Water temperature is a major abiotic driver, controlling the rates and nature of biochemical reactions and subsequently affecting the physiology of marine organisms. However, relatively little is known about the implications of heat stress or predicted ocean climate change on marine secondary metabolites. The predatory gastropod Dicathais orbita is a useful model organism for climate change and natural product studies. Here we determine the upper thermal limit (CTMax) of D. orbita and investigate the effects of thermal stress on the bioactive compounds stored in their biosynthetic organ, the hypobranchial gland. Two CTMax experiments were undertaken, along with a static heat stress experiment where whelks were exposed to an elevated temperature of 30°C for one week, compared to a 20°C seawater control. An additional 35-day ocean climate change experiment used combinations of temperature (ambient: 23°C and future: 25°C) and pCO2 (ambient: ~380 ppm and future: ~765 ppm). The impacts on secondary metabolites in all experiments were assessed using liquid chromatography-mass spectrometry. The mean CTMax of the whelks, from the northern limit of their distribution, was found to be 35.2°C using a rapid temperature increase rate of 1°C/1 h, but was only 30.6°C when a gradual heating rate of 1°C/12 h was used. The overall composition of the secondary metabolites was significantly affected by heat stress in all four experiments, but not by elevated pCO2 in the ocean climate change experiment. The proportion of the choline ester murexine was significantly reduced in heat-stressed snails compared to the controls. Tyrindoxyl sulphate was significantly reduced under prolonged exposure to future temperature, whereas the relative abundance of the oxidation product, 6-bromoisatin significantly increased with elevated temperature exposure. Despite the fact that intertidal gastropods like D. orbita might be able to buffer the impact of external temperatures within the predicted future range, this study provides evidence that ocean warming could have significant implications for secondary metabolite production and/or storage in marine invertebrates. Impacts on bioactive molecules with multifunctional ecological roles could have implications for predator populations with possible flow on effects in some marine communities.

Waste from food production can be re-purposed as raw material for usable products to decrease industrial waste. Coffee pulp is 29% of the dry weight of coffee cherries and contains caffeine, chlorogenic acid, trigonelline, diterpenes and fibre. We investigated the attenuation of signs of metabolic syndrome induced by high-carbohydrate, high-fat diet in rats by dietary supplementation with 5% freeze-dried coffee pulp for the final 8 weeks of a 16-week protocol. Coffee pulp decreased body weight, feed efficiency and abdominal fat; normalised systolic blood pressure, left ventricular diastolic stiffness, and plasma concentrations of triglycerides and non-esterified fatty acids; and improved glucose tolerance in rats fed high-carbohydrate, high-fat diet. Further, the gut microbiota was modulated with high-carbohydrate, high-fat diet and coffee pulp supplementation and 14 physiological parameters were correlated with the changes in bacterial community structures. This study suggested that coffee pulp, as a waste from the coffee industry, is useful as a functional food for improving obesity-associated metabolic, cardiovascular and liver structure and function, and gut microbiota.

The pulp of the purple mangosteen, Garcinia mangostana, is a popular tropical fruit but the rind containing xanthones such as α-mangostin together with procyanidins and anthocyanidins is usually discarded as waste. However, this rind has been used in South-East Asia for diarrhoea, dysentery, skin infections and wounds. As xanthones have reported anti-inflammatory and antioxidant responses, this study has determined the bioactive compounds and evaluated the effects of G. mangostana rind on physiological, metabolic, liver and cardiovascular parameters in rats with diet-induced metabolic syndrome. Rats fed a diet with increased simple sugars and saturated fats developed obesity, hypertension, increased left ventricular stiffness, dyslipidaemia and fatty liver. Administration of G. mangostana rind as 5% of the food to rats with diet-induced metabolic syndrome gave a dose of 168 mg/kg/day α-mangostin, 355 mg/kg/day procyanidins, 3.9 mg/kg/day anthocyanins and 11.8 mg/kg/day hydroxycitric acid for 8 weeks which reduced body weight and attenuated physiological and metabolic changes in rats including decreased abdominal fat deposition, decreased abdominal circumference and whole-body fat mass, improved liver structure and function and improved cardiovascular parameters such as systolic blood pressure, left ventricular stiffness and endothelial function. These responses were associated with decreased infiltration of inflammatory cells, decreased deposition of collagen in both heart and liver and decreased mean adipocyte size in retroperitoneal adipose tissues. We conclude that, in rats with diet-induced metabolic syndrome, chronic intake of G. mangostana rind decreased infiltration of inflammatory cells which decreased physiological, metabolic, liver and cardiovascular symptoms.

Saskatoon berry (Amelanchier alnifolia) is a potential functional food containing anthocyanins and flavonols, as well as ellagitannins and phenolic acids. We have determined the potential therapeutic effects of Saskatoon berry in diet-induced metabolic syndrome. Nine- to ten-week-old male Wistar rats were randomly assigned to four groups. Two groups were fed on control diets, either corn starch (C) or high-carbohydrate, high-fat diet (H) respectively, for 16 weeks. Two further groups were fed on C or H diet for 16 weeks with Saskatoon berry powder added to the diet for the final 8 weeks (CSSK, HSSK). After 16 weeks, H rats showed symptoms of metabolic syndrome, including increased body weight, visceral adiposity, systolic blood pressure, cardiac fibrosis, plasma concentrations of triglycerides and non-esterified fatty acids, and plasma activities of alanine transaminase and aspartate transaminase. Saskatoon berry intervention normalised body weight and adiposity, improved glucose tolerance, decreased systolic blood pressure, improved heart and liver structure and function with decreased infiltration of inflammatory cells, and decreased plasma total cholesterol. Further, Saskatoon berry normalised liver expression of hexokinase 1 and glycogen phosphorylase and increased glucose 6-phosphatase relative to H rats. These results suggest that Saskatoon berry regulates glycolysis, gluconeogenesis and glycogenesis to improve metabolic syndrome.

Coffee brewing produces spent coffee grounds as waste; few studies have investigated the health benefits of these grounds. This study investigated responses to spent coffee grounds in a diet-induced rat model of metabolic syndrome. Male Wistar rats aged 8-9 weeks were fed either corn starch-rich diet or high-carbohydrate, high-fat diet for 16 weeks, which were supplemented with 5% spent coffee grounds during the last 8 weeks. Rats fed non-supplemented diets were used as controls. High-carbohydrate, high-fat diet-fed rats developed metabolic syndrome including abdominal obesity, impaired glucose tolerance, dyslipidemia, and cardiovascular and liver damage. Body weight, abdominal fat, total body fat mass, systolic blood pressure, and concentrations of plasma triglycerides and non-esterified fatty acids were reduced by spent coffee grounds along with improved glucose tolerance and structure and function of heart and liver. Spent coffee grounds increased the diversity of the gut microbiota and decreased the ratio of Firmicutes to Bacteroidetes. Changes in gut microbiota correlated with the reduction in obesity and improvement in glucose tolerance and systolic blood pressure. These findings indicate that intervention with spent coffee grounds may be useful for managing obesity and metabolic syndrome by altering the gut microbiota, thus increasing the value of this food waste.

Carrageenans are thickening and gelling agents that may provide health benefits. Iota (ι)-carrageenan, a linear sulfated polysaccharide, is produced by the red seaweed, Sarconema filiforme. This study investigated the potential of this seaweed as a functional food for the reversal of metabolic syndrome and possible mechanisms. Male Wistar rats were divided into four groups in a 16-week protocol: corn starch diet-fed rats (C); C rats supplemented with 5% S. filiforme for the last 8 weeks (CSF); high-carbohydrate, high-fat diet-fed rats (H); and H rats supplemented with 5% S. filiforme for the last 8 weeks (HSF). S. filiforme was produced in tank-based aquaculture yielding 27 g dry weight/day/m2 of culture area. H rats developed obesity, hypertension, dyslipidaemia, glucose intolerance, fatty liver and increased left ventricular collagen deposition. S. filiforme supplementation decreased body weight, abdominal and liver fat, systolic blood pressure, plasma total cholesterol concentrations, and plasma activities of alanine transaminase and aspartate transaminase. S. filiforme supplementation modulated gut microbiota without changing the Firmicutes to Bacteroidetes ratio. S. filiforme improved symptoms of high-carbohydrate, high-fat diet-induced metabolic syndrome in rats. Possible mechanisms include a reduced infiltration of inflammatory cells into organs as well as prebiotic actions in the gastrointestinal tract.

Metabolic syndrome, especially obesity, increases the risk of type 2 diabetes and cardiovascular disease. We hypothesised that green coffee will attenuate metabolic, cardiovascular and liver parameters in high-carbohydrate high-fat diet-fed rats. Male Wistar rats (8–9 weeks old) were divided into 6 groups and fed for 16 weeks with either corn starch diet (C), C with either 5% green or decaffeinated green coffee in food for the last 8 weeks, high-carbohydrate high-fat diet (H) or H with either 5% green or decaffeinated green coffee in food for the last 8 weeks. Green coffee contained chlorogenic acids, trigonelline, caffeine and diterpenoids; decaffeinated green coffee contained these compounds but no caffeine. Green coffee attenuated body weight, systolic blood pressure, inflammation in the heart and liver and diastolic stiffness without improving glucose sensitivity or plasma lipids. We suggest that chlorogenic acids, trigonelline and diterpenoids in green coffee attenuate diet-induced abnormalities in heart and liver.

The secondary metabolite Tyrian purple, also known as shellfish purple and royal purple, is a dye with historical importance for humans. The biosynthetic origin of Tyrian purple in Muricidae molluscs is not currently known. A possible role for symbiotic bacteria in the production of tyrindoxyl sulphate, the precursor to Tyrian purple stored in the Australian species, Dicathais orbita, has been proposed. This study aimed to culture bacterial symbionts from the purple producing hypobranchial gland, and screen the isolates for bromoperoxidase genes using molecular methods. The ability of bromoperoxidase positive isolates to produce the brominated indole precursor to Tyrian purple was then established by extraction of the culture, and analysis by liquid chromatography–mass spectrometry (LC–MS). In total, 32 bacterial isolates were cultured from D. orbita hypobranchial glands, using marine agar, marine agar with hypobranchial gland aqueous extracts, blood agar, thiosulphate citrate bile salts sucrose agar, and cetrimide agar at pH 7.2. These included 26 Vibrio spp., two Bacillus spp., one Phaeobacter sp., one Shewanella sp., one Halobacillus sp. and one Pseudoalteromonas sp. The two Bacillus species were the only isolates found to have coding sequences for bromoperoxidase enzymes. LC–MS analysis of the supernatant and cell pellets from the bromoperoxidase producing Bacillus spp. cultured in tryptone broth, supplemented with KBr, confirmed their ability to produce the brominated precursor to Tyrian purple, tyrindoxyl sulphate. This study supports a potential role for symbiotic Bacillus spp. in the biosynthesis of Tyrian purple.

The broad utilisation of imidacloprid (IMI) in agriculture poses an increasing risk to aquatic organisms. However, the potential impacts on commercially important shellfish and chemical residues after exposure, are yet to be assessed. We investigated the levels of IMI in Sydney rock oyster (SRO) tissue during a three-day uptake and four-day depuration cycle using liquid chromatography-mass spectrometry. IMI was absorbed from the water, with significantly higher concentrations in the adductor muscles than the gills and digestive glands. Depuration was also fast with a significant drop in tissue concentrations after one day in clean water and complete elimination from all tissues except the digestive gland after four days. The distribution of IMI in SRO after direct exposure using mass spectrometry imaging demonstrated uptake and spatially resolved metabolism to hydroxyl-IMI in the digestive gland and IMI-olefin in the gills. We assessed the effects of IMI on filtration rate (FR), acetylcholinesterase (AChE) activity in the gills, and gene expression profiles in the digestive gland using transcriptomics. Exposure to 2 mg/L IMI reduced the FR of oysters on the first day, while exposure to 0.5 and 1 mg/L reduced FR on day four. IMI reduced the gill AChE activity and altered the digestive gland gene expression profile. This study indicates that commercially farmed SRO can uptake IMI from the water, but negative impacts were only detected at concentrations higher than currently detected in estuarine environments and the chemical residues can be effectively eliminated using simple depuration in clean water.

Metabolic syndrome is a cluster of disorders that increase the risk of cardiovascular disease and diabetes. This study has investigated the responses to rind of yellow mangosteen (Garcinia dulcis), usually discarded as waste, in a rat model of human metabolic syndrome. The rind contains higher concentrations of phytochemicals (such as garcinol, morelloflavone and citric acid) than the pulp. Male Wistar rats aged 8–9 weeks were fed either corn starch diet or high-carbohydrate, high-fat diet for 16 weeks, which were supplemented with 5% freeze-dried G. dulcis fruit rind powder during the last 8 weeks. We characterised metabolic, cardiovascular, liver and gut microbiota parameters. High-carbohydrate, high-fat diet-fed rats developed abdominal obesity, hypertension, increased left ventricular diastolic stiffness, decreased glucose tolerance, fatty liver and reduced Bacteroidia with increased Clostridia in the colonic microbiota. G. dulcis fruit rind powder attenuated these changes, improved cardiovascular and liver structure and function, and attenuated changes in colonic microbiota. G. dulcis fruit rind powder may be effective in metabolic syndrome by appetite suppression, inhibition of inflammatory processes and increased fat metabolism, possibly related to changes in the colonic microbiota. Hence, we propose the use of G. dulcis fruit rind as a functional food to ameliorate symptoms of metabolic syndrome.

Davidson’s plum is a native Australian fruit used traditionally as food and medicine. The fruit contains anthocyanins as glucosides and sambubiosides of cyanidin and peonidin (691 mg/100 g of dried pulp) with rutin and quercetin glycosides (193 mg/100 g). Dietary supplementation of Davidson’s plum at approximately 8 mg anthocyanins/kg/day in rats attenuated the signs of metabolic syndrome induced by high-carbohydrate, high-fat diet. Davidson’s plum reduced visceral fat accumulation, total abdominal fat weight, size of retroperitoneal adipocytes, and plasma triglycerides and non-esterified fatty acids, normalised blood pressure, reduced left ventricular stiffness, decreased infiltration of inflammatory cells in both left ventricle and liver, decreased collagen deposition in heart, and reduced both fat vacuoles in liver and obesity-induced degeneration of knee cartilage. There were no changes in glucose tolerance with treatment. Davidson’s plum reduced colonic Clostridiaceae spp, and increased Turicibacter spp. and Akkermansia muciniphila. Our findings indicate that Davidson’s plum is a potential complementary treatment for metabolic syndrome.

The Australian marine mollusc Dicathais orbita has been identified as a functional food with potential for use in the prevention of colon cancer. This study investigated the effect of cooking on the biochemical profile of the edible flesh and extracts containing anticancer brominated indoles. The whelk flesh was high in protein (> 75 mg/100 mg dry weight) and there was a significant increase in protein and amino acids after cooking, associated with a loss of moisture and lipids. The flesh also has a high proportion of polyunsaturated fatty acids (> 49%), with omega-3: omega-6 around 1. The flesh contains unusually high levels of Docosapentaenoic acid and over 1000 mg/100 g serve of Eicosapentaenoic acid and Docosahexaenoic acid. Cooking resulted in a quantitative decrease in the bioactive compounds, however the main anticancer compound 6-bromoisatin can be retained after boiling. This provides evidence that cooking does not negatively impact the functional food properties of these muricids whelks.

Garcinia humilis is a fruit known as achachairú. It is native to South American countries such as Bolivia, Peru, and Brazil, but it is also cultivated as achacha in northern Australia. The aim of this study was to determine the phytochemicals in achacha rind and pulp and to investigate these components as potential treatments for the symptoms of metabolic syndrome. Both rind and pulp contain procyanidins and citric acid rather than hydroxycitric acid. Male Wistar rats (8–9 weeks old) were fed with either high-carbohydrate, high-fat, or corn starch diets for 16 weeks. Intervention groups were fed with either diet supplemented with 1.5% G. humilis rind powder or 2.0% G. humilis pulp for the last 8 weeks of the protocol. Rats fed a high-carbohydrate, high-fat diet exhibited hypertension, dyslipidemia, central obesity, impaired glucose tolerance, and non-alcoholic fatty liver disease. G. humilis rind decreased systolic blood pressure, diastolic stiffness, left ventricular inflammatory cell infiltration, and collagen deposition in high-carbohydrate, high-fat diet-fed rats. However, there was no change in glucose tolerance, body weight, or body composition. Therefore, G. humilis rind, usually a food by-product, but not the edible pulp, showed potential cardioprotection with minimal metabolic changes in a rat model of diet-induced metabolic syndrome.

Muricidae molluscs are the source of a valuable purple dye that was traded as a luxury item in the Mediterranean region and by the late Byzantine was reserved for royalty and priests. Less well known is the use of muricid opercula in sacred incense and traditional medicines, although they are still used as rare ingredients today. This study provides the first chemical assessment of opercula from Muricidae, based on several traditional preparation procedures. Chemical analysis of opercula smoke revealed aromatic phenols, which act as fragrance stabilisers and produce a "medicinal" odour. Analysis of lipid extracts revealed pharmaceutically active compounds, including brominated indoles, choline esters and adenosine, consistent with their traditional medical applications. Depending on the preparation procedures, toxic pyridine was also detected. ICP-MS analysis of muricid opercula shows the presence of essential macro and microelements, as well as metals, some of which exceed the recommended safe levels for human use. Nevertheless, these findings support the Muricidae as an historically important marine resource, providing Biblical dyes, medicines and perfume. The opercula contains biologically active compounds and produces smoke containing volatile scent compounds, consistent with their identification as the most likely source of onycha, a controversial ingredient in sacred incense.

Marine molluscs are rich in biologically active natural products that provide new potential sources of anti-inflammatory agents. Here we used bioassay guided fractionation of extracts from the muricid Dicathais orbita to identify brominated indoles with anti-inflammatory activity, based on the inhibition of nitric oxide (NO) and tumour necrosis factor α (TNFα) in lipopolysaccharide (LPS) stimulated RAW264.7 macrophages and prostaglandin E2 (PGE2) in calcium ionophore-stimulated 3T3 ccl-92 fibroblasts. Muricid brominated indoles were then compared to a range of synthetic indoles to determine structure-activity relationships. Both hypobranchial gland and egg extracts inhibited the production of NO significantly with IC50 of 30.8 and 40 μg/mL, respectively. The hypobranchial gland extract also inhibited the production of TNFα and PGE2 with IC50 of 43.03 µg/mL and 34.24 µg/mL, respectively. The purified mono-brominated indole and isatin compounds showed significant inhibitory activity against NO, TNFα, and PGE2, and were more active than dimer indoles and non-brominated isatin. The position of the bromine atom on the isatin benzene ring significantly affected the activity, with 5Br > 6Br > 7Br. The mode of action for the active hypobranchial gland extract, 6-bromoindole, and 6-bromoisatin was further tested by the assessment of the translocation of nuclear factor kappa B (NFκB) in LPS-stimulated RAW264.7 mouse macrophage. The extract (40 µg/mL) significantly inhibited the translocation of NFκB in the LPS-stimulated RAW264.7 macrophages by 48.2%, whereas 40 µg/mL of 6-bromoindole and 6-bromoistain caused a 60.7% and 63.7% reduction in NFκB, respectively. These results identify simple brominated indoles as useful anti-inflammatory drug leads and support the development of extracts from the Australian muricid D. orbita, as a new potential natural remedy for the treatment of inflammation.

Muricidae are marine molluscs known for the production of Tyrian purple and bioactive precursor compounds. A validation study for the extraction and analysis of secondary metabolites found in the hypobranchial gland of the muricid Dicathais orbita is reported, using high performance liquid chromatography–mass spectrometry (HPLC-MS) with diode array detector (DAD). Quantification of the dominant secondary metabolites from D. orbita is described, followed by a comparison of solvent extraction procedures and stability studies. The intra- and inter-day relative standard deviation (RSD) for tyrindoxyl sulphate was 0.46% and 0.17%, respectively. The quantification was linear for standards murexine, 6-bromoisatin, and tyrindoxyl sulphate. The limits of detection were 0.03, 0.004, and 0.07 mg/mL, respectively, and the limits of quantification were 0.09, 0.01, and 0.22 mg/mL, respectively. The results showed that alcoholic solvents were better for extracting choline ester and indoxyl sulphate ultimate precursors, while chloroform was more suitable for the extraction of the intermediate precursors. Multivariate analysis revealed significant differences in extract composition according to the solvent used. Stability testing showed an increase of the oxidative compounds 6-bromoisatin and putative tyrindoxyl S-oxide sulphate in the ethanol extracts while more degradation products were seen in the chloroform extracts after months of cold storage. The validated method was found to be simple, reproducible, precise, and suitable for quantification of the secondary metabolites of muricid molluscs for dye precursor and nutraceutical quality control, as well as applications in marine chemical ecology.

The application of DNA barcoding to botanicals has attracted much attention recently, in part because this type of test has been used as the basis for high-profile regulatory actions.

Among the many DNA-based methods with potential application in the authentication of medicinal plant materials, DNA barcoding has attracted particular attention recently and has been touted by some as a superior authentication method. In order to assess the utility of DNA barcoding in the quality control of raw materials, extracts and finished herbal medicinal products, we examined four universal barcodes (matK, rbcL, trnH-psbA and ITS2) in 17 authentic raw materials and in extracts and tablets made from those raw materials. Authentic herbarium specimens were also included. Samples were chemically profiled and marker compounds identified by UPLC-MS. DNA was extracted with the NucleoSpin® 96 Plant II kit and each genetic target amplified by PCR, and Sanger sequenced. Results were compared to sequences in GenBank using BLAST. For raw materials, the rate of successful amplification and sequencing varied for the four targets but was low overall (highest for rbcL and trnH-psbA, 25% and 33% respectively). Identification to species was achieved for 18% (trnH-psbA), 15% (rbcL), 5% (matK) and 0% (ITS2) across all samples. Of dried materials, 53% were correctly identified by at least one barcode. Importantly, only one extract yielded amplifiable DNA; 20 extracts did not and could not be identified by any of the four barcodes. In contrast, the phytochemical integrity of all samples was confirmed by UPLC-MS.

Objective: Increased consumption of fruits and vegetables as functional foods leads to the reduction of signs of metabolic syndrome. The aim of this study was to measure and compare cardiovascular, liver, and metabolic parameters following chronic administration of the same dose of anthocyanins either from chokeberry (CB) or purple maize (PM) in rats with diet-induced metabolic syndrome.Methods: Male Wistar rats were fed a maize starch (C) or high-carbohydrate, high-fat diet (H) and divided into six groups for 16 wk. The rats were fed C, C with CB or PM for the last 8 wk (CCB or CPM), H, H with CB or PM for the last 8 wk (HCB or HPM); CB and PM rats received similar to 8 mg anthocyanins/kg daily. The rats were monitored for changes in blood pressure, cardiovascular and hepatic structure and function, glucose tolerance, and adipose tissue mass.Results: HCB and HPM rats showed reduced visceral adiposity index, total body fat mass, and systolic blood pressure; improved glucose tolerance, liver, and cardiovascular structure and function; decreased plasma triacylglycerols and total cholesterol compared with H rats. Inflammatory cell infiltration was reduced in heart and liver.Conclusion: CB and PM interventions gave similar responses, suggesting that anthocyanins are the bioactive molecules in the attenuation or reversal of metabolic syndrome by prevention of inflammation-induced damage. (C) 2016 Elsevier Inc. All rights reserved.

Both black (B) and green (G) cardamom are used as flavours during food preparation. This study investigated the responses to B and G in a diet-induced rat model of human metabolic syndrome. Male Wistar rats were fed either a corn starch-rich diet (C) or a high-carbohydrate, high-fat diet with increased simple sugars along with saturated and trans fats (H) for 16 weeks. H rats showed signs of metabolic syndrome leading to visceral obesity with hypertension, glucose intolerance, cardiovascular remodelling and nonalcoholic fatty liver disease. Food was supplemented with 3% dried B or G for the final eight weeks only. The major volatile components were the closely related terpenes, 1,8-cineole in B and α-terpinyl acetate in G. HB (high-carbohydrate, high-fat + black cardamom) rats showed marked reversal of diet-induced changes, with decreased visceral adiposity, total body fat mass, systolic blood pressure and plasma triglycerides, and structure and function of the heart and liver. In contrast, HG (high-carbohydrate, high-fat + green cardamom) rats increased visceral adiposity and total body fat mass, and increased heart and liver damage, without consistent improvement in the signs of metabolic syndrome. These results suggest that black cardamom is more effective in reversing the signs of metabolic syndrome than green cardamom.