Nat Prod Research Articles

Review: The endless nutritional and pharmaceutical benefits of the Himalayan gold, Cordyceps; Current knowledge and prospective potentials

Abstract. Elkhateeb WA, Daba GM. 2020. Review: The endless nutritional and pharmaceutical benefits of the Himalayan gold, Cordyceps; Current knowledge and prospective potentials. Biofarmasi J Nat Prod Biochem 18: 70-77. As a traditional medicine, Cordyceps has long been used in Asian nations for maintaining vivacity and boosting immunity. Numerous publications on various bioactivities of Cordyceps have been investigated in both in-vitro as well as in vivo studies. Nevertheless, the role of Cordyceps is still arguable whether it acts as food supplement for health benefits or a real healing drug that can be prescribed in medicine. The Cordyceps industry has developed greatly and offers thousands of products, commonly available in a global marketplace. In this review, focus will be on introducing the ecology of Cordyceps and their classification. Moreover, elucidation of the richness of extracts originated from this mushroom in nutritional components was presented, with description of the chemical compounds of Cordyceps and its well-known compounds such as cordycepin, and cordycepic acid. Furthermore, highlights on natural growth and artificial cultivation of famous Cordyceps species were presented. The health benefits and reported bioactivities of Cordyceps species as promising antimicrobial, anticancer, hypocholesterolemic, antioxidant, antiviral, anti-inflammatory, organ protective agent, and enhancer for organ function were presented.

Evaluation of the antiproliferative activity of red propolis hydroalcoholic extract and its fractions obtained by partition

Abstract. Squarisi IS, De Freitas KS, Lemes DC, Ccana-Ccapatinta GV, Mejia JAA, Bastos JK, Veneziani RCS, Ambrosio SR, Tavares DC. 2018. Evaluation of the antiproliferative activity of red propolis hydroalcoholic extract and its fractions obtained by partition. Biofarmasi J Nat Prod Biochem 18: 66-69. The present study aimed to evaluate the cytotoxicity of red propolis hydroalcoholic extract (RPHE) and its fractions obtained by partition, hexanes (HF), dichloromethane (DF), ethyl acetate (AF) and n-butanol (BF), on tumor and non-tumor cell lines. For this purpose, the XTT colorimetric assay was performed on human lung fibroblasts (GM07492A, non-tumor cell), breast adenocarcinoma (MCF-7), glioblastoma (U343) and cervix adenocarcinoma (HeLa) cells. The results showed that RPHE, HF and DF presented not only cytotoxic potential to all tumor cell lines but also to normal cell line, indicating selectivity absence. HF presented the lowest IC50 (half minimal inhibitory concentration; 33.8-133.3 µg/mL), with significant difference from those observed for RPHE (137.0-262.7 µg/mL). BF and AF revealed an IC50 which was higher than 1250 µg/mL in all cell lines. The results showed that red propolis has substances with antiproliferative activity, indicating that its hexanes fraction may have substances with antitumor potential.

Detection of Salmonella typhimurium ATCC 14028 in supplement health product liquid preparation using Real-Time PCR (qPCR)

Abstract. Sophian A, Purwaningsih R, Lukita BL, Ningsih EC. 2018. Detection of Salmonella typhimurium ATCC 14028 in supplement health product liquid preparation using Real-Time PCR (qPCR). Biofarmasi J Nat Prod Biochem 18: 61-65. Detection of Salmonella typhimurium ATCC 14028 using Real-Time PCR (qPCR) on health supplement products was carried out in the microbiology and molecular biology testing laboratory of the Food and Drug Supervisory Office in Gorontalo. The purpose of this study was to provide an alternative testing method reference in the testing of liquid supplement health supplement products in the market. The sample consisted of 35 samples of liquid supplement health supplements spike with positive control of Salmonella typhimurium ATCC 14028 phase 2. The method used in the study was qPCR analysis using the SYBR Green method, whereas DNA isolation using the direct PCR method. Data analysis was performed based on 2 main criteria: (i) Ct (Cycle threshold) analysis, which is looking at the value of the sample Ct and comparing it with controls and (ii) analysis of melting temperature (Tm), which is the melting point at the temperature at which melt occurs and comparing the melting point to the positive control. The results showed that in the sample, Salmonella typhimurium ATCC 14028 was detected at an average Ct value of 14.43, and an average Tm value of 86.05, for the specificity, LOD and positive control tests were all amplified. For negative controls, Ct and Tm values ​​were not detected. Based on these data it can be concluded that real-time PCR (qPCR) can be used to detect Salmonella typhimurium ATCC 14028 in liquid supplement health supplement products.

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

Abstract. 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

Abstract. 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

Abstract. 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.

Survey of the bacterial diversity at two coastal beaches in Guyana

Abstract. Prashad L, Daniel R, Ram M. 2020. Survey of the bacterial diversity at two coastal beaches in Guyana. Biofarmasi J Nat Prod Biochem 18: 1-8. The #63 Beach, Berbice and Marriott Beach, Kingston Seawall, Guyana were surveyed for halophilic bacteria present in its waters. NaCl tolerance, temperature tolerance and antimicrobial activity of isolates against Staphylococcus aureus and Bacillus sp. were tested. Samples of 100 mL were taken from the Shore, 5.0 m, and 10.0 m depth from both locations. The samples were plated and examined for the growth of bacteria of different pigmentation. A total of 4 halophilic isolates were found; 3 from Marriott Beach (Isolates A, B, and G) and 1 (Isolate M) from the #63 Beach. Isolates were of yellow, light orange and pink pigmentation, the 3 isolates from Marriott Beach were, Gram-negative and cocci while the one isolates from #63 Beach were Gram-positive and cocci. The optimum salinity tolerance for the Isolate G from Marriott Beach was 1.5M NaCl, Isolate M from #63 Beach 1.0M NaCl, Isolate B from Marriott Beach 1.0M NaCl and Isolate A from, Marriott Beach 0.5M NaCl. The optimum temperature for the growth of the isolates was 37°C. The isolates had no antimicrobial activity against S. aureus and Bacillus sp.

Protective effects of Curcuma longa rhizomes ethyl acetate extract against alcohol induced oxidative stress and nephrotoxicity in female Wistar rats

Abstract. Eteng OE, Moses CA, Enobong J, Akamo AJ, Akinloye DI, Ugbaja RN, Akinloye OA. 2020. Protective effects of Curcuma longa rhizomes ethyl acetate extract against alcohol induced oxidative stress and nephrotoxicity in female Wistar rats. Biofarmasi J Nat Prod Biochem 21: 5-12. This study aimed to evaluate the protective effect of Curcuma longa Linn. (syn. Curcuma domestica Val.) rhizomes ethyl acetate extract (CLREAE) facing alcohol-induced oxidative stress and nephrotoxicity. Thirty female (30) Wistar rats were categorized randomly into six groups. Groups 1, 2, 3, 4, 5 and 6 were treated with normal saline; 20% ethanol; 100 mg of CLREAE + 20% ethanol; 200 mg of CLREAE + 20%; 350 mg of CLREAE + 20% ethanol and 350 mg of CLREAE respectively for 14 days. A significant (p<0.05) decrease in the SOD, CAT and GPx activities and GSH concentration of rat treated with only 20% ethanol were found when compared to the normal control group, whereas a significant (P<0.05) increase in the groups pretreated with different doses of the CLREAE were also found when compared to groups with only 20% ethanol treatment. Thus, comparing to the normal control group, treatment with the CLREAE fetched a significant (p<0.05) decrease in the renal biomarkers (creatinine and urea). Whilst, comparing to the groups with 20% methanol treatment, a significant (p<0.05) increase happened in the groups pretreated with different doses of the CLREAE. There was a significant (p<0.05) decrease on Kidney MDA level in rats pretreated with different doses of CLREAE compared with the normal control. It was shown in the results of the histology that there was a physiologic recovery in the kidney tissues as groups were treated with different doses of the CLREAE. Evidenced by reduced necrosis of tubular and glomerular epithelial, the signs of protection against toxicity were found on the rats. The study suggested that through in vivo free radical scavenging ability, the CLREAE has protective effects against alcohol-induced oxidative stress and nephrotoxicity in female Wistar rats.

Cytotoxic and genotoxic effects of the ent-kaurenoic acid and ent-kaurenoic acid enriched Mikania glomerata extract in V79

Abstract. Ferreira NH, Ribeiro AB, Morais MD, Peixoto AM, Bernardino MA, Moreira MR, Soares ACF, Heleno VCG, Veneziani RCS, Tavares DC. 2018. Cytotoxic and genotoxic effects of the ent-kaurenoic acid and ent-kaurenoic acid-enriched Mikania glomerata extract in V79. Biofarmasi J Nat Prod Biochem 17: xxxx. The ent-kaurenoic acid-rich extract from Mikania glomerata Sprengel was effective to inhibit the formation of Streptococcus mutans biofilm. In view of the biological potential of this extract and its major component, the present study was carried out to evaluate the safety of the ent-kaurenoic acid-rich extract from Mikania glomerata Sprengel and ent-kaurenoic acid alone in an in vitro test system. The results showed that the ent-kaurenoic acid-rich extract from Mikania glomerata Sprengel was cytotoxic at concentrations up to 40.0 μg/mL. Genotoxic effects were observed in cell cultures treated with the highest concentrations tested of ent-kaurenoic acid-rich extract from Mikania glomerata Sprengel (10.0 and 15.0 µg/mL) and ent-kaurenoic acid alone (2.5, 5.0 and 7.5 µg/mL) when compared to the control group. Therefore, the ent-kaurenoic acid-rich extract from Mikania glomerata Sprengel demonstrated cytotoxicity and genotoxicity effects at the highest concentrations tested, while ent-kaurenoic acid showed to be genotoxic at the same concentrations present in ent-kaurenoic acid-rich extract from Mikania glomerata Sprengel in V79 cells. These results demonstrate that the ent-kaurenoic acid should be responsible, at least in part, of the genotoxicity of ent-kaurenoic acid-rich extract from Mikania glomerata Sprengel.

Antioxidant and alpha-glucosidase inhibitory activities of Euchresta horsfieldii

Abstract. Prihantini AI, Krisnawati, Setyayudi A. 2019. Antioxidant and alpha-glucosidase inhibitory activities of Euchresta horsfieldii. Biofarmasi J Nat Prod Biochem 17: 61-64. Euchresta horsfieldii, known as pranajiwa, is a medicinal plant that is widely grown in Bali and West Nusa Tenggara, Indonesia. Its seeds or fruits are commonly used for body freshness and stamina. The present study aimed to investigate the biological activities of leaves, root, stem, fruits, seeds of the E. horsfieldii. Antioxidant, alpha-glucosidase inhibitory activities and total phenolic compound were evaluated from methanolic extracts of all parts of E. horsfieldii. The result showed that leaf extract of E. horsfieldii exhibited the highest antioxidant activity with IC50 215.11±08.06 µg/mL. Meanwhile, the root extract had the highest alpha-glucosidase inhibitory activity and total phenolic compound with IC50 29.76±13.17 µg/mL and 763±0.01 mg GAE/100mg dry extract, respectively. In conclusion, the study suggested that E. horsfieldii is potential as natural source of antioxidant and alpha-glucosidase inhibitor agents.

Nutritional value and safety of castor bean (Ricinus communis) seeds detoxified in solid-state fermentation by Pleurotus ostreatus

Abstract. Adebayo OC, Ogidi CO, Akinyele BJ. 2019. Nutritional value and safety of castor bean (Ricinus communis) seed detoxified in solid-state fermentation by Pleurotus ostreatus. Biofarmasi J Nat Prod Biochem 17: 51-60. The nutrient and antinutrient contents of unfermented castor seeds (UCS), castor seeds fermented with Pleurotus ostreatus (CPF) and “Ogiri,” a naturally fermented condiment from castor seeds (CSF) were determined. Proximate analyses of all the samples were carried out using methods of Association of Official Agricultural Chemists. The biosafety of the samples was carried out using the animal model. The raw castor seeds (UCS) had the highest carbohydrate (61.04%), ash (6.02%), fats (6.65%), fiber (6.62%), and calcium (0.30 mg/100g). Castor seeds fermented with Pleurotus ostreatus (CPF) had the highest protein content of 20.47%, magnesium of 7.16 mg/100g, alkaloids (7.40 mg/g) and saponins (6.69 mg/g) . CSF had the highest zinc (0.69 mg/100g). CPF had the lowest tannin (0.05 mg/g). All the essential amino acids increased significantly in the fermented samples. Tryptophan was absent in UCS but present in CSF and CPF with values of 0.78 mg/100g and 1.15 mg/100g, respectively. The hematological analysis of the rats fed CPF had the highest WBC of 5.43×109, which indicated a positive immunomodulatory effect. Hence, this study revealed that Pleurotus ostreatus degraded the toxic compounds in castor seeds to a large extent and enhanced the nutritional contents of the final product.

The potential of white-oyster mushroom (Pleurotus ostreatus) as antimicrobial and natural antioxidant

Abstract. Egra S, Kusuma IW, Arung ET, Kuspradini H. 2018. The potential of white-oyster mushroom (Pleurotus ostreatus) as antimicrobial and natural antioxidant. Biofarmasi J Nat Prod Biochem 16: 17-23. White-oyster mushroom (Pleurotus ostreatus) is a favorite meal in Indonesia. Previously this fungus was known as a useless plant, but after the nutrition is known, everything changes. People tried to cultivate it because the nutrients contents are very good for body health. Therefore, to support this added value in the field of health, especially antimicrobials and antioxidants, this research needs to be done. This research used successive extraction with hexane solvent, acetate ethyl, ethanol, water, and crude ethanol by antimicrobial assay, antioxidants assay (DPPH), total antioxidant content, total phenolic content. The highest results on barrier antimicrobial test which occurred against Candida albicans bacteria was 47.60 % with 100 ppm concentration. While, on antimicrobial assay using Propionibacterium acnes bacteria, there was no significant inhibition. Regarding the antioxidant test against DPPH, the result showed the occurrence of free radical by 25 % on water extraction at the concentration of 100 ppm. Continuously, the total antioxidant content assay showed the ethyl acetate had the highest value of 368.708 mg gae/g. The results of the total content phenolic assay showed the solvent hexane had the value of 78.495 mg gae/g. These findings indicated that mushroom has an active phenolic compound with no contribution to impede its working on Candida albicans assay.

Investigation of the hemostatic effects of freeze-dried extracts of selected Kenyan plants

Abstract. Makunyi EG, Bukachi F, Waweru P. 2019. Investigation of the hemostatic effects of freeze-dried extracts of selected Kenyan plants. Biofarmasi J Nat Prod Biochem 17: 39-46. This study aims to investigate the effect and mechanism of action of freeze-dried extracts of Tridax procumbens, Terminalia brownii, Euphorbia tirucalli, and Asphillia africana on hemostasis. Freeze-dried extract of the selected plants was prepared and dose determined for the study. Twelve male New Zealand white rabbits were randomly allocated to two groups (control and test). Blood was collected under standard procedures. Duke's method was used for the bleeding time while the capillary method was used for clotting time. ACL Elitepro machine was used to do prothrombin time and activated partial thromboplastin time. Thromboelastography was done for the most potent extracts. Data were analyzed using independent t-test and results were presented as mean± standard error of means. Differences were considered to be significant if P < 0.05. The results showed that the percentage yield of the extract was; Tridax procumbens (0.8%), Terminalia brownii (0.5%), Euphorbia tirucalli (0.2%) and Aspillia pluriseta (1.3%). Bleeding and clotting time: The bleeding time was reduced by freeze-dried leaf extract of Tridax procumbens (p = 0.0068) and by freeze-dried bark extract of Terminalia brownii (p=0.0068). Freeze-dried leaf extract of Asphillia africana increased the bleeding time (p=0.01). The clotting time was reduced by freeze-dried leaf extract of Tridax procumbens (p = 0.038), the freeze-dried bark extract of Terminalia brownii (p=0.043) and by freeze-dried stem extract of Euphorbia tirucalli (p=0.01). Prothrombin and activated partial thromboplastin time: The prothrombin time was reduced by freeze-dried leaf extract of Tridax procumbens (p = 0.004), the freeze-dried bark extract of Terminalia brownii (p<0.001) and Freeze-dried stem extract of Euphorbia tirucalli (p=0.001). Activated partial thromboplastin time was reduced by freeze-dried leaf extract of Tridax procumbens (p< 0.001), the freeze-dried bark extract of Terminalia brownii (p<0.001) and by freeze-dried stem extract of Euphorbia tirucalli (p<0.001). The results for thromboelastogrpahy showed that four parameters of thromboelastography were tested. Freeze-dried leaf extract of Tridax procumbens reduced the r time (p =0.04), k time (p=0.04) and maximum amplitude (p =0.026) but increased the alpha angle (p =0.01). The freeze-dried bark extract of Terminalia brownii did not have statistically significant differences in thromboelastography variables.

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

Abstract. 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.

Nutritional composition of aquatic plants and their potential for use as animal feed: A case study of the Lower Volta Basin, Ghana

Etse WJ, Annang T, Ayivor JS. 2018. Nutritional composition of aquatic plants and their potential for use as animal feed: a case study of the Lower Volta Basin, Ghana. Biofarmasi J Nat Prod Biochem 9: 99-112. The study was conducted to determine the nutritional composition of selected dominant aquatic plants and their significant effect on the chemical and physical characteristics of the water. Aquatic plants namely Nymphaea lotus, Typha australis, Ipomoea aquatica, and Scirpus cubensis were collected, identified and authenticated at the Ghana Herbarium. The proximate nutritional compositions of these plants were measured using the standard procedure outlined in the Association of Official Analytical Chemist (AOAC 2002). Water and sediment quality analyses of some physicochemical variables were also carried out using processes described in the standard methods for water and wastewater examination. The results showed that nutrient composition such as the crude protein, ether extracts, ash content, and nitrogen-free extracts was significantly higher than the corresponding constituents in Panicum maximum used as a control for the study. The findings also indicated that levels of heavy metals in all plants fell within the WHO/FAO standards for metals in vegetables and food. The effects of the physicochemical parameter of water also revealed that pH, nitrate, turbidity, DO, and BOD levels were found significantly different from the control site. The level of heavy metal in the sediment samples revealed significant variations in the distribution of the metals, with Zn showing the most significant difference and Pb the least with a mean level of 7.5±0.86 mg/L and 0.4±0.03 mg/L respectively. These plant species suggests having a high nutritive potential and indicates their possible use as mixed ingredients in animal feed. Exploitation of these aquatic plants for animal feed would be a step towards better utilization of these plants help in the management of aquatic plants within the basin.

Characterization of flavonoids from candidate striga grass in controlling diet legumes Cicer arietinum and Vigna radiata

Akumu AA, Nyambaka H, Kuate SP, Torto B.2018. Characterization of flavonoids from candidate striga grass in controlling diet legumes Cicer arietinum and Vigna radiata.Biofarmasi J Nat Prod Biochem 16: 83-98. In Africa, intercropping of some legumes with cereal plants has been noticed to generate exceptional reduction of Striga invasion and enhanced production of cereal crop. Desmodium uncinatum administers Striga invasion in intercrop with cereals through an allelopathic technique, which includes postsprouting accretion inhibitors emanated from the roots. Having a potential to be a new technique for grass administering, Allelopathy is a natural and environmentally friendly method. A standardized profiling technique based on Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) and Liquid Chromatography coupled with Mass Spectrometry (LC/MS) was utilized to determine flavonoids in extracts of chickpea (Cicer arietinum) and mung bean (Vigna radiata), potential trap plants for Striga grass. Nine flavonoids involving mono-and diglycosyl derivatives of fisetin, baicalein, rhamnetin, isorhamnetin, formononetin, quercetin, isosakuranetin, and sakuranetin were temporarily distinguished. The obtained phenolics were available at concentrations higher than 0.001% of the dry materials. Many of these phenolic mixtures have been claimed to possess human health advantages. These glycosylated flavones are noted for the first time in these two species. The comprehensive analysis of the polar secondary metabolites in these leguminous plants was useful for comprehending their inhibitory chemistry and proposed biosynthesis by C-glucoside characteristics. All the segregated and described mixtures in the diet legume C. arietinum were O-glucosylated. While, Food legume V. radiata should be observed further to see if it can attain Desmodium’s allelopathic ability since it possesses inhibitory chemistry and proposed biosynthesis by C-glucosylation.

Evaluation of antimicrobial and antioxidant attributes of Tanzanian honey from two agro-ecological areas

Luvanda FT, Lyimo ME. 2018. Evaluation of antimicrobial and antioxidant attributes of Tanzanian honey from two agroecological areas. Biofarmasi J Nat Prod Biochem 16: 69-82. In order to appraise the characteristic of Tanzanian honey based on antioxidant attributes and antimicrobial activity on bacteria and fungi in relation to its physicochemical attributes, the study was conducted. The examined honey was sampled from two kinds of bees namely stinging and stingless honey bees and was gathered from the western area (Tabora and Shinyanga) and the central area (Singida and Dodoma). They were examined for their antimicrobial activity, anti-oxidant attributes (total phenol, vitamin C) and physicochemical attributes of pH, color, pH, sugar, and minerals. 54 % of tested honey samples inhibited microbial growth but the remaining 46% did not inhibit the growth. Microbial impediment areas range from 8.5 to 14.16 mm (stinging honey bee) and 10.56-15.13 mm (stingless honey bee). Honey yielded by stinging honey bee in the cities of Shinyanga, Bukombe, and Nzega and the cities of Singida and Issuna is more effective in impeding microbial growth than that of other cities. Candida albicans is more sensitive (23.1%) to honey of stingless honey bee, followed by Staphylococcus saprophyticus (33%), Salmonella typhi (32%), Escherichia coli (19.2%) and Aspergillus flavus (16.0%). The substance of antioxidant, total phenol and vitamin C in honey of stingless honey bee were importantly (P <0.05) higher than that of honey of stinging honey bee. Antioxidants (FRAP) ranged from 322.16-973.57 µMFe (II) /100g, total phenols was from 13.87-33.55 mg /100g and vitamin C was from 2.54-10.99 mg /100. At pH value, there was no substantial dissimilarity (P> 0.05) between the two samples of honey. Potassium possesses the highest value while Zn possesses the lowest value in samples of honey of stinging and stingless honey bee. The gradation color of honey of stingless honey bee ranged from extra light amber-light amber while that of honey of stinging bee ranged from white water-light amber. There was a positive and substantial correlation (P <0.001) between the color of honey, antioxidants, total phenol, and mineral substance. Stingless honey bee's honey from the western region is superior to stinging honey bee's honey in antioxidant and antimicrobial attributes and therefore is prompted to be utilized as a valuable diet product, preventive and curative medicine.

The hydroxyproline content of fish bone gelatin from Indonesian Pangasius catfish by enzymatic hydrolysis for producing the bioactive peptide

Abstract. Atma Y, Lioe HN, Prangdimurti E, Seftiono H, Tahufik M, Fitriani D, Mustopa AZ. 2018. The hydroxyproline content on fish bone gelatin from Indonesian Pangasius catfish by enzymatic hydrolysis for producing of bioactive peptide. Biofarmasi J Nat Prod Biochem 16: 64-68. Gelatins has been widely used in food, medicines, cosmetics, photography industries. In the food industry, gelatin used as food additives and functional foods. The applications of gelatin as a functional food due to their bioactivity in a form of peptides. Bioactive peptides from gelatin are mostly obtained through enzymatic hydrolysis processes. This study was conducted to measure the hydroxyproline content of gelatin bone of Indonesian Pangasius catfish (Pangasius sutchi) before and after enzymatic hydrolysis. The hydroxyproline is one of the dominant amino acids in gelatin. Gelatin hydrolysis was carried out using a flavourzyme at concentration of 6% and the incubation series of 0, 4, 6 and 8 hours. The standard concentration of hydroxyproline was used in range of 0-1 μg. The results showed that the linear curve of the hydroxyproline standard solution was y=0.0554x+0.0406, with the coefficient of determination (R²) = 0.9435. The incubation time of enzymatic hydrolysis (6% enzyme concentration) affected the hydroxyproline content. The hydroxyproline from fish bone gelatin was 18.91±2.87 mg/mL, 63.81±1.28 mg/mL, 46.21±1.28 mg/mL and 37.64 ± 0.64 mg/mL respectively during 0, 4, 6 dan 8 h incubation time. This hydroxyproline content was significantly different at 95% confidence level from each treatment time.

The effect of jengkol (Archidendron pauciflorum) fruit peel ethanolic extract to heart histologic of rat induced by streptozotocin

Abadi SA, Illiyyin Z, Rachmadina JR, Malini DM. 2018. The effect of jengkol (Archidendron pauciflorum) fruit peel ethanolic extract to heart histologic of rat induced by streptozotocin. Biofarmasi J Nat Prod Biochem 16: 59-63. Indonesia is ranked 7th out of 10 countries with the highest diabetic patients in the world. Diabetes mellitus is a disease that can cause heart disorders and adults who suffer from DM are four times more likely to develop heart disease. Jengkol fruit peel has been used traditionally as a drug for diabetes mellitus. The aim of this research was to know the effect of fruit peel ethanolic extract of jengkol (Archidendron pauciflorum) (JFPEE) on rat (Rattus norvegicus) heart histological structure and to obtain effective dose from JFPEE. This research used experimental method in laboratory with Completely Random Design (CRD) using 6 treatments and 4 replications. Treatment was given for 14 consecutive days consisting of negative control, positive control, comparison (glibenclamide dose 10 mg/kg BW), P1, P2, and P3 (JFPEE dose 385, 770, and 1.540 mg/kg BW). Diabetic induction was performed with streptozotocin dose 65 mg/kg BW in female wistar rat except for negative control group. The parameters that observed were number of necrosis and cell damage score, including fat degeneration, hydropic degeneration, and inflammatory cell. The obtained data were analyzed by ANOVA-Tukey's test with 95% confidence level using SPSS version 21 for Windows. The result of histological structure showed that number of necrosis and cell damage score in group of rats treated with JFPEE dose 385 mg/kg BW (174.25±6.34; 1.25±0.50) were not significantly different from the negative control rats (172.00±7.62; 1.00±0.00). The effective dose of JFPEE that can repair the damage of heart cell’s rat induced by streptozotocin was 385 mg/kg BW.

Novel flax orbitide derived from genetic deletion

BackgroundFlaxseed orbitides are homodetic plant cyclic peptides arising from ribosomal synthesis and post-translation modification (N to C cyclization), and lacking cysteine double bonds (Nat Prod Rep 30:108-160, 2013). Screening for orbitide composition was conducted on the flax core collection (FCC) grown at both Saskatoon, Saskatchewan and Morden, Manitoba over three growing seasons (2009-2011). Two flax (Linum usitatissimum L.) accessions ‘Hollandia’ (CN 98056) and ‘Z 11637’ (CN 98150) produce neither [1−9-NαC]-linusorb B2 (3) nor [1−9-NαC]-linusorb B3 (1). Mass spectrometry was used to identify novel compounds and elucidate their structure. NMR spectroscopy was used to corroborate structural information.ResultsExperimental findings indicated that these accessions produce a novel orbitide, identified in three oxidation states having quasimolecular ion peaks at m/z 1072.6 (18), 1088.6 (19), and 1104.6 (20) [M + H]+ corresponding to molecular formulae C57H86N9O9S, C57H86N9O10S, and C57H86N9O11S, respectively. The structure of 19 was confirmed unequivocally as [1−9-NαC]-OLIPPFFLI. PCR amplification and sequencing of the gene coding for 18, using primers developed for 3 and 1, identified the putative linear precursor protein of 18 as being comprised of the first three amino acid residues of 3 (MLI), four conserved amino acid residues of 3 and/or 1 (PPFF), and the last two residues of 1 (LI).ConclusionComparison of gene sequencing data revealed that a 117 base pair deletion had occurred that resulted in truncation of both 3 and 1 to produce a sequence encoding for the novel orbitide precursor of 18. This observation suggests that repeat units of flax orbitide genes are conserved and suggests a novel mechanism for evolution of orbitide gene diversity. Orbitides 19 and 20 contain MetO and MetO2, respectively, and are not directly encoded, but are products of post-translation modification of Met present in 18 ([1−9-NαC]-MLIPPFFLI).