Antioxidant, Anti-Inflammatory, and Anti-Cancer Properties of Amygdalin Extracted from Three Cassava Varieties Cultivated in Benin.

Given that cancer is a disease that is rampant in the world and especially in Africa, where the population has enormous difficulty treating it, plants are a safer and less expensive alternative. Cassava is a plant species valued in Benin because of its numerous medicinal and nutritional virtues. This study evaluated the biological activities of amygdalin from the organs of three cassava varieties most commonly produced in Benin (BEN, RB, and MJ). HPLC analysis was used to quantify amygdalin in cassava organs and derivatives. Phytochemical screening was performed to determine secondary metabolite groups. DPPH and FRAP methods were used to assess antioxidant activity. Cytotoxicity of the extracts was tested on Artemia salina larvae. The anti-inflammatory activity was evaluated in vivo in an albino mouse paw edema model induced by 5% formalin. The anticancer activity was evaluated in vivo on Wistar rats rendered cancerous by 1,2-dimethylhydrazine (DMH) using 5-fluorouracil as a reference molecule. The results showed that the organs of all three-cassava varieties contained glycosides, flavonoids, saponosides, steroids, tannins, coumarins, and cyanogenic derivatives. Young stems and fresh cassava leaves had the highest amygdalin concentrations, with 11,142.99 µg 10 g-1 and 9251.14 µg 10 g-1, respectively. The Agbeli derivative was more concentrated in amygdalin, with a content of 401.56 µg 10 g-1 than the other derivatives. The antioxidant activity results showed that the amygdalin extracts were DPPH radical scavengers with IC50 values ranging from 0.18 mg mL-1 to 2.35 mg mL-1. The cytotoxicity test showed no toxicity of the extracts toward shrimp larvae. Administration of amygdalin extracts from the leaves of BEN and MJ varieties prevents inflammatory edema. The percentages of edema inhibition varied between 21.77% and 27.89%. These values are similar (p > 0.05) to those of acetylsalicylic acid (25.20%). Amygdalin extract of the BEN variety significantly (p < 0.0001) reduces edema. Both BEN extracts inhibited cancer induction with DMH. In preventive and curative treatments, rats fed with amygdalin extracts showed low anti-cancer activity under the effect of DMH and a significant difference in biochemical results. Thus, the organs of all three cassava varieties studied have secondary metabolites and good antioxidant activity. The leaves contain high levels of amygdalin and can be used as anti-inflammatory and anticancer agents.

Resistance and Biofilm Production Profile of Potential Isolated from Kpètè-Kpètè Used to Produce Traditional Fermented Beer.

This study aimed to characterize the pathogenicity of bacteria isolated from the starter of two traditional beers produced and consumed in Benin. After standard microbial identification, species were identified by specific biochemical tests such as catalase, coagulase, and API 20 E. Antibiotic sensitivity was tested according to the French Society of Microbiology Antibiogram Committee. The crystal violet microplate technique evaluated the biofilm production and conventional PCR was used to identify genes encoding virulence and macrolide resistance. According to our data, the traditional starter known as kpètè-kpètè that is used to produce beer is contaminated by Enterobacteriaceae and staphylococci species. Thus, 28.43% of the isolated bacteria were coagulase-negative staphylococci (CNS), and 10.93% coagulase-positive staphylococci (CPS). Six species such as Klebsiella terrigena (1.38%), Enterobacter aerogens (4.14%), Providencia rettgeri (5.51%), Chryseomonas luteola (6.89%), Serratia rubidae (15.16%), and Enterobacter cloacae (27.56%) were identified among Enterobacteriaceae. Those bacterial strains are multi-resistant to conventional antibiotics. The hight capability of produced biofilms was recorded with Enterobacter aerogens, Klebsiella terrigena (100%), Providencia rettgeri (75%), and Staphylococcus spp (60%). Enterobacter cloacae (4%) and coagulase-negative Staphylococcus (5.55%) harbor the macrolide resistance gene. For other strains, these genes were not detected. Foods contaminated with bacteria resistant to antibiotics and carrying a virulence gene could constitute a potential public health problem. There is a need to increase awareness campaigns on hygiene rules in preparing and selling these traditional beers.

Antibiotic Resistance Profiling of Pathogenic Staphylococcus Species from Urinary Tract Infection Patients in Benin.

Staphylococci can cause urinary tract infections (UTIs). These UTIs are among the significant causes of antibiotic resistance and the spread of antibiotic-resistant diseases. The current study is aimed at establishing a resistance profile and determining the pathogenicity of Staphylococcus strains isolated from UTI samples collected in Benin. For this purpose, urine samples (one hundred and seventy) that were collected from clinics and hospitals showed UTI in patients admitted/visited in Benin. The biochemical assay method was used to identify Staphylococcus spp., and the disk diffusion method tested the antimicrobial susceptibility. The biofilm formation ability of the isolates of Staphylococcus spp. was investigated by the colorimetric method. The presence of mecA, edinB, edinC, cna, bbp, and ebp genes was examined by multiplex polymerase chain reaction (PCR). The results showed that Staphylococcus species were identified in 15.29% of all infected individuals and that 58% of these strains formed biofilms. Most Staphylococcus strains (80.76%) were isolated in female samples, and the age group below 30 years appeared to be the most affected, with a rate of 50%. All Staphylococcus strains isolated were 100% resistant to penicillin and oxacillin. The lowest resistance rates were seen with ciprofloxacin (30.8%), gentamicin, and amikacin (26.90%). Amikacin was the best antibiotic against Staphylococcus strains isolated from UTIs. The isolates carried mecA (42.31%), bbp (19.23%), and ebp (26.92%) genes in varying proportions. This study provides new information on the risks posed to the population by the overuse of antibiotics. In addition, it will play an essential role in restoring people's public health and controlling the spread of antibiotic resistance in urinary tract infections in Benin.

Susceptibility and Virulence of Enterobacteriaceae Isolated from Urinary Tract Infections in Benin.

Enterobacteriaceae represent one of the main families of Gram-negative bacilli responsible for serious urinary tract infections (UTIs). The present study aimed to define the resistance profile and the virulence of Enterobacteriaceae strains isolated in urinary tract infections in Benin. A total of 390 urine samples were collected from patients with UTIs, and Enterobacteriaceae strains were isolated according to standard microbiology methods. The API 20E gallery was used for biochemical identification. All the isolated strains were subjected to antimicrobial susceptibility testing using the disc diffusion method. Extended-spectrum beta-lactamase (ESBL) production was investigated using a double-disc synergy test (DDST), and biofilm production was quantified using the microplate method. Multiplex PCR was used to detect uro-virulence genes, namely: PapG, IronB, Sfa, iucD, Hly, FocG, Sat, FyuA and Cnf, using commercially designed primers. More than 26% (103/390) of our samples were contaminated by Enterobacteriaceae strains at different levels. Thus, E. coli (31.07%, 32/103), Serratia marcescens (11.65%, 12/103), Klebsiella ornithinolytica (8.74%, 9/103), Serratia fonticola (7.77%, 8/103) and Enterobacter cloacae (6.80%, 7/103) were identified. Among the isolated strains, 39.81% (41/103) were biofilm-forming, while 5.83% (6/103) were ESBL-producing. Isolates were most resistant to erythromycin, cefixime, ceftriaxone and ampicillin (≥90%) followed by ciprofloxacin, gentamycin, doxycycline and levofloxacin (≥50%), and least resistant to imipenem (27.18%). In regard to virulence genes, Sfa was the most detected (28.15%), followed by IronB (22.23%), iucD (21.36%), Cnf (15.53%), PapG (9.71%), FocG (8.74%), Sat (6.79%), FyuA (5.82%) and Hyl (2.91%). These data may help improve the diagnosis of uropathogenic strains of Enterobacteriaceae, but also in designing effective strategies and measures for the prevention and management of severe, recurrent, or complicated urinary tract infections in Benin.

In vivo anticoccidial activity and immune response of Moringa oleifera and Vernonia amygdalina leaves against Eimeria magna and Eimeria media in rabbits.

Introduction: The aims of this study were to determine the immune response and the anticoccidial activity induced by Moringa oleifera and Vernonia amygdalina leaves in rabbits infected with Eimeria magna and Eimeria media. Methods: Thirty-five-day-old rabbits, free from coccidia, were infested with 2.103 oocysts of Eimeria magna and Eimeria media, then received the acetone extract of the leaves of Moringa oleifera and Vernonia amygdalina at different doses by oral gavage. Results and discussion: The inhibition of the excretion of oocysts was evaluated by the McMaster technique and the levels of cytokines (IL-4 and IL-12) and immunoglobulin IgG were assayed by the ELISA method. The in vivo efficacy on E. magna and E. media oocysts was 95.43% and 96.53% for Moringa oleifera and Vernonia amygdalina at 1000 mg/kg bw against 98% for the positive control. Interestingly the plant extracts increased the production of interleukin (IL) and immunoglobulins (Ig) compared to controls. Plasma IL-4 levels (pg/ml) in rabbits were 128.94 and 131.38; those of IL-12 (pg/ml) were 395.55 and 426.56, and then for those of IgG (µg/ml) were 14.70 and 13.94 respectively with the acetone extracts of Moringa oleifera and Vernonia amygdalina on D14 PT at 1000 mg/kg bw. This study indicates that Moringa oleifera and Vernonia amygdalina can be used as an alternative to synthetic anticoccidials. These plants could be used to increase the resistance of the immune system of rabbits to infestations of Eimeria species in rabbit farms.

Draft Genome Sequences of Four Strains of Plant Growth-Promoting Bacteria Associated with Maize Rhizosphere.

This study presents the draft genome sequences of four strains of rhizobacteria, namely, Bacillus cereus ADO11, Stenotrophomonas maltophilia NAA11, Acinetobacter pittii LAM11, and Serratia marcescens NSA15, which were isolated from maize soils and have the ability to stimulate plant growth. The genome assembly sizes for the strains were 4,476,462 bp, 4,731,402 bp, 4,080,875 bp and 4,959,744 bp, respectively.

Bio-production of lactic and lactobionic acids using whey from the production of cow's milk Wagashi cheese in Benin.

Traditional cheese is the main milk derivative in Bénin. This traditional process is not efficient and generate a lot of whey which has no real use until now. It is just disposed without being environmentally treated. Its use as a source for lactobionic and lactic acids production by Pseudomonas taetrolens and Lactobacillus casei is studied in this work, being also a proposal that can greatly boost economically the dairy sector in the country and reduce the end-of-cycle impact of the residue. To our knowledge, no data is available in the metabolization of Bénin's traditional cheese whey and its potential transformation into commercially valuable products such as lactobionic and lactic acids. With bulk filtration, non-controlled pH batch fermentations and without nutrients supplementation, 66 and 22% of lactose in the traditional cheese whey have been converted into lactobionic acid and lactic acid using Pseudomonas taetrolens and Lactobacillus casei, respectively. Those are important results that encourage to enhance the bioprocesses used in a cost-effective way in order to scale up an industrial production.

Characteristics of Escherichia coli Isolated from Intestinal Microbiota Children of 0-5 Years Old in the Commune of Abomey-Calavi.

Escherichia coli is a commensal bacterium and one of the first bacteria to colonize the digestive tract of newborns after birth. It is characterized by great versatility and metabolic flexibility that allows its survival in different niches. The present study aims at analyzing the diversity of E. coli strains isolated from the intestinal microbiota of children aged from 0 to 5 years in the commune of Abomey-Calavi in Benin. For this purpose, a descriptive and analytical cross-sectional study was conducted. A total of 135 stool samples were collected from the pediatric clinic of Abomey-Calavi. Microbiological analyses were performed according to standard microbiology analytical techniques. The molecular characterization of E. coli was performed by investigating eight genes (dinB, icdA, pabB, polB, putP, trpA, trpB, and uidA) using the PCR technique. The results showed that the average loading rate on stool samples was 3.74 × 107 CFU/g for TAMF. A total of 7 species of bacteria were identified at different proportions: Staphylococcus spp (55.36%), E. coli (14.29%), Klebsiella ornithinolytica (12.5%), Serratia odorifera (5.36%), and Enterobacter aerogenes (5.36%). Interestingly, isolated E. coli presented a resistance of 100% to cefotaxime and aztreonam. In addition, resistances of 95.24% and 50% were observed against erythromycin and nalidixic acid, respectively. The molecular characterization of the isolated E. coli strains allowed us to discover another molecular variation within the isolated strains. Genes encoding the enzymes isocitrate dehydrogenase (icd) and DNA polymerase II (polB) were detected at 96.30% in the isolated E. coli strains. Moreover, the genes encoding the enzymes beta-D-glucuronidase (uidA) and DNA polymerase (dinB) were detected at 88.89% in the isolated E. coli strains. Interestingly, 81.48%, 85.19, 92.59%, and 100% of isolated E. coli strains expressed the genes encoding the enzymes tryptophan synthase subunit A (trpA), proline permease (putP), p-aminobenzoate synthase, and tryptophan synthase subunit B (trpB), respectively. The diversity of E. coli strains reflects the importance of regulatory mechanisms in the adaptation of bacteria to the gut microbiota.

Effect of the application or coating of PGPR-based biostimulant on the growth, yield and nutritional status of maize in Benin.

Biotechnology proposes various ecological approaches to control climatic constraints, soil fertility and plant nutrition using biological products, such as biostimulants to achieve a healthy and environment-friendly agriculture. The aim of this study was to compare the effect of biostimulant-coated maize seed and biostimulant application on the growth, yield and nutritional status of maize in Benin. The trials were set up with 100 producers spread over the whole of Benin. The experimental design was a block of three treatments with 11 replicates per Research-Development (R-D) sites. The maize varieties 2000 SYNEE-W BENIN and TZL COMP 4-W BENIN were used. The best growth (height, stem diameter and leaf area) and yield performances (thousand grains weight and grains yield) were obtained by treatments T2 (Application of biostimulant + ½ NPK-Urea) and T3 (Seed coating with biostimulant + ½ NPK-Urea) compared to the farmers' practice (T1). A significant difference was observed between the different treatments for height, leaf area, 1000 grains weight and maize-grain yield. From one Research-Development site to another, a significant difference was also observed for all parameters. The treatment- Research-Development site interaction was also significant in most areas. The applied or coated biostimulant improved the uptake of nitrogen, phosphorus and especially potassium with higher significant difference compared to the recommended dose of mineral fertilizer. The two techniques of using the biostimulant combined with the half-dose of mineral fertilizer gave the better growth, yield and nutritional status compared to the farmers' practice in all areas study. This biostimulant can be used to ensure food security and sustainable agriculture in Benin.

Phytochemical composition and in vitro biological activities of Morinda citrifolia fruit juice.

Morinda citrifolia is a plant with broad nutraceutical and therapeutic effects and used in the traditional treatment of several ailments. The objective of this work is to investigate the phytochemistry of the fruit juice of M. citrifolia on one hand and on other hand to evaluate its antiradical and antibacterial activity. The phytochemical investigation was carried out by tube staining tests of the extract of two types of fruit juice of M. citrifolia. The antioxidant activity of these juices was evaluated by reducing the DPPH radical method. Concerning the antibacterial activity, it was tested on the in vitro growth of 10 reference bacterial strains using the well diffusion method. Qualitative phytochemistry of M. citrifolia fruit juices revealed the presence of large groups of secondary metabolites including polyphenols, reducing compounds, mucilage and terpernoids. The antioxidant activity of M. citrifolia fruit juices is dose-dependent and higher than that of ascorbic acid. Antimicrobial activity on other hand revealed that fruit juices inhibit growth inhibitory activity of Staphylococcus aureus, Pseudomonas aeruginosa, Proteus mirabilis, S. epidermidis, Proteus vulgaris, Streptococcus oralis, Enterococcus faecalis and Escherichia coli. This observed difference is significant for each juices on the strains (p < 0.001). These results support the use of M. citrifolia in traditional medicine and are the starting points for the development of a new drug to combat both dietary conditions and chronic conditions associated with oxidative stress.