Evaluation of sanitary risks associated with the consumption of street food in the city of Yaoundé (Cameroon): case of braised fish from Mvog-Ada, Ngoa Ekélé, Simbock, Ahala and Olézoa.

An investigation was performed to evaluate the braising fish practices and the level of hygiene of the sellers in the city of Yaoundé. Thereafter, five braised carp samples were collected from each of the five selected sites and their microbiological quality assessed. The results showed that all the braising sites do not satisfy the Good Hygiene and Manufacturing Practices. Pathogenic microorganisms such as coliforms (2.17 ± 0.08 to 5.94 ± 0.38 Log cfu/g), E. coli (3.71 ± 0.20 to 6.21 ± 0.48 Log cfu/g) and Salmonella spp. were found in braised carp samples at levels higher than the microbiological criteria of the European Commission which are 1, 1 and 0 Log cfu/g, respectively. Spoilage microorganisms such as yeasts (2.39 ± 0.18 to 3.09 ± 0.23 Log cfu/g) and moulds (2.30 ± 0.10 to 2.92 ± 0.18 Log cfu/g) as well as spore-forming (3.02 ± 0.18 to 4.86 ± 0.35 Log cfu/g) and anaerobic (3.74 ± 0.28 to 4.52 ± 0.38 Log cfu/g) bacteria were also found at higher levels. The study highlights the poor microbiological quality of braised fish sold in the city of Yaoundé.

Pharmacological evidence of Vitex thyrsiflora, Entandrophragma cylindricum, and Anonidium mannii used for the management of inflammation in Cameroon.

Background Inflammation is the most common health problem faced in life relating to a vast number of diseases. The present study evaluated the pharmacological effect of three plants (Vitex thyrsiflora, Entandrophragma cylindricum, and Anonidium mannii) commonly used in the Cameroon pharmacopeia for the management of inflammatory response. Methods The pharmacological effect was characterized by the antioxidant capacity, anti-inflammatory, analgesic, and antipyretic properties of the ethanol extracts of the three plants. Antioxidant capacity was determined using total phenolic content, total flavonoid content, hydrogen peroxide, ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) radical scavenging assays. Anti-inflammatory activity was evaluated in vitro by protein denaturation and hypotonic-induced hemolysis methods and in vivo by carrageenan paw edema method. Analgesic and antipyretic activities were studied in vivo using acetic acid-induced writhing and brewer's yeast-induced hyperpyrexia models. Results All selected extracts showed high phenolic (15.93-64.45 mgCAE/g) and flavonoid (336.03-1053.48 mgCAE/g) contents and high ferric reducing power (288.75-364.91 mgCAE/g). These extracts exhibited good DPPH (IC50 = 0.30-1.65 µg/mL), ABTS (IC50 = 0.52-1.90 µg/mL), and H2O2 (IC50 = 1.40-3.55 µg/mL) radical scavenging activities. All extracts inhibited protein denaturation (6.79-82.27%) and protected the erythrocyte membrane from lysis induced by hypotonic solution (18.90-88.00%). The extracts significantly reduced dose-dependent paw edema (p < 0.05), fever, and abdominal writhing (p < 0.001) especially at 400 mg/kg. Conclusions All extracts exhibited interesting antioxidant properties, as well as significant anti-inflammatory, analgesic, and antipyretic effects.

Effect of fractioning on antibacterial activity of n-butanol fraction from Enantia chlorantha stem bark methanol extract.

BACKGROUND: Enantia chlorantha is a plant belonging to Annonaceae Family. The Barks and leaves are used traditionally to treat infectious diseases. Earlier studies highlighted the antibacterial activity of stem barks methanol extract. This study is thus aimed at investigating the effect of fractionation on antibacterial activity of its n-butanol fraction. METHODS: The extract of E. chlorantha stem barks was obtained by maceration in methanol and then subjected to a liquid/liquid partition by successive depletion with solvents of increasing polarity. The n-butanol fraction was fractionated by adsorption chromatography on silica gel. A product was isolated from the dichloromethane/methanol (2%) fraction and the structure was determined on the basis of spectroscopic data; Proton Nuclear Magnetic Resonance (1H NMR), Carbon-13 Nuclear Magnetic Resonance (13C NMR), Heteronuclear Multiple Bond Correlation (HMBC), H-correlation spectroscopy (H-COSY), attached proton test (APT), heteronuclear multiple quantum coherence (HSQC). The antibacterial activity was evaluated by broth microdilution method against six reference strains and eight clinical bacterial strains. RESULTS: The n-butanol fraction was found to be active with MIC values ranging from 32 to 256 µg/mL. The FA sub-fraction was more efficient among the eight sub-fractions, the n-butanol fraction and comparable to Chloramphenicol used as reference antibiotic. The product obtained was elucidated as palmitin. The antibacterial activity of the latter was comparable to that of Chloramphenicol on one reference strain and 4 of the 6 clinical strains. CONCLUSION: The FA sub-fraction had better antibacterial activity than the n-butanol fraction and other sub-fractions, and possibly palmitin was the active substance responsible for the antibacterial activity of E. chlorantha.

Effect of Fractioning on Antibacterial Activity of Enantia chlorantha Oliver (Annonaceae) Methanol Extract and Mode of Action.

Infectious diseases caused by bacteria constitute the main cause of morbidity and mortality throughout the world and mainly in developing countries. In this work, the influence of fractioning and the mode of action of stem barks methanol extract of Enantia chlorantha were investigated. The aim was to optimize the antibacterial activity of the methanol extract. The extract was prepared by maceration of barks powder in methanol. Fractioning was done using increasing solvents polarity. Standard phytochemical methods were used for phytochemical screening. Minimum Inhibitory Concentrations (MIC) and Minimum Bactericidal Concentration (MBC) of the methanol extract and fractions were determined using broth microdilution method. The studied mode of action of both methanol extract and n-butanol fraction included antibiofilm activity, H+-ATPase-mediated proton pumping assay, salt tolerance, and cells cycle. The methanol extract of E. chlorantha stem barks was found to be active on all the bacteria tested (32 ≤ MIC ≤ 512 µg/mL), its activity being significant (MIC < 100 µg/ml) out of 5 of the 28 clinical isolates used. Salmonella enterica serovar paratyphi A was the most sensitive (32 µg/mL). Compared to the extract and other fractions, the n-butanol fraction was found to be more active (32 ≤ MIC ≤ 256). Significant antibacterial activity of this fraction was observed out of 10 of the 28 bacterial isolates and 3 out of 7 bacterial strains. Lowest MIC values (32 µg/ml) of this fraction were obtained with Escherichia coli (136), Pseudomonas aeruginosa (CIP 76110), and Salmonella enterica serovar typhi 9. The methanol extract of E. chlorantha and its n-butanol fraction revealed several modes of action including the prolongation of the latency phase of the bacterial growth, the inhibition of the pump with protons H+ - ATPases bacterial, the loss of the salt tolerance of the Staphylococcus aureus, and inhibition of the formation of the bacterial biofilm. The present results showed that the n-butanol fraction of the methanol stem barks extract of E. chlorantha possess the essential antibacterial components and could best be used to fight against bacterial infections as compared to methanol extract.

Antibacterial and antioxidant properties of crude extract, fractions and compounds from the stem bark of Polyscias fulva Hiern (Araliaceae).

BACKGROUND: In our previous work, the dichloromethane-methanol (1:1 v/v) extract, fractions and isolated compounds from Polyscias fulva stem bark showed interesting antifungal activity. As a continuity of that work, this study aimed to bring out complementary informations about the antimicrobial properties of P. fulva stem bark that may be useful in the standardization of phytomedicine from this plant. METHODS: The antibacterial activities of the crude extract, fractions (n-hexane, ethyl acetate, n-butanol and residual) and isolated compounds from Polyscias fulva stem bark were assayed by broth microdilution techniques. Their antioxidant activity were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), pyrogallol (superoxide anion) and ß-carotene - linoleic acid assays. RESULTS: The crude extract presented antibacterial activities against S. typhi (ATCC 6539), E. aerogenes (ATCC 13045), P. aeruginosa (PA01) and E. coli (ATCC 10536) with MIC values of 2000 to 8000 µg/ml. The fractionation led the ethyle acetate and n-butanol fractions relatively more active (MIC = 500 to 1000 µg/ml) as compared to the crude extract. ß-sitosterol and 3-O-α-L- arabinopyranosyl-hederagenin were the most active compounds on the tested bacteria with MIC values ranging from 6.25 to 100 µg/ml. The most sensitive was P. aeruginosa (PA01) on which all the tested compounds were active with MICs ranging from 6.25 to 400 µg/ml. Among all the tested substances, the crude extract (RSa50 = 84.86 µg/ml) and the methyl atrarate (RSa50 = 14.77 µg/ml), showed the highest scavenging activities against DPPH free radicals and those arising from the oxidation of the linoleic acid respectively. CONCLUSION: From this study, the results obtained reveal that the stem bark of P. fulva possesses antibacterial and antioxidant activities. It may then be useful in the development of an antimicrobial phytomedicine with a large spectrum of actvity endowed with antioxidant properties which can be standardised based on the isolated compounds.

Influence of Processing Methods on Proximate Composition and Dieting of Two Amaranthus Species from West Cameroon.

The effects of various processing methods on the proximate composition and dieting of Amaranthus hybridus and Amaranthus cruentus from West Cameroon were investigated in this study. Both amaranths leaves were subjected to same treatments (sun-dried and unsliced, sliced and cooked), milled, and analysed for their mineral and proximate composition. Thirty-Six Wistar albino rats of 21 to 24 days old were distributed in six groups and fed for 14 days with 10% protein based diets named D0 (protein-free diet), DI (egg white as reference protein), DII (sun-dried and unsliced A. hybridus), DIII (cooked and sliced A. hybridus), DIV (sun-dried and unsliced A. cruentus), and DV (cooked and sliced A. cruentus). The protein bioavailability and haematological and biochemical parameters were assessed in rats. The results showed that K, P, Mg, Zn, and Fe had the higher content in both samples regardless of processing method. The sun-dried and unsliced A. cruentus contained the highest value of crude protein 32.22 g/100 g DM (dry matter) while the highest crude lipid, 3.80 and 2.58%, was observed, respectively, in sun-dried and unsliced A. hybridus and cooked and sliced A. cruentus. Cooked and sliced A. hybridus and A. cruentus contained high crude fiber of 14 and 12.18%, respectively. Rats fed with diet DIII revealed the best protein bioavailability and haematological parameters whereas 100% mortality rate was recorded with group fed with diet DIV. From this study, it is evident that cooked and sliced A. hybridus and A. cruentus could play a role in weight reduction regimes.

Anti-Staphylococcus aureus activity of methanol extracts of 12 plants used in Cameroonian folk medicine.

BACKGROUND: The emergence of bacterial infections including those associated with Staphylococcus aureus causes a benefit of interest to medicinal plants as an effective means of control. The present study was designed to investigate the activities of 12 selected Cameroonian medicinal plants against S. aureus isolates. METHODS: The plant extracts were prepared by maceration in methanol at laboratory temperature. Qualitative phytochemical analysis was performed by chemical reaction methods. The broth microdilution method was used to evaluate the activities of plant extracts against 11 S. aureus clinical isolates. RESULTS: Dacryodes edulis was found to have significant antibacterial activity on all the S. aureus isolates (MIC = 64-256 µg/ml). Occimum gratissimum revealed significant inhibitory activity on 9 of the 11 isolates while Commelina erecta and Spilanthes filicaulis revealed similar results on 6 of the 11 clinical isolates. CONCLUSION: The present findings showed that D. eduli, O. gratissimum, C. erecta and S. filicaulis possess interesting inhibitory properties against S. aureus species. These plants could therefore be good candidates to overcome infectious diseases associated with these microorganisms.

Immunomodulatory activity of methanol leaf extracts of Cameroonian medicinal plants.

BACKGROUND: Medicinal plants have been used for centuries and have become part of complementary medicine worldwide because of their health benefits. Some have been successfully used directly in the treatment and prevention of infectious diseases or indirectly by stimulating the immune system. In the present study, the immunomodulatory activity of the methanol extracts of Adenocarpus mannii, Caucalis melanantha, Ocimum gratissimum, Asystasia intrusa and Clematis chinensis leaves was investigated. METHODS: The extracts were prepared by maceration of dry leaves' powder in methanol. Phytochemical analysis was carried out by chemical reaction methods. The activity of plant extracts was evaluated in in vitro cell cultures by measuring their effect on nitric oxide production by peritoneal macrophages, the proliferation of lymphocytes and the cytotoxic effect on macrophages. The A. mannii extract was further evaluated at 50 and 100 mg/kg body weight in mice for the stimulation of delayed-type hypersensitivity (DTH) reactions and the ability to reverse the myelosuppression induced by cyclophosphamide. RESULTS: All the extract inhibited nitric oxide production by peritoneal macrophages, the highest activity was achieved with C. chinensis extract. However, these extracts did not significantly affect the viability of macrophages. A. mannii, A. intrusa and C. chinensis extracts exhibited stimulatory activity on peripheral blood lymphocytes, whereas C. melanantha and O. gratissimum extracts displayed inhibitory activity. In vivo, the A. mannii extract significantly increased the DTH reaction in mice from 50 mg/kg. This extract also showed a significant increase in the white blood cells and relative weight of the spleen and liver. CONCLUSIONS: These results suggest that the A. mannii, C. melanantha, O. gratissimum, A. intrusa and C. chinensis methanol extracts possess immunomodulatory activity. This constitutes additional data on the well-known biological properties of these plants.

Toxicological studies of stem bark extract from Schefflera barteri Harms (Araliaceae).

BACKGROUND: The use of herbal medicines as complements or alternatives to orthodox medicines has been on the increase. There has been the erroneous belief that these medicines are free from adverse effects. Schefflera barteri is popularly used in the West region of Cameroon for the treatment of various diseases such as diarrhea, spasm, pneumonia and animals bite. Considering the ethnopharmacological relevance of this plant, this study was designed to investigate the possible toxic effects of the stem bark extract of S. barteri. METHODS: The extract was prepared by maceration of stem bark dry powder in methylene chloride/methanol mixture. Phytochemical analysis was performed by chemical reaction method. Oral acute toxicity study was carried out by administering single geometric increasing doses (2 to 16 g/kg body weight) of plant extract to Swiss albino mice. For sub-acute toxicity study, repeated doses (100, 200, 400 and 800 mg/kg bw) of plant extract were given to Wistar albino rats for 28 consecutive days by oral route. At the end of the treatment period, hematological and biochemical parameters were assessed, as well as histopathological studies. RESULTS: Phytochemical analysis of stem bark extract of S. barteri revealed the presence of anthocyanins, anthraquinons and saponins. Acute toxicity results showed that the LD50 was greater than 16000 mg/kg. Sub-acute treatment significantly (P < 0.05) increased the level of serum transaminase, proteins and HDL cholesterol. On the other hand, the extract significantly (P < 0.05) reduced the level of leucocytes as well as neutrophils, basophils and monocytes in female. No significant variation of serum creatinine, LDL cholesterol, serum triglycerides as well as liver, spleen, testicles and ovaries proteins was noted. Histopathological analysis of organs showed vascular congestion, inflammation of peri-portal and vacuolization of hepatocytes at the level of the liver. Leucocytes infiltration of peri-portal veins were noticed on lungs and liver cells as well as inflammatory peri-bronchial and basal membranes seminar tube merely joined on lungs and testis respectively. CONCLUSION: The results suggest that acute administration of the stem bark extract of S. barteri is associated with signs of toxicity, administration over a long duration provokes hepatotoxicity, testes and lungs toxicities.

Antifungal properties of a new terpernoid saponin and other compounds from the stem bark of Polyscias fulva Hiern (Araliaceae).

BACKGROUND: In our previous studies, it was evident that the dichloromethane-methanol (1:1 v/v) stem barks extract of Polyscias fulva and fractions (ethyl acetate, n-butanol and residue) demonstrated interesting antidermatophytic activities. So, as a continuity of that, this work aimed at identifying active principles with antifungal properties from P. fulva that could be used as markers for possible standardization of this plant as phytomedicine. METHODS: The ethyl acetate, n-butanol and residual fractions of the dichloromethane-methanol (1:1 v/v) stem bark extract of Polyscias fulva were further fractionated by column chromatography and the structures of isolated compounds elucidated based on their spectroscopic data in comparison with existing literature information. Antifungal activity was assayed by broth microdilution techniques on yeasts and dermatophytes spores. RESULTS: The fractionation of the crude dichloromethane-methanol (1:1 v/v) stem bark extract of Polyscias fulva led to the isolation of 10 known compounds (1 to 10) and one new saponin (11: 3-O-[α-L-rhamnopyranosyl (1-2)-α-L-arabinopyranosyl]-28-O-[α-L-4-O-acetyl-rhamnopyranosyl (1-4)-ß-D-glucopyranosyl-(1-6)-ß-D-glucopyranosyl]-hederagenin). Among these compounds, 3-O-α-L- arabinopyranosyl-hederagenin and 3-O-[α-L-rhamnopyranosyl (1-2)-α-L-arabinopyranosyl]-hederagenin were the most active on the tested fungi with MIC values ranging from 0.78 to 100 µg/ml against both yeasts and dermatophytes. CONCLUSION: The results of this work constitute a step forward in the possible development of an antidermatophytic phytomedicine from Polyscias fulva stem bark, the isolated compounds being possible markers for the standardisation.

Antifungal and antioxidant activity of Crassocephalum bauchiense (Hutch.) Milne-Redh ethyl acetate extract and fractions (Asteraceae).

BACKGROUND: Crassocephalum bauchiense is a flowering plant, found in the West Region of Cameroon. Previous studied has highlighted the antibacterial and the dermal toxicological safety as well as the immunomodulatory activities of the ethyl acetate extract of its dry leaves. As an extension of the previous researches, the current work has been undertaken to evaluate the in vitro antifungal and antioxidant activities of C. bauchiense dried leaves ethyl acetate extract and fractions. METHODS: The extract was obtained by maceration in ethyl acetate and further fractionated into six fractions labeled F1 to F6 by flash chromatography. The antifungal activity of the extract and fractions against yeasts and dermatophytes was evaluated using broth microdilution method. Antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO) and ß-carotene - linoleic acid assays. RESULTS: The extract (MIC = 0.125 - 4 mg/ml) was found to be more active on dermatophytes and yeasts compared to the fractions. The ethyl acetate extract and fractions exhibited strong scavenging activity on DPPH (CI50 = 28.57 - 389.38 µg/ml). The fractions F3 and F6 expressed best antioxidant activity on DPPH radicals compared to the crude extract. CONCLUSION: The results of these findings clearly showed that C. bauchiense ethyl acetate extract has a significant antifungal and antioxidant activity. It is therefore a source of active compounds that might be used as antifungal and antioxidant agents.

In vitro and in vivo antidermatophytic activity of the dichloromethane-methanol (1:1 v/v) extract from the stem bark of Polyscias fulva Hiern (Araliaceae).

BACKGROUND: During the last decades, the number of people suffering from dermatophytoses has seriously increased, mainly due to the development of resistant strains of microorganisms to a range of formally efficient antibiotics. Polyscias fulva, a medium size tree which grows in the West Region of Cameroon is traditionally used for local application against dermatoses and orally against venereal infections. The dichloromethane-methanol (1:1 v/v) extract from the stem bark of Polyscias fulva was evaluated for its in vitro and in vivo antifungal activities. METHODS: The plant extract was prepared by maceration of its stem bark powder in CH(2)Cl(2)-MeOH (1:1 v/v). The extract obtained was successively partitioned in hexane, ethyl acetate and n-butanol. Phytochemical screening was performed using standard methods. In vitro antidermatophytic activity was assayed by the well diffusion and broth microdilution methods. The degree of dermal irritation of the crude extract was determined in guinea pigs using the occluded dermal irritation test method. The in vivo antidermatophytic activity of the extract-oil formulation (1.25, 2.5 and 5% w/w concentrations) was evaluated using Trichophyton mentagrophytes-induced dermatophytosis in a guinea pigs model. RESULTS: Phytochemical screening indicated that, the crude extract, ethyl acetate, n-butanol and residue fractions contain in general saponins, tannins, alkaloids, anthraquinones and phenols while the hexane fraction contains only alkaloids. The ethyl-acetate, n-butanol and residue fractions displayed higher antifungal activities (MIC = 0.125-0.5 mg.mL(-1)) against eight dermatophytes as compared to the crude extract (MIC = 0.5-1 mg.mL(-1)). This latter appeared to have slight perceptible erythema effects on guinea pigs as the primary irritation index (PII) was calculated to be 0.54. In vivo, the antidermatophytic activities of the extract-oil formulations were dose-dependent. Griseofulvin-oil 5% at 0.01 g/kg and formulated extract-oil (5%) at 0.1 g/kg eradicated the microbial infection after thirteen and fourteen days of daily treatment respectively. CONCLUSIONS: The results of preclinical in vitro and in vivo evaluations indicate that the extract-oil formulation at 5% may constitute an alternative means to alleviate fungal infections caused by dermatophytes.

Antibacterial and Antioxidant Properties of the Methanolic Extract of the Stem Bark of Pteleopsis hylodendron (Combretaceae).

Pteleopsis hylodendron (Combretaceae) is used in Cameroon and West Africa folk medicine for the treatment of various microbial infections (measles, chickenpox, and sexually transmitted diseases). The antibacterial properties of the methanolic extract and fractions from stem bark of Pteleopsis hylodendron were tested against three Gram-positive bacteria and eight Gram-negative bacteria using Agar-well diffusion and Broth microdilution methods. Antioxidant activities of the crude extract and fractions were investigated by DPPH radical scavenging activity and ß-carotene-linoleic acid assays. The methanolic extract and some fractions exhibited antibacterial activities that varied between the bacterial species (ID = 0.00-25.00 mm; MIC = 781-12500 µg/mL and 0.24-1000 µg/mL). The activity of the crude extract is, however, very weak compared to the reference antibiotics (MIC = 0.125-128 µg/mL). Two fractions (F(E) and F(F)) showed significant activity (MIC = 0.97 µg/mL) while S. aureus ATCC 25922 was almost resistant to all the tested fractions. In addition, the crude extract and some fractions showed good antioxidant potential with inhibition values ranging from 17.53 to 98.79%. These results provide promising baseline information for the potential use of this plant as well as some of the fractions in the treatment of infectious diseases and oxidative stress.