In-vitro colonic fermentation of Kakadu plum (Terminalia ferdinandiana) fruit powder: Microbial biotransformation of phenolic compounds and cytotoxicity.

Kakadu plum (Terminalia ferdinandiana) (KP) is an indigenous fruit used as a functional ingredient in powdered form. Three KP doses (1, 2.5 and 5 g) were digested in a dynamic in vitro gut digestion model over 48 h. Faecal water digests from the colonic reactors were assessed for total soluble polyphenols (TSP), ferric reducing antioxidant power (FRAP), phenolic metabolites and short-chain fatty acids (SCFAs). Effects of digests on cell viability were tested against Caco-2 intestinal and HepG2 hepatic cells. All doses of KP fermentation produced castalagin, corilagin, chebulagic acid, chebulinic acid, and gallic acid. TSP and FRAP significantly increased in 5 g KP digests at 0 and 48 h of fermentation. SCFA concentrations significantly increased after 48 h. Cytotoxic effects of 2.5 and 5 g KP digests diminished significantly after 12 h. Overall, colonic fermentation increased antioxidant activity and polyphenolic metabolites of 5 g KP powder for 48 h.

Bioactive Properties of Kakadu Plum-Blended Products.

Kakadu plum (Terminalia ferdinandiana), endemic to Australia, is growing in popularity due to its high levels of vitamin C and strong antioxidant properties. In this study, Kakadu plum fruit powder was used as a functional food ingredient with other plant materials to develop value-added products to enhance their nutritional and commercial value. The present study determined the bioactive properties of nine products, including three Kakadu plum fruit powder samples produced from different processing batches and five Kakadu plum-blended products. Vitamin C, the total phenolic content, and the ellagic acid content were determined. Bioactive properties such as antioxidant, antidiabetic, and antimicrobial assays were also performed. Cytotoxicity was tested to obtain more specific product information regarding food safety. Kakadu plum-blended products showed lower cytotoxicity and lower bioactive properties (antioxidant and antibacterial activities) in comparison to Kakadu plum powder. However, overall, most of the bioactive properties were shown to be higher in the blends when compared with the commercial blueberry powder as a benchmark antioxidant product. Therefore, there is great potential for Kakadu plum to contribute to the growing functional food and ingredient markets.

Investigating the Efficacy of Tasmannia lanceolata Extract in Inactivating Fungi and Prolonging the Shelf Life of Date Fruit.

Date palm (Phoenix dactylifera L.) is one of the world's oldest cultivated plants. Post-harvest losses of date palm due to fungal contamination reached up to 50% of the total production. This study aimed to investigate the potential of the extract of Tasmanian pepper leaf (TPL) and the non-thermal treatment of photosensitization mediated by curcumin in reducing the fungal contamination and enhancing the shelf life of date palm. In the in vivo storage study, the dates were treated with three different concentrations of TPL extract 12.5, 25, and 50 µg/mL and stored at 30 °C. The findings obtained for the treatment with TPL extract exhibited potent antifungal activity against most of the tested fungi, where minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) were < 25 µg/mL for polygodial, the bioactive compound in TPL. The shelf life of date palm treated by 50 µg/mL polygodial was extended up to 21 days, thrice as much as the untreated controls. In contrast, a lower concentration of TPL extract (25 µg/mL polygodial) revealed up to a 15-day shelf life extension compared to untreated dates (7 days). The results obtained from the study suggested that using TPL extracts against pathogenic and spoilage fungi occurring in fresh date fruits is a promising treatment for the shelf life extension of fresh date fruits at room temperature.

Effects of drying methods and maltodextrin on vitamin C and quality of Terminalia ferdinandiana fruit powder, an emerging Australian functional food ingredient.

BACKGROUND: Terminalia ferdinandiana, common name Kakadu plum (KP), fruit is a valuable source of vitamin C, and its concentration can be used as a quality index of KP products, such as dried fruit powder. The present study investigated the effects of two drying methods (freeze-drying and oven-drying) and the addition of maltodextrin (0-25%) on vitamin C, Maillard products, and overall quality of KP fruit powder. RESULTS: Freeze-drying was a better dehydration technique than oven-drying in retaining vitamin C, reducing the formation of non-enzymatic browning and oxidation products, and improving powder colour (P < 0.05). Non-enzymatic browning products (furfural and 5-hydroxymethyl furfural) were generated in the oven-dried samples as a function of heating and high water activity. Maltodextrin acted as a vitamin C stabilizer in protecting vitamin C from oxidation, and significantly improved the colour attributes of the final dry products. Incorporation of 10-15% maltodextrin could reduce the percentage loss of vitamin C from 8.1% to 3.4% and 18.9% to 11.4% (compared with the control) during freeze-drying and oven-drying, respectively. Scanning electron micrographs revealed differences in the microstructures of the KP powder processed by the two drying methods with different levels of maltodextrin. Multivariate data analysis (principal component analysis) showed separation between the oven-dried and freeze-dried samples, and also suggested that addition of maltodextrin of 7.5-10% and 10-15% are effective for preserving vitamin C and other quality properties of the freeze- and oven-dried KP powder samples, respectively. CONCLUSION: The results obtained are important for the KP industry, including Indigenous enterprises, in selecting the most appropriate drying method for KP fruit in terms of quality and sustainability. © 2021 Society of Chemical Industry.

High throughput screening to determine the antibacterial activity of Terminalia ferdinandiana (Kakadu plum): A proof of concept.

Different types of susceptibility tests are available to identify antimicrobial activity, including the disc agar diffusion and broth micro-dilution methods. In recent years, high throughput screening (HTS) methods have been considered and evaluated as an efficient method to rapidly monitor the antimicrobial potential of a wide range of plant products. The objective of this study was to test the ability of a 96-well plate reader as HTS method to evaluate the antimicrobial potential of extracts of Terminalia ferdinandiana (Kakadu plum). The main changes observed in the UV-VIS spectra of the bacteria samples were related to the biochemical and chemical compounds that might originate from the effect of the T. ferdinandiana extracts and the bacteria. Partial least squares discriminant analysis (PLS-DA) allowed the correct classification of samples according to the concentration of extract added to the culture (e.g. high, medium and low). The results of this study indicated that might be possible to record changes in the UV-VIS spectra associated with the interactions between bacteria and T. ferdinandiana extracts using a 96-well plate reader. The method was able to detect or differentiate between live and dead bacteria based on the UV-VIS spectra as a function of the addition of the T. ferdinandiana extracts.

In vitro Bioaccessibility and Intestinal Absorption of Selected Bioactive Compounds in Terminalia ferdinandiana.

Terminalia ferdinandiana (or Kakadu plum), a native Australian fruit with potential health benefits, contains bioactive compounds such as ellagic acid (EA), ascorbic acid (AA) and calcium, and antinutrients such as oxalic acid (OA). However, few is known about the biological fate of these compounds following ingestion; therefore, the aim of this study was to evaluate in vitro bioaccessibility and intestinal absorption of T. ferdinandiana compounds using the INFOGEST static digestion model and Caco-2-HT29-MTX-E12 intestinal absorption model. No significant changes (p > 0.05) were observed in total AA content throughout in vitro digestion, whereas bioaccessibility of EA, OA, and calcium increased significantly from 33, 72, and 67% in the gastric phase to 48, 98, and 90% in the intestinal phase, respectively. The intestinal absorption study revealed variable rates of movement across the cell barrier. Findings reveal novel and important insights for the prediction of in vivo bioavailability of selected T. ferdinandiana compounds.

Impact of polyphenol-rich extracts of Terminalia ferdinandiana fruits and seeds on viability of human intestinal and liver cells in vitro.

Terminalia ferdinandiana (Kakadu plum) is a native Australian fruit consumed by Indigenous Australians for centuries. Commercial interest in T. ferdinandiana has increased in recent years due to its high vitamin C content, however, food safety assessments are lacking. To explore the safety of extracts prepared from T. ferdinandiana using different solvents, in vitro cell viability of undifferentiated and differentiated Caco-2, HT29-MTX-E12, and HepG2 cells was measured using the CyQUANT® NF Cell Proliferation Assay. Changes to cell viability produced IC50 values between 3650 and 14400 µg/mL for all extracts and cell lines tested with HepG2 cells impacted the most by T. ferdinandiana extracts, followed by HT29-MTX-E12 cells, and undifferentiated and differentiated Caco-2 cells. Different solvents also produced extracts with variable effects on cell viability that were dependent on tissue source, however, extracts from seedcoats appeared to impact cell viability less than fruit extracts. The IC50 values for ellagic acid, an abundant phytochemical in T. ferdinandiana, varied from 1190 to 2390 µg/mL across different cells and were significantly lower than extract IC50 values. Findings from this study will help to inform future safety studies, select which solvents to use when preparing T. ferdinandiana extracts, and decide whether fruit flesh should be separated from seeds during extract preparation.

Interactions Between Phytochemicals and Minerals in Terminalia ferdinandiana and Implications for Mineral Bioavailability.

Oxalic and phytic acid are phytochemicals considered to be anti-nutritional factors as they are predominantly found as oxalates and phytates bound to minerals like calcium and potassium. Studies have associated excessive oxalate consumption with increased urinary excretion of oxalate (hyperoxaluria) and calcium oxalate kidney stone formation, and excessive phytate consumption with decreased bioaccessibility and bioavailability of certain minerals and reduced utilization of dietary protein. However, other studies suggest that dietary consumption of phytate may be beneficial and inhibit formation of calcium oxalate kidney stones. In light of these conflicting reports, dietary intake of oxalate and phytate enriched plants should be considered in relation to potential health outcomes following consumption. Terminalia ferdinandiana is one such plant and is investigated here with respect to oxalate, phytate, and mineral contents. Assessment of oxalate and phytate contents in T. ferdinandiana fruit, leaf, and seedcoat tissues through hydrolysis into acid forms revealed oxalic acid contents ranging from 327 to 1,420 mg/100 g on a dry weight (DW) basis whilst phytic acid contents ranged from 8.44 to 121.72 mg/100 g DW. Calcium content in the different tissues ranged from 131 to 1,343 mg/100 g. There was no correlation between oxalic acid and calcium, however a significant, positive correlation was observed between phytic acid and calcium (r = 0.9917; p < 0.001), indicating that tissues rich in phytic acid also contain higher levels of calcium. The high content of phytic acid in comparison to oxalic acid in T. ferdinandiana fruit found in this study and the dietary significance of this in terms of calcium bioavailability, needs to be investigated further.

Antioxidant Rich Extracts of Terminalia ferdinandiana Inhibit the Growth of Foodborne Bacteria.

Terminalia ferdinandiana (Kakadu plum) is a native Australian plant containing phytochemicals with antioxidant capacity. In the search for alternatives to synthetic preservatives, antioxidants from plants and herbs are increasingly being investigated for the preservation of food. In this study, extracts were prepared from Terminalia ferdinandiana fruit, leaves, seedcoats, and bark using different solvents. Hydrolysable and condensed tannin contents in the extracts were determined, as well as antioxidant capacity, by measuring the total phenolic content (TPC) and free radical scavenging activity using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay. Total phenolic content was higher in the fruits and barks with methanol extracts, containing the highest TPC, hydrolysable tannins, and DPPH-free radical scavenging capacity (12.2 ± 2.8 g/100 g dry weight (DW), 55 ± 2 mg/100 g DW, and 93% respectively). Saponins and condensed tannins were highest in bark extracts (7.0 ± 0.2 and 6.5 ± 0.7 g/100 g DW). The antimicrobial activity of extracts from fruit and leaves showed larger zones of inhibition, compared to seedcoats and barks, against the foodborne bacteria Listeria monocytogenes, Bacillus cereus, Methicillin resistant Staphylococcus aureus, and clinical isolates of Pseudomonas aeruginosa. The minimum inhibitory concentration and minimum bactericidal concentration in response to the different extracts ranged from 1.0 to 3.0 mg/mL. Scanning electron microscopy images of the treated bacteria showed morphological changes, leading to cell death. These results suggest that antioxidant rich extracts of Terminalia ferdinandiana fruits and leaves have potential applications as natural antimicrobials in food preservation.

Antioxidant-Rich Extracts of Terminalia ferdinandiana Interfere with Estimation of Cell Viability.

The impact of plant extracts and phytochemicals on in vitro cell viability is usually assessed by employing cell viability assays dependent upon the activity of dehydrogenase enzymes. The CellTiter 96® AQueous One Solution Cell Proliferation Assay (CellTiter) was used to measure cell viability in response to antioxidant-rich extracts of Terminalia ferdinandiana fruits. Conflicting results were obtained from this assay whereby higher concentrations of extracts significantly increased cell viability compared to lower concentrations. Intrinsic reductive potential was observed in a cell-free system when extracts were added directly to the CellTiter assay reagent. To confirm this effect in a similar cell proliferation assay, we employed the CellTiter-Blue® Cell Viability Assay and again observed increased viability with increased concentrations of the extracts and direct reduction of the assay reagent by the extracts in cell-free systems. In the search for a cell proliferation assay that would not be directly affected by the plant extracts, we identified the CyQUANT® NF Cell Proliferation Assay that is based on the estimation of DNA content in viable cells. Cell viability decreased with increasing concentrations of the extracts. Accordingly, the results of the present study indicated that cell viability assays reliant upon dehydrogenase activity may lead to false positive results when testing antioxidant-rich plant extracts with intrinsic reductive potential, and alternative cell viability assays should be used to measure the cell viability.

Chemical and Nutritional Composition of Terminalia ferdinandiana (Kakadu Plum) Kernels: A Novel Nutrition Source.

Terminalia ferdinandiana (Kakadu plum) is a native Australian fruit. Industrial processing of T. ferdinandiana fruits into puree generates seeds as a by-product, which are generally discarded. The aim of our present study was to process the seed to separate the kernel and determine its nutritional composition. The proximate, mineral and fatty acid compositions were analysed in this study. Kernels are composed of 35% fat, while proteins account for 32% dry weight (DW). The energy content and fiber were 2065 KJ/100 g and 21.2% DW, respectively. Furthermore, the study showed that kernels were a very rich source of minerals and trace elements, such as potassium (6693 mg/kg), calcium (5385 mg/kg), iron (61 mg/kg) and zinc (60 mg/kg) DW, and had low levels of heavy metals. The fatty acid composition of the kernels consisted of omega-6 fatty acid, linoleic acid (50.2%), monounsaturated oleic acid (29.3%) and two saturated fatty acids namely palmitic acid (12.0%) and stearic acid (7.2%). The results indicate that T. ferdinandiana kernels have the potential to be utilized as a novel protein source for dietary purposes and non-conventional supply of linoleic, palmitic and oleic acids.

Antinociceptive and neuropharmacological activities of methanol extract of Phoenix sylvestris fruit pulp.

Fruits of Phoenix sylvestris Roxb. (Arecaceae) are used to treat back pain, toothache, headache, arthritis, nervous debility and as sedative. The aim of this study was to evaluate the antinociceptive and neuropharmacological activities of methanol extract of P. sylvestris fruit pulp (MEPS). The antinociceptive activity of MEPS was evaluated by heat-induced (hot plate, tail immersion test) and chemical-induced pain models (acetic acid-induced writhing, formalin-induced nociception, glutamate-induced nociception and paw edema test). The effect of MEPS on central nervous system (CNS) was studied using hole cross test, open field test, sodium thiopental-induced sleeping time and elevated plus maze test. MEPS showed strong, significant and dose-dependent antinociceptive activity in all heat-induced and chemical-induced pain models at all experimental doses. Involvement of opioid receptor mediated analgesia was evident from the reversal of analgesic effect by naloxone. MEPS also showed reduced locomotor activity in both hole cross and open field tests. The increase in sleeping time in sodium thiopental-induced sleeping test and anxiolytic activity in elevated plus maze test were also significant. So, it is evident that MEPS possesses strong central and peripheral antinociceptive activity as well as CNS depressant, sedative and anxiolytic activity. The results justify the ethnomedicinal use of P. sylvestris fruit in different painful conditions and CNS disorders.

Awareness regarding the importance of calcium and vitamin D among the undergraduate pharmacy students in Bangladesh.

BACKGROUND: Calcium and vitamin D are two important micronutrients required for maintaining proper bone health. Previous works intended to determine the status of these micronutrients in local population have reported that the people in Bangladesh are at high risk of calcium insufficiency and hypovitaminosis D related health complications. Lack of awareness and insufficient knowledge of the essentiality of these two nutrients are assumed to cause this problem in Bangladesh. The present study was designed and conducted to establish a basic understanding on the level of gap of knowledge and awareness among pharmacy students at undergraduate level in Bangladesh. FINDINGS: A total of 713 students of Bachelor of Pharmacy course participated in the study. The students were asked about basic idea related to calcium and vitamin D and the disorders due to their deficiency, name of common foods containing calcium and vitamin D, their perception regarding the essentiality of the said nutrients etc. It was found that most of the students were familiar with the importance of calcium (98.9%) and vitamin D (99.3%) in bone health. 82.2% students know about the term osteoporosis. Unfortunately, 10.7% and 18.8% students failed to mention at least one food that is rich in calcium and vitamin D, respectively. Most of the students got familiar about the nutrients from their teachers (48.9%) and textbooks (32.8%). CONCLUSION: Being a student of pharmacy, the students should have more comprehensive knowledge about calcium and vitamin D. The present study indicates that the pharmacy students have lack of knowledge about calcium and vitamin D and thus it can be clearly predicted that the condition of general people may be worse.

Antinociceptive, antidiarrheal, and neuropharmacological activities of Barringtonia acutangula.

CONTEXT: Barringtonia acutangula (L.) Gaertn. (Lecythidaceae) has been used in folk medicine in the treatment of arthralgia, chest pain, dysmenorrhea, inflammation, hemorrhoids, diarrhea, and also in psychological disorders. OBJECTIVE: To investigate the antinociceptive, antidiarrheal, and neuropharmacological effect of the methanol extract of B. acutangula leaves and seeds in mice. MATERIALS AND METHODS: The extracts (200 and 400 mg/kg; p.o.) were tested for antinociceptive activity by acetic acid-induced writhing, hot plate and tail immersion models; castor oil- and magnesium sulphate-induced diarrheal models were used to evaluate antidiarrheal activity whereas hole cross and open field models were employed for testing neuropharmacological activity. RESULTS: Both extracts exhibited significant antinociceptive effect (p < 0.001) in acetic acid and heat induced pain models in a dose-dependent manner. The extracts prolonged the latency period to the thermal stimuli in both hot plate and tail immersion test. The extracts also showed significant inhibition of defecation (p < 0.001, 0.01) in both diarrheal models. Again, the spontaneous motor activity was decreased (p < 0.001) by the extracts in both hole cross and open field test. DISCUSSION AND CONCLUSION: The results of this study suggest that the methanol extracts of B. acutangula leaves and seeds possess good antinociceptive, antidiarrheal, and central nervous system (CNS) depressant activities. This study validates the use of this plant in traditional medicine.

Antinociceptive activity of methanol extract of flowers of Impatiens balsamina.

ETHNOPHARMACOLOGICAL RELEVANCE: Impatiens balsamina Linn. (Balsaminaceae), an annual herb locally called "Dopati", is cultivated as an ornamental garden plant in Bangladesh. Flowers of the plant are used in folk medicine to treat lumbago, neuralgia, burns and scalds. AIM OF THE STUDY: This study evaluated the antinociceptive effect of the methanol extract of I. balsamina flowers (MIB). MATERIALS AND METHODS: The extract was evaluated for antinociceptive activity using chemical- and heat-induced pain models such as acetic acid-induced writhing, hot plate, tail immersion and formalin test. To verify the possible involvement of opioid receptor in the central antinociceptive effect of MIB, naloxone was used to antagonize the effect. The effect of MIB on central nervous system (CNS) was also studied using hole cross and open field tests. RESULTS: MIB demonstrated strong and dose-dependent antinociceptive activity in all the chemical- and heat-induced mice models (p<0.05). These findings imply the involvement of both peripheral and central antinociceptive mechanisms. The use of naloxone confirmed the association of opioid receptors in the central antinociceptive effect. MIB also showed significant central nervous system depressant effect (p<0.05). CONCLUSION: This study reported the peripheral and central antinociceptive activity of the flowers of I. balsamina and rationalized the traditional use of the flower in the treatment of different painful conditions.