Odor-active volatile compounds in preterm breastmilk.

BACKGROUND: Volatile compounds in breastmilk (BM) likely influence flavor learning and, through the cephalic phase response, metabolism, and digestion. Little is known about the volatile compounds present in preterm BM. We investigated whether maternal or infant characteristics are associated with the profile of volatile compounds in preterm BM. METHODS: Using solid-phase microextraction coupled with gas chromatography/mass spectrometry, we analyzed volatile compounds in 400 BM samples collected from 170 mothers of preterm infants. RESULTS: Forty volatile compounds were detected, mostly fatty acids and their esters (FA and FAe), volatile organic compounds (VOCs), aldehydes, terpenoids, alcohols, and ketones. The relative concentration of most FA and FAe increased with advancing lactation and were lower in BM of most socially deprived mothers and those with gestational diabetes (p < 0.05), but medium-chain FAs were higher in colostrum compared to transitional BM (p < 0.001). Infant sex, gestational age, and size at birth were not associated with the profile of volatile compounds in preterm BM. CONCLUSIONS: Sensory-active volatile FA and FAe are the major contributors to the smell of preterm BM. The associations between lactation stage, maternal characteristics, and volatile compounds, and whether differences in volatile compounds may affect feeding behavior or metabolism, requires further research. IMPACT: Sensory-active volatile FAs are major contributors to the smell of preterm BM and are influenced by the lactation stage and maternal characteristics. Longitudinal analysis of volatile compounds in preterm BM found that FAs increased with advancing lactation. Colostrum had a higher concentration of medium-chain FAs compared to transitional BM and the concentration of these is associated with socioeconomic status, gestational diabetes, and ethnicity.

Simultaneous Quantification of Organic Acids in Tamarillo (Solanum betaceum) and Untargeted Chemotyping Using Methyl Chloroformate Derivatisation and GC-MS.

Sixteen organic acids were quantified in peel and pulp of Amber, Laird's Large and Mulligan cultivars of tamarillo using GC-MS. Fourteen of these compounds had not previously been quantified in tamarillo. An untargeted metabolomics approach was used in parallel to identify and quantify 64 more metabolites relative to the internal standard, indicating abundances of glutamic acid, pro-line, aspartic acid and γ-aminobutyric acid as well as lower concentrations of several other essential fatty acids and amino acids. The main findings were that total organic acid concentration was significantly higher (p < 0.05) in pulp than in peel, with the highest concentration seen in Mulligan pulp (219.7 mg/g DW). Remarkably, after citric acid, the potent bactericide itaconic acid was the second most abundant organic acid. At least 95% of organic acids in tamarillo were one of these two acids, as well as cis-aconitic, malic and 4-toluic acids. Differences between cultivar chemotypes were as substantial as differences between tissues. These results suggest that the bitter flavour of the peel does not result from organic acids. The combination of targeted and untargeted metabolomics techniques for simultaneous qualitative and quantitative investigation of nutrients and flavours is efficient and informative.

Water relations determine short time leaf growth patterns in the mangrove Avicennia marina (Forssk.) Vierh.

High-resolution leaf growth is rarely studied despite its importance as a metric for plant performance and resource use efficiency. This is in part due to methodological challenges. Here, we present a method for in situ leaf growth measurements in a natural environment. We measured instantaneous leaf growth on a mature Avicennia marina subsp. australasica tree over several weeks. We measured leaf expansion by taking time-lapse images and analysing them using marker tracking software. A custom-made instrument was designed to enable long-term field studies. We detected a distinct diel growth pattern with leaf area shrinkage in the morning and leaf expansion in the afternoon and at night. On average, the observed daily shrinkage was 37% of the net growth. Most of the net growth occurred at night. Diel leaf area shrinkage and recovery continued after growth cessation. The amount of daily growth was negatively correlated with shrinkage, and instantaneous leaf growth and shrinkage were correlated with changes in leaf turgor. We conclude that, at least in this tree species, instantaneous leaf growth patterns are very strongly linked to, and most likely driven by, leaf water relations, suggesting decoupling of short-term growth patterns from carbon assimilation.

Profiling of headspace volatiles from Escherichia coli cultures using silicone-based sorptive media and thermal desorption GC-MS.

Headspace sorptive extraction technique using silicone based sorptive media coated stir bars is used for the first time here to extract, identify, and quantify heavy volatile organic compounds present in Escherichia coli culture headspace. Detection of infection presence is largely accomplished in laboratories through physical sampling and subsequent growth of cultures for biochemical testing. The use of volatile biomarkers released from pathogens as indicators for pathogenic presence can vastly reduce the time needed whilst improving the success rates for infection detection. To validate this, by using a contactless headspace sorptive extraction technique, the volatile compounds released from E. coli, grown in vitro, have been extracted and identified. Two different sorptive media for extracting these headspace volatiles were compared in this study and the identified volatiles were quantified. The large phase volume and wider retention of this sorptive technique compared to traditional sampling approach enabled preconcentration and collection of wider range of volatiles towards developing an extensive database of such heavy volatiles associated with E. coli. This supplements the existing data of potential bacterial markers and use of internal standards in these tests allows semi-quantitative estimation of these compounds towards the development and optimization of novel pathogen sensing devices.

Human Metabolism and Excretion of Kawakawa (Piper excelsum) Leaf Chemicals.

SCOPE: Piper excelsum (kawakawa) has a history of therapeutic use by Maori in Aotearoa New Zealand. It is currently widely consumed as a beverage and included as an ingredient in "functional" food product. Leaves contain compounds that are also found in a wide range of other spices, foods, and medicinal plants. This study investigates the human metabolism and excretion of kawakawa leaf chemicals. METHODS AND RESULTS: Six healthy male volunteers in one study (Bioavailability of Kawakawa Tea metabolites in human volunteers [BOKA-T]) and 30 volunteers (15 male and 15 female) in a second study (Impact of acute Kawakawa Tea ingestion on postprandial glucose metabolism in healthy human volunteers [TOAST]) consume a hot water infusion of dried kawakawa leaves (kawakawa tea [KT]). Untargeted Liquid Chromatography-Tandem Mass spectrometry (LC-MS/MS) analyses of urine samples from BOKA-T identified 26 urinary metabolites that are significantly associated with KT consumption, confirmed by the analysis of samples from the independent TOAST study. Seven of the 26 metabolites are also detected in plasma. Thirteen of the 26 urinary compounds are provisionally identified as metabolites of specific compounds in KT, eight metabolites are identified as being derived from specific compounds in KT but without resolution of chemical structure, and five are of unknown origin. CONCLUSIONS: Several kawakawa compounds that are also widely found in other plants are bioavailable and are modified by phase 1 and 2 metabolism.

4-Ethylphenol-fluxes, metabolism and excretion of a gut microbiome derived neuromodulator implicated in autism.

Gut-microbiome-derived metabolites, such as 4-Ethylphenol [4EP], have been shown to modulate neurological health and function. Although the source of such metabolites is becoming better understood, knowledge gaps remain as to the mechanisms by which they enter host circulation, how they are transported in the body, how they are metabolised and excreted, and the way they exert their effects. High blood concentrations of host-modified 4EP, 4-ethylphenol sulfate [4EPS], are associated with an anxiety phenotype in autistic individuals. We have reviewed the existing literature and discuss mechanisms that are proposed to contribute influx from the gut microbiome, metabolism, and excretion of 4EP. We note that increased intestinal permeability is common in autistic individuals, potentially explaining increased flux of 4EP and/or 4EPS across the gut epithelium and the Blood Brain Barrier [BBB]. Similarly, kidney dysfunction, another complication observed in autistic individuals, impacts clearance of 4EP and its derivatives from circulation. Evidence indicates that accumulation of 4EPS in the brain of mice affects connectivity between subregions, particularly those linked to anxiety. However, we found no data on the presence or quantity of 4EP and/or 4EPS in human brains, irrespective of neurological status, likely due to challenges sampling this organ. We argue that the penetrative ability of 4EP is dependent on its form at the BBB and its physicochemical similarity to endogenous metabolites with dedicated active transport mechanisms across the BBB. We conclude that future research should focus on physical (e.g., ingestion of sorbents) or metabolic mechanisms (e.g., conversion to 4EP-glucuronide) that are capable of being used as interventions to reduce the flux of 4EP from the gut into the body, increase the efflux of 4EP and/or 4EPS from the brain, or increase excretion from the kidneys as a means of addressing the neurological impacts of 4EP.

Acute Effects of Kawakawa (Piper excelsum) Intake on Postprandial Glycemic and Insulinaemic Response in a Healthy Population.

Background: Piper excelsum (kawakawa) is an endemic shrub of Aotearoa, New Zealand, of cultural and medicinal importance to Maori. Its fruits and leaves are often consumed. These tissues contain several compounds that have been shown to be biologically active and which may underpin its putative health-promoting effects. The current study investigates whether kawakawa tea can modulate postprandial glucose metabolism. Methods: We report a pilot three-arm randomized crossover study to assess the bioavailability of kawakawa tea (BOKA-T) in six male participants with each arm having an acute intervention of kawakawa tea (4 g/250 mL water; 1 g/250 mL water; water) and a follow-up two-arm randomized crossover study to assess the impact of acute kawakawa tea ingestion on postprandial glucose metabolism in healthy human volunteers (TOAST) (4 g/250 mL water; and water; n = 30 (15 male and 15 female)). Participants consumed 250 mL of kawakawa tea or water control within each study prior to consuming a high-glycemic breakfast. Pre- and postprandial plasma glucose and insulin concentrations were measured, and the Matsuda index was calculated to measure insulin sensitivity. Results: In the BOKA-T study, lower plasma glucose (p < 0.01) and insulin (p < 0.01) concentrations at 60 min were observed after consumption of a high-dose kawakawa tea in comparison to low-dose or water. In the TOAST study, only plasma insulin (p = 0.01) was lower at 60 min in the high-dose kawakawa group compared to the control group. Both studies showed a trend towards higher insulin sensitivity in the high-dose kawakawa group compared to water only. Conclusions: Consuming kawakawa tea may modulate postprandial glucose metabolism. Further investigations with a longer-term intervention study are warranted.

Exploring the Chemical Space of Kawakawa Leaf (Piper excelsum).

The chemical profiles of kawakawa (Piper excelsum) leaves were analysed through targeted and non-targeted LC-MS/MS. The phytochemical profile was obtained for both aqueous extracts representative of kawakawa tea and methanolic extracts. Sixty-four compounds were identified from eight leaf sources including phenylpropanoids, lignans, flavonoids, alkaloids and amides. Eight of these compounds were absolutely quantified. The chemical content varied significantly by leaf source, with two commercially available sources of dried kawakawa leaves being relatively high in phenylpropanoids and flavonoids compared with field-collected fresh samples that were richer in amides, alkaloids and lignans. The concentrations of pharmacologically active metabolites ingested from the traditional consumption of kawakawa leaf as an aqueous infusion, or from novel use as a seasoning, are well below documented toxicity thresholds.

Fat-Soluble Vitamers: Parent-Child Concordance and Population Epidemiology in the Longitudinal Study of Australian Children.

Fat-soluble vitamers (FSV) are a class of diverse organic substances important in a wide range of biological processes, including immune function, vision, bone health, and coagulation. Profiling FSV in parents and children enables insights into gene-environment contributions to their circulating levels, but no studies have reported on the population epidemiology of FSV in these groups as of yet. In this study, we report distributions of FSV, their parent-child concordance and variation by key characteristics for 2490 children (aged 11-12 years) and adults (aged 28-71 years) in the Child Health CheckPoint of the Longitudinal Study of Australian Children. Ten A, D, E and K vitamers were quantified using a novel automated LC-MS/MS method. All three K vitamers (i.e., K1, MK-4, MK-7) and 1-α-25(OH)2D3 were below the instrument detection limit and were removed from the present analysis. We observed a strong vitamer-specific parent-child concordance for the six quantifiable A, D and E FSVs. FSV concentrations all varied by age, BMI, and sex. We provide the first cross-sectional population values for multiple FSV. Future studies could examine relative genetic vs. environmental determinants of FSV, how FSV values change longitudinally, and how they contribute to future health and disease.

Cortical Oxygenation Changes during Gastric Tube Feeding in Moderate- and Late-Preterm Babies: A NIRS Study.

Smell and taste of food can trigger physiological responses facilitating digestion and metabolism of nutrients. Controlled experimental studies in preterm babies have demonstrated that smell activates the orbitofrontal cortex (OFC) but none have investigated the effect of taste stimulation. Using cotside Near-Infrared Spectroscopy (NIRS), we measured changes in OFC cerebral oxygenation in response to gastric tube feeds five and 10 days after birth in 53 assessments of 35 moderate- to late-preterm babies enrolled in a randomized trial. Babies were randomly assigned to receive smell and taste of milk before gastric tube feeds (intervention group, n = 16) or no exposure (control group, n = 19). The majority of babies were born at 33 weeks of gestation (range 32-34) and 69% were boys. No differences in OFC cerebral oxygenation were observed between control and intervention groups. Gastric tube feeds induced activation of the OFC (p < 0.05), but sensory stimulation alone with smell and taste did not. Boys, but not girls, showed activation of the OFC following exposure to smell of milk (p = 0.01). The clinical impact of sensory stimulation prior to tube feeds on nutrition of preterm babies, as well as the impact of environmental inputs on cortical activation, remains to be determined.

Volatile Components and Preliminary Antibacterial Activity of Tamarillo (Solanum betaceum Cav.).

Tamarillo is a nutrient-dense fruit with a unique aroma from its volatile compounds (VCs). In this study, we aimed to compare the volatile profiles: (i) of fresh and freeze-dried tamarillo; (ii) detected using Thermal Desorption-Gas Chromatography-Mass Spectrometry (TD-GC-MS) and Solid-Phase MicroExtraction-Gas Chromatography-Mass Spectrometry (SPME-GC-MS); (iii) of freeze-dried pulp and peel of New Zealand grown tamarillo. The possible antibacterial activity of freeze-dried tamarillo extracts was also investigated. We show that freeze-drying maintained most of the VCs, with some being more concentrated with the loss of water. The most abundant VC in both fresh and freeze-dried tamarillo was hexanoic acid methyl ester for pulp (30% and 37%, respectively), and (E)-3-Hexen-1-ol for peel (36% and 29%, respectively). With the use of TD-GC-MS, 82 VCs were detected for the first time, when compared to SPME-GC-MS. Methional was the main contributor to the overall aroma in both peel (15.4 ± 4.2 µg/g DW) and pulp (118 ± 8.1 µg/g DW). Compared to water as the control, tamarillo extracts prepared by water and methanol extraction showed significant antibacterial activity against E. coli, P. aeruginosa, and S. aureus with zone of inhibition of at least 13.5 mm. These results suggest that freeze-dried tamarillo has a potential for use as a natural preservative to enhance aroma and shelf life of food products.

Phenolic and Anthocyanin Compounds and Antioxidant Activity of Tamarillo (Solanum betaceum Cav.).

This study examined phenolics and anthocyanins present in Amber, Laird's Large and Mulligan cultivars of tamarillo that were cultivated in -Whangarei, Northland of New Zealand. Samples were further separated by their tissue types, peel and pulp. Using LC-MS/MS, twelve polyphenols were quantified and six (ellagic acid, rutin, catechin, epicatechin, kaempferol-3-rutinoside and isorhamnetin-3-rutinoside) were detected for the first time in tamarillo. Mulligan cultivar showed the highest amounts of phenolic and anthocyanin compounds and the highest antioxidant activity. Phenolic compounds were mostly synthesized from shikimic acid route, and chlorogenic acid dominated the profile regardless of cultivar and tissue types. Anthocyanin profile was dominated by delphinidin-3-rutinoside in pulp. Higher amounts of anthocyanins were detected in this study, which may be explained by favourable growth conditions (high light intensity and low temperature) for anthocyanin biosynthesis in New Zealand. Higher antioxidant activity and total phenolic content in peels than in pulps were found when assessed by Cupric Ion-Reducing Antioxidant Capacity (CUPRAC), Ferric Reducing Ability of Plasma (FRAP) and Folin-Ciocalteu assays, and a positive correlation (-r > 0.9, p ≤ 0.01) between the three assays was observed. Current findings endorse that tamarillo has a great bioactive potential to be developed further as a functional ingredient with considerable levels of antioxidant compounds and antioxidant activity.

Olfactory Cues in Infant Feeds: Volatile Profiles of Different Milks Fed to Preterm Infants.

Background: Smell is determined by odor-active volatile compounds that bind to specific olfactory receptors, allowing us to discriminate different smells. Olfactory stimulation may assist with digestion and metabolism of feeds in the neonate by activation of the cephalic phase response of digestion. Infants' physiological responses to the smell of different milks suggest they can distinguish between breastmilk and infant formula. We aimed to describe the profile of volatile compounds in preterm breastmilk and investigate how this differed from that of other preterm infant feeding options including pasteurized donor breastmilk, breastmilk with bovine milk-based fortifier, human milk-based products and various infant formulas. Methods: Forty-seven milk samples (13 different infant formulas and 34 human milk-based samples) were analyzed. Volatile compounds were extracted using Solid Phase Micro Extraction. Identification and relative quantification were carried out by Gas Chromatography with Mass Spectrometry. Principal Component Analysis (PCA) and one-way Analysis of Variance (ANOVA) with Tukey's HSD (parametric data) or Conover's post-hoc test (non-parametric data) were used as appropriate to explore differences in volatile profiles among milk types. Results: In total, 122 compounds were identified. Breastmilk containing bovine milk-based fortifier presented the highest number of compounds (109) and liquid formula the lowest (70). The profile of volatile compounds varied with 51 compounds significantly different (adjusted p < 0.001) among milk types. PCA explained 47% of variability. Compared to preterm breastmilk, the profile of volatile compounds in breastmilk with added bovine milk-based fortifier was marked by presence of fatty acids and their esters, ketones and aldehydes; infant formulas were characterized by alkyls, aldehydes and furans, and human milk-based products presented high concentrations of aromatic hydrocarbons, terpenoids and specific fatty acids. Conclusions: Sensory-active products of fatty acid oxidation are the major contributors to olfactory cues in infant feeds. Analysis of volatile compounds might be useful for monitoring quality of milk and detection of oxidation products and environmental contaminants. Further research is needed to determine whether these different volatile compounds have biological or physiological effects in nutrition of preterm infants.

Quantification of Carotenoids, α-Tocopherol, and Ascorbic Acid in Amber, Mulligan, and Laird's Large Cultivars of New Zealand Tamarillos (Solanum betaceum Cav.).

Amber (yellow), Laird's Large (red) and Mulligan (purple-red) cultivars of New Zealand tamarillo fruit were separated into pulp (endo- and mesocarp) and peel (exocarp), and analyzed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) for carotenoids, α-tocopherol and ascorbic acid contents. Fresh Mulligan pulp had the highest content of ß-carotene (0.9 mg/100 g), α-tocopherol (1.9 mg/100 g), and ascorbic acid (28 mg/100 g). Higher concentrations of ß-carotene and ascorbic acid, and lower concentrations of α-tocopherol were detected in pulps compared with peels. Compared with standard serves of other fruit, tamarillo had the highest ß-carotene (9-20% RDI (recommended dietary intake)/serve), high ascorbic acid (67-75% RDI/serve), and α-tocopherol (16-23% adequate intake/serve). All cultivars had diverse carotenoid profiles dominated by provitamin A carotenoids (ß-carotene and ß-cryptoxanthin) and xanthophyll carotenoids (lutein; zeaxanthin and antheraxanthin). Favorable growth conditions (high light intensity and low temperature) may explain the higher antioxidant vitamin content in New Zealand tamarillos compared to those from other countries. Tamarillo peels may be used as natural food coloring agent to reduce waste and deliver sustainable production.

Validation and application of a modified QuEChERS method for extracting neonicotinoid residues from New Zealand maize field soil reveals their persistence at nominally hazardous concentrations.

The widespread use of neonicotinoid insecticides is controversial due to their persistence in the environment and concern for the long-term consequences of their use. We present a simple, low-cost method for the sensitive and efficient extraction from soil of thiamethoxam, clothianidin and imidacloprid, the three neonicotinoid insecticides approved in New Zealand as seed coatings. We have validated this method by applying it to uncontaminated soil samples spiked with environmentally relevant concentrations of the three targets. Absolute recoveries were >80% with instrument detection limits <1 ng g-1 wet soil. We also applied the method to soil samples collected from maize fields in New Zealand's North Island and found imidacloprid in 43 out of 45 samples and clothianidin in every one. Mean imidacloprid concentrations varied from 0.5 to 9.4 ng g-1 (wet weight) and clothianidin from 2.1 to 26.7 ng g-1 (wet weight). Imidacloprid concentrations exceed the New Zealand Environmental Protection Agency's Environmental Exposure Limit of 1 ng g-1 (dry weight) at eight of the nine sites sampled. These results are remarkable because we have detected multiple neonicotinoid residues at every site sampled. Imidacloprid residues appear to persist at significant concentrations at five of our sites from an application at least two years previously. This is only the third study to report the presence of neonicotinoid residues in NZ's environment and the first to show that those residues are persistent in the environment at nominally hazardous concentrations.

Metal composition of arrow squid (Nototodarus sloanii [Gray 1849]) from the Chatham Rise, New Zealand: implications for human consumption.

Cephalopods are important in the diets of humans and many other apex predators, and can play an important role in the bioaccumulation of metals. In this study, metal concentrations were analysed in the commercially and ecologically important southern arrow squid, Nototodarus sloanii (Gray 1849), from a heavily targeted fisheries area on New Zealand's Chatham Rise. A variety of tissue types were compared in order to assess the bioaccumulation in edible tissues (mantle and arms) and other organs (digestive gland, kidney, and hearts). Although metal concentrations varied among tissue types, the highest concentrations were found in the digestive gland (for Cd, Cu, Fe, Ni, Zn) and branchial hearts (for Cr, U). We report the first Cd concentrations for N. sloanii from the Chatham Rise, with mean values of 3.11 µg. g-1 in the mantle and 102.53 µg. g-1 in the digestive gland. Our data suggest that concentrations observed in the mantle tissue (which forms the majority of the muscle tissue) and digestive gland (the primary organ for metal accumulation) can be used to estimate the animal's total body burden for all metals analysed. The toxicological and dietary consequences for predators (including humans) feeding on arrow squid can be inferred from measurements of mass and metal concentration in these tissues. Arrow squid represent an important vector for Cd transfer within the pelagic Chatham Rise food web. These are the first recorded baseline data for metal concentrations for any squid in this oceanic region.

Red cherries (Prunus avium var. Stella) processed by pulsed electric field - Physical, chemical and microbiological analyses.

This study examined, for the first time, the effect of mild or moderate intensity pulsed electric field (PEF) processing on cherries, in particular changes in physicochemical properties, release of anthocyanins and polyphenols, and the potential growth of lactic acid bacteria. Cherry samples were treated at a constant pulse frequency of 100Hz and a constant pulse width of 20µs with different electric field strengths between 0.3 and 2.5kV/cm. Titratable acidity and total soluble solids values of most PEF samples stored for 24h significantly decreased compared to other samples. Stored samples also had increased cyanidin glucoside content. However, concentration of rutin, 4-hydroxybenzoic acid and isorhamnetin rutinoside significantly decreased in samples stored for 24h. In conclusion, sweet cherries were only influenced by storage after PEF processing. PEF processing did not affect the growth of probiotic bacteria.

Offshore iron sand extraction in New Zealand: Potential trace metal exposure of benthic and pelagic biota.

Plans to exploit an offshore source of iron sand in South Taranaki Bight (STB), New Zealand, caused concerns that such exploitation may expose benthic and pelagic biota to elevated trace metal concentrations. We conducted dilute-acid extractions and standard elutriate tests to investigate the potential of this exploitation to (1) create a new seafloor with elevated trace metal content, (2) mobilise trace metals during iron sand extraction and, (3) enrich the returning process seawater, which feeds iron sand through mills, with trace metals. We found that recruits of freshly uncovered sediment may encounter higher-than-natural concentrations of cadmium, nickel and chromium (but not of copper, lead, and zinc) and propose to investigate the bioavailability of these metals. Elutriate test with raw and milled iron sand revealed that, for nickel and copper, dilution of the process seawater may be required to meet the local water quality guideline. We argue that this dilution can be achieved by adjustment of the mass and seawater balance of the offshore extraction process.

Integrated assessment of oil pollution using biological monitoring and chemical fingerprinting.

A full assessment of the impact of oil and chemical spills at sea requires the identification of both the polluting chemicals and the biological effects they cause. Here, a combination of chemical fingerprinting of surface oils, tissue residue analysis, and biological effects measures was used to explore the relationship between spilled oil and biological impact following the grounding of the MSC Napoli container ship in Lyme Bay, England in January 2007. Initially, oil contamination remained restricted to a surface slick in the vicinity of the wreck, and there was no chemical evidence to link biological impairment of animals (the common limpet, Patella vulgata) on the shore adjacent to the oil spill. Secondary oil contamination associated with salvage activities in July 2007 was also assessed. Chemical analyses of aliphatic hydrocarbons and terpanes in shell swabs taken from limpet shells provided an unequivocal match with the fuel oil carried by the ship. Corresponding chemical analysis of limpet tissues revealed increased concentrations of polycyclic aromatic hydrocarbons (PAHs) dominated by phenanthrene and C1 to C3 phenanthrenes with smaller contributions from heavier molecular weight PAHs. Concurrent ecotoxicological tests indicated impairment of cellular viability (p < 0.001), reduced immune function (p < 0.001), and damage to DNA (Comet assay, p < 0.001) in these animals, whereas antioxidant defenses were elevated relative to un-oiled animals. These results illustrate the value of combining biological monitoring with chemical fingerprinting for the rapid identification of spilled oils and their sublethal impacts on biota in situ.

The metabolic and fitness costs associated with metal resistance in Nereis diversicolor.

The population of Nereis diversicolor inhabiting the upper reaches of Restronguet Creek, Cornwall, UK is highly resistant to acute zinc and copper toxicity. Here we employ bioenergetic accounting and fecundity counts to demonstrate the energetic costs associated with this phenomenon in terms of the worms' allocation of metabolic resources and reproductive output [P(r)]. Metal-resistant animals exhibited a scope for growth that was 46-62% less than that of animals from two non-resistant reference populations, corresponding to a mean metabolic cost of 1.31 mJ h(-1)mg DW(-1). The resistant population also contained 13% less lipid than animals from the reference populations and 73-81% less carbohydrates. Consequently, mass-specific fecundity was reduced in resistant animals by 39-45%, although material investment in individual gametes did not appear to vary. This demonstrates fitness costs associated with metal resistance in this ecologically important polychaete and adds to our understanding of phenotypic trade-offs associated with resistance.