaB) gene. The investigation shows that women play a protagonist role in the technical know-how of Gabonese indigenous foods in a context that is evolving towards the disappearance of this knowledge. The food production process remains archaic, which makes the environment impact on food safety. Indeed, the proximity of food manufacturing environment to animals, waste, or latrines coupled with the lack of hygiene and manufacturing practices affect the quality of these foods. This is reflected in our study's microbiological results, namely, Aerobic Mesophilic Bacteria ranged from 3.53 to 11.96 log CFU/g and indicators of fecal contaminations of up to 8.21 log CFU/g. Salmonella is detected in 18.69% of samples. The presence of these bacteria is a risk for consumer health. Although some of these foods can be considered as a fermented food, the producers should be further educated and encouraged to take preventive measures to ensure the quality of these food products. A much more subtle approach based on microbial ecology of these foods should be explored for better exploitation.

Exploring Antioxidant and Enzymes (A-Amylase and B-Glucosidase) Inhibitory Activity of Morinda lucida and Momordica charantia Leaves from Benin.

BACKGROUND: Momordica charantia Linn. (Cucurbitaceae), the wild variety of bitter melon and Morinda lucida Benth (Rubiaceae) were commonly used as a popular folk medicine in Benin. This research focused to measure the antioxidant and enzyme inhibitory effects of M. charantia and M. lucida leaves and their antidiabetic activity. METHODS: Antioxidant activities were evaluated by micro-dilution technique using DPPH free radical scavenging activity and ß-carotene-linoleate bleaching assay. The α-amylase inhibition assay was carried out utilizing the 3,5-dinitrosalicylic acid procedure, while ß-glucosidase inhibition assay was demonstrated using as substrate p-nitrophenyl-ß-D-glucopyranoside (PNPG). HPLC-DAD analysis was realized using a high-performance liquid chromatography systems with diode-array detector, L-3000. RESULTS: Chlorogenic acid, epicatechin, daidzein, rutin, naringin, quercetin, naringenin and genistein were identified as polyphenol compounds in the both plants extract. Dichloromethane and ethyl acetate extracts showed a good α-amylase inhibitory activity (56.46 ± 1.96% and 58.76 ± 2.74% respectively). M. lucida methanolic extract has shown IC50 of 0.51 ± 0.01 mg/mL, which is the lowest for DPPH scavenging activity. M. lucida dichloromethane extract showed the highest inhibitory capacity of ß-glucosidase activity (82.11. ± 2.15%). CONCLUSION: These results justify some traditional medicinal uses of both plants. The purified fractions could be used in future formulations, possibly incorporated in functional foods to combat certain diseases.

Assessment of safety and quality of fermented milk of camels, cows, and goats sold and consumed in five localities of Burkina Faso.

BACKGROUND AND AIM: Fermented milk is food produced and consumed all over the world and plays an important role in human nutrition. This work aimed to evaluate the microbiological and physicochemical quality and mineral composition of fermented milk consumed in Burkina Faso. MATERIALS AND METHODS: A total of 114 samples of fermented milk from camels, goats, and cows were purchased in the market in five localities in Burkina Faso; Bobo Dioulasso, Djibo, Dori, Gorom-Gorom, and Sebba. Microbiological and physical parameters were monitored using standards methods. RESULTS: Microbiological analysis of fermented milks showed high average values of 7.60±1.50×109 colony-forming unit per milliliter (CFU/ml), 5.72±3.60×107 CFU/ml, 5.53±2.00×105 CFU/ml, 1.97±0.18×103 CFU/ml, 1.98±0.25×103 CFU/ml, and 0.10±0.09×103 CFU/ml for total microbial flora, lactic acid bacteria, yeasts and molds, Staphylococcus aureus, total coliforms, and thermotolerant coliforms, respectively. None of the samples were contaminated by Salmonella or Shigella. The average values of pH, acidity, dry matter, ash, fats, proteins, and total carbohydrates content of samples were ranged, respectively: 3.830-4.137, 1.888-2.822%, 8.271-13.004%, 0.199-0.476%, 1.210-3.863%, 2.125-3.764%, and 3.080-5.428 % (w/w). Na/K and Ca/Mg ratio ranged from 0.104 to 0.909 and from 3.392 to 16.996, respectively. Total microbial flora, yeasts and molds, total coliforms, fats, calcium, potassium, iron, and zinc were significantly different. CONCLUSION: This research contributed in the evaluation of the hygienic and nutritional qualities of local fermented milk. Results obtained in this study confirm the need to set up the training program on the sanitary condition to traditional maker's to ensure the good fermented milk with high organoleptic and nutritional qualities.

Wastewater used for urban agriculture in West Africa as a reservoir for antibacterial resistance dissemination.

State of art metagenomics were used to investigate the microbial population, antibiotic resistance genes and plasmids of medical interest in wastewater used for urban agriculture in Ouagadougou (Burkina Faso). Wastewater samples were collected from three canals near agricultural fields in three neighbourhoods. Assessment of microbial population diversity revealed different microbial patterns among the different samples. Sequencing reads from the wastewaters revealed different functional specializations of microbial communities, with the predominance of carbohydrates and proteins metabolism functions. Eleven pathogen-specific and 56 orthologous virulence factor genes were detected in the wastewater samples. These virulence factors are usually found in human pathogens that cause gastroenteritis and/or diarrhoea. A wide range of antibiotic resistance genes was identified; 81 are transmissible by mobile genetic elements. These included seven different extended spectrum ß-lactamase genes encoding synthesis of four enzyme families, including two metallo-ß-lactamases (blaAIM-1 and blaGES-21). Ten different incompatibility groups of Enterobacteriaceae plasmid replicons (ColE, FIB, FIC, FII, P, Q, R, U, Y, and A/C), and 30 plasmid replicon types from Gram-positive bacteria. All are implicated in the wide distribution of antibiotic resistance genes. We conclude that wastewater used for urban agriculture in the city represents a high risk for spreading bacteria and antimicrobial resistance among humans and animals.

Safety of ready-to-eat chicken in Burkina Faso: Microbiological quality, antibiotic resistance, and virulence genes in Escherichia coli isolated from chicken samples of Ouagadougou.

In Burkina Faso, flamed/grilled chickens are very popular and well known to consumers. The aim of this study was to evaluate the microbiological quality, the antibiotic resistance, and the virulence gene from Escherichia coli isolated from these chickens in Ouagadougou. A total of 102 grilled, flamed, and fumed chickens were collected in Ouagadougou and analyzed, using standard microbiological methods. All E. coli isolates were checked with the antimicrobial test and also typed by 16-plex PCR. The mean of aerobic mesophilic bacteria (AMB) and thermo-tolerant coliforms (TTC) was found respectively between 6.90 ± 0.12 × 107 CFU/g to 2.76 ± 0.44 × 108 CFU/g and 2.4 ± 0.82 × 107 CFU/g to 1.27 ± 0.9 × 108 CFU/g. E. coli strains were found to 27.45%. Forty samples (38.24%) were unacceptable based on the AMB load. Fifty-nine samples (57.85%) were contaminated with TTCs. Low resistance was observed with antibiotics of betalactamin family. Diarrheagenic E. coli strains were detected in 21.43% of all samples. This study showed that flamed/grilled chickens sold in Ouagadougou could pose health risks for the consumers. Need of hygienic practices or system and good manufacturing practices is necessary to improve the hygienic quality of flamed/grilled chickens. Our results highlight the need of control of good hygiene and production practices to contribute to the improvement of the safety of the products and also to avoid antibiotic resistance. Slaughter, scalding, evisceration, plucking, bleeding, washing, rinsing, preserving, grilling, and selling may be the ways of contamination.

Molecular Characterization of Diarrheagenic Escherichia Coli in Children Less Than 5 Years of Age with Diarrhea in Ouagadougou, Burkina Faso.

Diarrheagenic Escherichia coli (DEC) is important bacteria of children's endemic and epidemic diarrhea worldwide. The aim of this study was to determine the prevalence of DEC isolated from stool samples collected from children with acute diarrhea living in Ouagadougou, Burkina Faso. From August 2013 to October 2015, stool samples were collected from 315 children under 5 years of age suffering from diarrhea in the "Centre Médical avec Antenne Chirurgicale (CMA)" Paul VI and the CMA of Schiphra. E. coli were isolated and identified by standard microbiological methods, and the 16-plex PCR method was used to further characterize them. Four hundred and nineteen (419) E. coli strains were characterized, of which 31 (7.4%) DEC pathotypes were identified and classified in five E. coli pathotypes: 15 enteroaggregative E. coli (EAEC) (48.4%), 8 enteropathogenic E. coli (EPEC) (25.8%) with 4 typical EPEC and 4 atypical EPEC, 4 enteroinvasive E. coli (EIEC) (12.9%), 3 enterohemorrhagic E. coli (EHEC) 9.67%, and 1 enterotoxigenic E. coli (ETEC) 3.2%. The use of multiplex PCR as a routine in clinical laboratory for the detection of DEC would be a useful mean for a rapid management of an acute diarrhea in children.

Characterization of Diarrheagenic Escherichia coli Isolated in Organic Waste Products (Cattle Fecal Matter, Manure and, Slurry) from Cattle's Markets in Ouagadougou, Burkina Faso.

Cattle farming can promote diarrheal disease transmission through waste, effluents or cattle fecal matter. The study aims to characterize the diarrheagenic Escherichia coli (DEC) isolated from cattle feces, manure in the composting process and slurry, collected from four cattle markets in Ouagadougou. A total of 585 samples (340 cattle feces, 200 slurries and 45 manures in the composting process) were collected from the four cattle markets between May 2015 and May 2016. A multiplex Polymerase Chain Reaction (PCR), namely 16-plex PCR, was used to screen simultaneously the virulence genes specific for shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) and enteroaggregative E. coli (EAEC). DEC was detected in 10.76% of samples. ETEC was the most prevalent (9.91%). STEC and EAEC have been observed with the same rate (0.51%). ETEC were detected in 12.64% of cattle feces, in 6.66% of manure in the composting process and in 5% of slurry. STEC were detected in 0.58% of cattle feces and in 2.22% of manure in the composting process. EAEC was detected only in 1% of slurry and in 2.22% of manure in the composting process. ETEC strains were identified based on estIa gene and/or estIb gene and/or elt gene amplification. Of the 58 ETEC, 10.34% contained astA, 17.24% contained elt, 3.44% contained estIa and 79.31% contained estIb. The two positive EAEC strains contained only the aggR gene, and the third was positive only for the pic gene. The results show that effluent from cattle markets could contribute to the spreading of DEC in the environment in Burkina Faso.