Insights into low biological activity of wax apple (Syzygium samarangense) juice by in vitro phytochemical investigation with special reference to metabolomics

Majumder S, Acharyya S, Ghosh A, Chakraborty S, Sarkar S, Saha S, Bhattacharya M. 2021. Insights into low biological activity of wax apple (Syzygium samarangense) juice by in vitro phytochemical investigation with special reference to metabolomics. Biofarmasi J Nat Prod Biochem 19: 30-38. Wax-apple (Syzygium samarangense) is an Asian waxy and juicy fruit which is reported as antidiarrheal and useful in diseases like dysentery. Moreover, no stronger medicinal evidences have been found associated with this fruit as well as its juice to ensure more biological activities and composition. Being edible, waxy and juicy this fruit is quite popular in coastal areas and islands but, absence of stronger medicinal and biochemical evidences associated with this fruit or its juice has created an worldwide underutilizing status on it. In this research, we intended to investigate phytochemical characteristics and composition of this fruit juice through detailed biochemical tests and metabolomics. Qualitative detection tests for bioactive groups of molecules (tannin, coumarin, cardiac glycosides, terpenoids, flavonoids, phenol etc.), antioxidant assay and antibacterial test simply showed its low in vitro biological activity. A GC-MS based metabolomics was performed where presence of wax components like long chain hydrocarbons have been revealed. Based on the GC-MS based metabolomics a proposed pathway of wax apple’s wax biosynthesis has been established. The overall study not only strongly clarified the absence of bioactive components in this fruit but also proved that the fruit, named wax apple, is actually a source of natural waxes which can be responsible for its claimed antidiarrheal property.

Antifungal activities of methanol extracts of some medicinal plants against germination and growth of Colletotrichum destructivum O’Gara in culture

Authors. 2021. Antifungal activities of methanol extracts of some medicinal plants against germination and growth of Colletotrichum destructivum O’Gara in culture. Asian J Nat Prod Biochem 19: 25-29. Cowpea is an important grain and leafy vegetable in many tropical locations. Anthracnose caused by Colletotrichum destructivum O’Gara is one of the several factors constraining the economic production of the crop in warm humid areas. Use of resistant varieties and synthetic pesticides for control of the disease in the crop are disadvantaged for variability of the pathogen, leading to resistance failure of cultivars in many cowpea growing areas; and pesticide residues in the treated crops which ultimately engender mammalian toxicity respectively. This study evaluated the effects of Alchornea cordifolia, Tabernaemontana pachysiphon and Lantana camara as eco-friendly fungicides against the pathogen in culture. The results showed that the extracts inhibited the fungus in a dose-wise manner. At 75 and100 % concentrations, L. camara strongly inhibited the spore germination and radial growth of the fungus better than all concentrations of T. pachysiphon and A. cordifolia; whereas A. cordifolia extracts exhibited the lowest inhibition at all concentrations. Generally, the fungitoxicity of benomyl was found higher which inhibited 90.15 % spore germination and 96.32 % radial growth of the pathogen. However, at 75%, 100% L. camara and 100% T. pachysiphon which significantly (P?0.05) inhibited (80.02%, 84.21 % and 80.91%) spore germination and (87.33%, 90.87 % and 85. %) radial growth of the fungus respectively compared well with inhibition effects recorded from benomyl. Therefore these plants can be used as viable protectants of cowpea against anthracnose (C. destructivum) in small holder farming systems of the humid tropics; to enhance its production and farm economy.

Short Communication: Analysis of purity and concentration of extracted DNA on salted fish processed food products

Sophian A. 2021. Short Communication: Analysis of purity and concentration of extracted DNA on salted fish processed food products. Asian J Nat Prod Biochem 19: 21-24. Analysis of purity and concentration of extracted DNA on salted fish processed food products was carried out in the microbiology and molecular biology testing laboratory of the Food and Drug Administration in Gorontalo. The purpose of this study was to analyze the extracted DNA on salted fish processed food products based on the concentration and purity values in the A260 / A230 and A260 / A280 wavelengths. The method used for purity and concentration analysis was the absorbance method using a nanophotometer. The samples used were 10 types of salted fish processed food products sampled from 5 (five) traditional markets in Gorontalo City. The sample was extracted using the spin column method with the Dneasy Mericon Food Kit (50) paint kit. 69514 (Qiagen). The research data showed that extracted sample concentration was in the range of 24,600 - 27,150 with an average of 25,745, while purity value measured at A260 / A280 wavelength was obtained with a purity range between 1,668 - 1,768 with an average of 1,729. Based on the results of this study, it can be concluded that the results of DNA extraction carried out on salted fish processed food products show a value that is in the category of good DNA extraction results.

Short Communication: Detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products using Real-Time PCR Multiplex Method

Sophian A, Purwaningsih R, Igirisa RPJ, Amirullah ML, Lukita BL, Fitri RA. 2020. Short Communication: Detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products using Real-Time PCR Multiplex Method. Asian J Nad Prod Biochem 21: 17-20. The detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products was carried out using Multiplex Real-Time PCR (qPCR) in the Microbiology and Molecular Biology Laboratory at the Indonesian Food and Drug Authority in Gorontalo. The purpose of this study was to provide alternative testing methods for food products circulating in the market. The sample consisted of 25 samples of processed meat products spike with Salmonella typhimurium ATCC 14028 phase 2 and Listeria monocytogenes ATCC 7644 phase 2. The method used in the study was qPCR analysis using the SYBR Green method, while DNA isolation used the direct PCR method. Data analysis was carried out based on Cycle threshold and Melting temperature based on two main criteria. Cycle threshold (Ct) analysis determines the Ct value of the sample and comparing it with the control. Melting temperature (Tm) analysis determines the temperature at which 50% of double-stranded DNA changed to a single standard and comparing it with the melting temperature of positive control. The results showed Salmonella typhimurium ATCC 14028 in the processed meat was detected at an average Ct value of 10.34, and a Tm value of 85.70. The presence of Listeria monocytogenes ATCC 7644 in the samples was recognized at an average Ct value of 14.04, and an average Tm value of 80.07. It can be concluded that the real-time multiplex PCR method can be used to detect Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 by using the melting curve (Tm) analysis.

Hematological, serum biochemical and histopathological effects of selected herbs and combinations on Trypanosoma brucei infected West African dwarf sheep

Olatunde OA, Jegede HO, Ameen SA. 2021. Hematological, serum biochemical and histopathological effects of selected herbs and combinations on Trypanosoma brucei infected West African dwarf sheep. Asian J Nat Prod Biochem 19: 10-16. The present work was carried out to study the hematological, serum biochemical and histopathological effects of selected herbal diets on trypanosome infected West African dwarf sheep. Five herbal incorporated diets were tested where A is control diet, B is Citrus aurantifolia; C is Ocimum gratissimum, D is Vernonia amygdalina + Citrus aurantifolia and E is Ocimum gratissimum + Vernonia amygdalina. Sheep on Diets E, A, C, B, and D consumed 856.4g/d, 830g/d, 750.0g/d. 734.0g/d and 687.0g/d respectively showing more acceptability to diet E. Additionally, sheep on Diet E was highest in RBC, WBC, Hb, lymphocyte, PCV and basophils. The least hematological indices were recorded for the animals on Control diet (A) showing that all herbal diets improved blood indices of the infected sheep. The ALP concentration was significantly different (17.000i.u/l) in Diet B compared to other Diets while the least ALP was noted for sheep on Diets A and B. Herbs and combinations used in this study significantly improved both hematological and pathological lesions seen in experimental trypanosomosis caused by Trypanosoma brucei infection in sheep. Best results were gotten from Diet E combinations.

Review: Microbial induced mineralization of calcium carbonate for self-healing concrete

Elkhateeb WA, Elnahas MO, Daba GM. 2021. Review: Microbial induced mineralization of calcium carbonate for self-Healing concrete. Asian J Nat Prod Biochem 19: 1-9. Low-cost solutions achieving concrete self-healing are attracting researchers attention. Generally, concrete self-healing mechanism has been so far accomplished by three approaches: autogenous healing, encapsulation of polymeric material, and microbial induced mineralization of calcium carbonate. Microbial approach seems like an attractive potent, relatively cheap way to achieve concrete self-healing. Hence, this review elucidates the microbial concrete self-healing mechanisms, and compare the roles of fungal and bacterial mediated self-healing concrete.

Anti-inflammatory activities of ethanol leaves extract and solvent fractions of Zehneria scabra (Cucurbitaceae) in rodents

Belay R, Makonnen E. 2018. Anti-inflammatory activities of ethanol leaves extract and solvent fractions of Zehneria scabra (Cucurbitaceae) in rodents. Biofarmasi J Nat Prod Biochem 18: 42-56. Zehneria scarba (L.f.) Sond is one of the medicinal plants used in folkloric medicine of Ethiopia for years to treat various inflammatory disorders. The present study was aimed to validate the anti-inflammatory activity of crude 70% ethanol leaves extract (70EE) against a sub-acute model and further evaluate the solvent fractions (AF, BF, and CF) in an acute (carrageenan-induced paw edema), sub-acute (formaldehyde induced arthritis) and chronic (cotton pellet induced granuloma) inflammatory models. The 70EE was first prepared by maceration, and the fractions were obtained by sequential partitioning with chloroform and n-butanol from the aqueous suspension of crude 70EE. The test groups, then, received 100, 200 and 400 mg/kg of the crude 70EE or the fractions (AF, BF, and CF) at the same dose levels, whereas positive controls received aspirin (200mg/kg) or dexamethasone (0.5mg/kg) and negative controls received vehicle (2% tween 80 or distilled water, 10 mL/kg). All tested doses of the crude 70EE showed significant inhibition of formaldehyde induced arthritis at the 10th day of treatment, on which the 400mg/kg dose showed the maximum anti-arthritic effect (%A = 60.5; p < 0.001). In the carrageenan-induced paw edema, all the three fractions showed a statistically significant effect, in fact, with different onset and magnitude. In this model, the AF was found to be the most active fraction, and the 400mg/kg dose demonstrated the maximum effect (%A = 76.25; p < 0.001) at 5h post-induction, which is much better than the effect of aspirin at the dose employed. The overall order of efficacy in inhibiting the exudative component of carrageenan-induced paw edema was found to be AF> BF> CF. The AF was also found to be the most active fraction in inhibiting the exudative component of chronic inflammation in the cotton pellet induced granuloma model, where the maximum effect (%A = 43.10, p < 0.001) was exhibited by a dose of 400mg/kg. The AF was also the most active fraction in inhibiting formaldehyde induced arthritis, in which the BF and CF relatively showed a comparable effect throughout day 4-10. On the contrary, in the cotton pellet induced granuloma model, the CF was found to be the most active fraction in inhibiting the proliferative and granulomatous component of chronic inflammation, and the overall order of effectiveness was found to be CF> AF> BF. Besides, 400mg/kg of CF demonstrated the maximum inhibition of granuloma formation (%A = 55.52; P < 0.001). The phytochemical analysis revealed the differential distribution of secondary metabolites into the three fractions, which either singly or in concert appeared to be responsible for the observed effects. The data obtained from the present study collectively indicate that the extract and fractions of leaves of Z.scabra possessed a significant anti-inflammatory activity, upholding the folkloric use of the plant.

Phytochemical composition and antimicrobial activity of Ochna thomasiana

Muema MJ, Machocho AK, Gikonyo NK. 2018. Phytochemical composition and antimicrobial activity of Ochna thomasiana. Biofarmasi J Nat Prod Biochem 18: 29-41. Infectious diseases are the leading cause of mortality worldwide despite the vigorous campaigns that have been made to combat them. This study aimed at the determination and evaluation of the biological activities of Ochna thomasiana Engl. & Gilg ex Gilg. Here, we screened the plant extracts and tested for their antibacterial activity against Salmonella typhi (clinical isolate), Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis, and Staphylococcus aureus. Various chromatographic techniques separated and isolated the active compounds from this plant. The extracts were purified using silica gel, column chromatography (CC), Sephadex gel, and preparative thin-layer chromatography (PTLC). Structure characterization was determined using standard spectroscopic methods: Infrared (IR), ultraviolet (UV) spectroscopy, mass spectroscopy (MS) and proton nuclear magnetic resonance (1H NMR), distortionless enhancement by polarization transfer (DEPT), carbon-13 nuclear magnetic resonance (13C NMR), coherence spectroscopy (COSY), heteronuclear multiple bond coherence (HMBC) and heteronuclear single quantum coherence (HSQC). Compounds identified from the extracts are Lophirone A (18), afzelone D dimethyl ether (20), calodenone (17), a mixture of stigmasterol (74) and ?-sitosterol (23) and 3?-acetyl-24-ethylfriedelane (75). The stem and root bark of methanol crude extracts showed high activity against the Gram-positive bacteria. Lophirone A, afzelone D dimethyl ether, and 3?-acetyl-24- ethylfriedelane showed high activity against S. aureus. The results showed the root of O. thomasiana contains biflavonoids, and some sterols as its constituents and their antimicrobial activity are significant and is a lead towards the development of antimicrobial agents. The essential bioactive compounds and the antimicrobial activity of the crude extracts of this plant confirms its use in traditional medicine. However, there is a need to test the effectiveness of the crude extracts and isolated compounds via in vivo and in vitro evaluation. The plant species should be cultivated using good agricultural practices for medicinal plants for the future assessment of their activity against pathogens.

Consumption patterns, perceptions and total carotenoids, iron and zinc contents of yellow flesh cassava

Duah EA, Steiner-Aseidu M, Danquah AO. 2018. Consumption patterns, perceptions, and total carotenoids, iron, and zinc contents of yellow flesh cassava. Biofarmasi J Nat Prod Biochem 18: 13-28. The main objective was to investigate consumption patterns of white cassava, knowledge, perceptions and the nutritive profiles of yellow flesh cassava roots and leaves and its relationship to the recommended daily allowance for vitamin A. Total carotenoids, iron and zinc contents in fresh yellow cassava leaves, roots and products (gari, konkonte, boiled cassava roots and leaves), antioxidant activity and the in vitro bioaccessibility of the carotenoids were determined using standard methods. A cross-sectional survey was carried out to identify the consumption patterns of white cassava, the knowledge, and the perception of yellow flesh cassava among Ghanaians. Total carotenoids (T.C), iron (Fe) and zinc (Zn) for fresh roots ranged from 4.73±0.11 to 10.11±0.18 µg/g; 87.35±3.18 to 146.25±1.20 mg/100g; 0.30± 0.01 to1.55±0.07 mg/100g, respectively. T.C, Fe and Zn for fresh leaves ranged from 792.93±0.98 to 2649.20±29.10 µg/g; 118.35±0.070 to 07182.05± mg/100g; 3.75±0.64 to 15.50±0.14 mg/100g respectively. T.C for gari was between 3.21±2.79 and 7.39±1.06 µg/g, iron 118.75±0.64 to 181.85±2.05 mg/100g and zinc, 0.25±0.07 to 0.80±0.14 mg/100g. Iron and zinc contents for the kokonte samples ranged from 101.45±0.64 to 116.30± 0.14 mg/100g and 0.15±0.07 to 0.70±0.01 mg/100g, however, T.C was not detectable. T.C for boiled roots were between 1.22±0.05 and 2.14±0.11µg/g. Iron content for the samples ranged from 118.75±0.64 to 181.85±2.05 mg/100g and for zinc, 0.60±0.01 to 1.30±0.01 mg/100g. T.C for boiled leaves was between 524.39±9.89 and 1323.5±15.6 µg/g, iron; 95.90±0.01 and 148.75±2.76 mg/100g and zinc 0.60±0.01-1.30±0.01mg/100g. In vitro bio-accessibility of carotenoids for boiled roots had the highest of 104.42±0.88%, gari had the highest value of 57.22±9.01%, and boiled leaves 0.28±0.01%. Gari recorded the highest frequency of consumption. 36.6% of the respondents knew yellow flesh cassava. 51.2% were willing to accept yellow flesh cassava. Fermentation, drying, roasting, and boiling retained some carotenoids after processing, but solar drying over a long period completely degraded carotenoids in yellow cassava roots. Cassava leaves had higher retention of carotenoids, but cassava roots had more bio-accessible carotenoids. Carotenoids in yellow flesh cassava leaves had antioxidant properties that have the potential to help combat free radicals in the body. The knowledge and “willingness to accept” yellow cassava was low among Ghanaians. Yellow cassava will serve as a promising source of provitamin A.

Screening and partial characterization of d-endotoxins from some local Bacillus thuringiensis isolates for insecticidal activity against the spotted stem borer

Kimani PG, Nyambaka H, Kasina M. 2018. Screening and partial characterization of d-endotoxins from some local Bacillus thuringiensis isolates for insecticidal activity against the spotted stem borer. Biofarmasi J Nat Prod Biochem 17: 21-38. Prolonged use of synthetic chemical pesticides is environmentally undesirable, causing the rapid development of resistance among insect pests. Kenya has unexplored potential in controlling lepidopteran pests by using proteinous delta-endotoxins sourced from local isolates of a naturally occurring bacterium, Bacillus thuringiensis Berliner (Bt). This study attempted to identify the insecticidal proteins in some Kenyan Bt isolates, characteristic of Cry1 and/or Cry2 proteins. It also aimed to test their efficacy as affected by different temperatures and their specificity on an invasive and prevalent lepidopteran stem borer, Chilo partellus (Swinhoe). Delta-endotoxin crystals were isolated and purified from cultures of twenty unidentified local Bt isolates using froth floatation and centrifugation. Total protein in the resulting suspensions was quantified using the Bradford assay method, and the approximate protein yield was 3.14 ±0.084 mg/mL of nutrient broth culture with a purity level of 54.8 % ±15.3 %. Leaf-dip bioassays used for testing the efficacy of the d-endotoxins against C. partellus. Among the isolates evaluated, Bt 44 and Bt 48 had the most potent d-endotoxin crystals towards the 1st instar larvae, leading to mortality of 62.6 % and 64.8 % respectively after 72 h. The effect of the d-endotoxins' concentration and temperature on larval mortality was examined for 72 hours at temperatures of 24°C, 27°C and 31°C and levels of 0.01 mg/mL, 0.1 mg/mL and 1.0 mg/mL. The resulting LC50 was 52.3 µg/mL and 42.0 µg/mL, while LT50 values were 76.7h and 60.9h for Bt 44 and Bt 48, respectively. Higher efficacy found at 24°C and 31°C than at 27°C, an indication that these d-endotoxins are tolerable for local conditions where temperatures are higher than in temperate regions. The relationship between concentration and temperature was significant for d-endotoxins of Bt 48 but not those of Bt 44. A major protein component of the d-endotoxins had a molecular weight Mr ~ 130 kDa, which generates a trypsin-resistant core of Mr ~ 70 kDa. Cry protein analysis detected more Cry1 in Bt 44 than Bt 48 ?-endotoxins and no Cry2 in either. However, cry gene analysis using PCR detected the presence of both cry1 and cry2 genes in the DNA of Bt 44 but none in Bt 51, a negative control from toxicity tests against the pest. The chromatographic analysis revealed some differences in the elution profiles of d-endotoxins of both Bt 44 and Bt 48, an indication that there may be different types and amounts of the Cry toxins in the crystals or even novel proteins. These findings indicate that the two local Bt isolates expressed Cry1, and probably Cry2 proteins can control C. partellus and may, therefore, become promising sources for ?-endotoxins for biopesticide development for controlling the pest.